Peptide Dendrimers Aplication and Synthesis

Reviews in Molecular Biotechnology 90 2002.
195 229
Peptide dendrimers: applications and synthesis

Kristen Sadler, James P. TamU
Department of Microbiology and Immunology, Vanderbilt Uni ersity, A5119 MCN, Nash ille, TN 37232, USA
Abstract Peptide dendrimers are radial or wedge-like branched macromolecules consisting of a peptidyl branching core andror covalently attached surface functional units. The multimeric nature of these constructs, the unambiguous composition and ease of production make this type of dendrimer well suited to various biotechnological and biochemical applications. Applications include use as biomedical diagnostic reagents, protein mimetics, anticancer and antiviral agents, vaccines and drug and gene delivery vehicles. This review focuses on the different types of peptide dendrimers currently in use and the synthetic methods commonly employed to generate peptide dendrimers ranging from stepwise solid-phase synthesis to chemoselective and orthogonal ligation. 2002 Elsevier Science B.V. All rights reserved.
Keywords: Review; Peptide dendrimer; Multiple antigenic peptide; MAP; Ligation chemistry
1. Introduction Dendrimers are polymers with three distinct structural features: a central core; surface functionalities; and branching units that link the two. Peptide dendrimers can be broadly dened as any dendrimer that contains peptide bonds. This definition would, in theory, include a dendrimer with an amino acid core, branching units, surface functional groups or any combination of the three as a peptide dendrimer. This denition could be furU
Corresponding author. Tel.: q1-615-343-1465; fax: q1615-343-1467. E-mail address: [email protected] J.P. Tam..
ther broadened if we consider all types of amino acids including naturally occurring -amino acids as well as unnatural amino acids that have been utilized in peptide dendrimer synthesis as both branching units and surface functional groups. For example, unnatural amino acids such as amino acids have also been used as branching units Esfand and Tomalia, 2001.. In practice, the majority of peptide dendrimers being referred to in the literature and currently in use are based on the multiple antigen peptide MAP. system that consists of only two of the three structural features: branching units and surface functional groups. Interestingly, these molecules contain both -peptide and -peptide Tam, 1988, 1996.. In addition, they are often synthesized without a
1389-0352r02r$ - see front matter 2002 Elsevier Science B.V. All rights reserved. PII: S 1 3 8 9 - 0 3 5 2 0 1 . 0 0 0 6 1 - 7
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K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229 Table 1 Current and potential applications of polyvalent dendrimers Vehicles for delivery of Nucleic acids Encapsulated drugs Covalently linked drugs, e.g. anticancer agents Diagnostic reagents in Serodiagnosis Biosensor systems Magnetic resonance imaging Vaccines against Bacteria Virus Parasites Inhibitors of Pathogenic infections Inammatory response Autoimmune disease Cancer metastasis Modication of Cell cell interactions Gene expression, e.g. alteration of transcription factors binding to DNA
core and more appropriately termed dendrons rather than classical dendrimers. However, for convenience we will refer to all branched polypeptide constructs as dendrimers.
2. General characteristics of peptide dendrimers By virtue of their dendritic architecture, peptide dendrimers are frequently utilized as protein and liposomal mimetics as well as biomaterials in the life sciences and biomedical applications. Dendrimer design exploits two traits that are frequently observed in many naturally occurring systems: globular structure and polyvalency. Polyvalent interactions occur through the association of two or more ligands and receptors of the same type. Multivalent molecules are commonly found in nature ranging from small molecules of polysaccharides, nucleic acids and peptides to protein aggregates on virions, bacteria and other cells that aid in adhesion and cell cell interactions. The properties of multiple simultaneous interactions can differ markedly from those displayed by the constituents during monovalent interactions, suggesting that dendrimers with multiple functional sites can be used in the design of new drugs and research reagents. Such strategies to cluster ligands and functional groups on dendrimeric structures are attracting considerable attention in different areas from the production of biomaterials to vaccine technology. Dendrimeric materials are nding applications as antiviral agents Zanini and Roy, 1998., vehicles for delivery of nucleic acids Haensler and Szoka, 1993; Kukowska-Latallo et al., 1996; DeLong et al., 1997. and drugs Esfand and Tomalia, 2001., as biomedical tools such as magnetic resonance imaging contrast agents Kim and Zimmerman, 1998., protein mimetics, vaccines directed towards bacterial, viral and parasitic pathogens, as well as diagnostic reagents and molecular inhibitors. In the rst step of infection most pathogens such as bacteria and virus particles adhere to potential host cells via polyvalent interactions Mammen et al., 1998.. Therefore, there may be a role for multivalent dendrimers capable of mimicking receptors and ligands in preventing
infections. Similarly, cell cell interactions that are mediated by specic receptor ligand associations can be studied in ne detail through the use of dendrimers. Pathophysiological responses such as inammation and autoimmune reactions could be prevented by therapeutic use of dendrimers. Some of the current and potential applications of synthetic dendrimers in biomedicine and biotechnology are outlined in Table 1. Peptide dendrimers vary from low molecular weight species of 2 kDa to large protein-like constructs ) 100 kDa. The size and complexity of the individual dendrimers are determined by two factors, the number of layers of branching units often referred to as the generation number. and the surface supporting the terminal functional groups which can be large peptides or proteins of substantial size. Typically, peptide dendrimers have generation numbers between 2 and 32. Similar to other dendrimers, synthesis of peptide dendrimers is tightly controlled with products of consistent size, architecture and composition.
K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229
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Peptide dendrimers can be divided into three types. The rst are grafted peptide dendrimers. These are conventional dendrimers with either unnatural amino acids or organic groups as the branching core and peptide or proteins attached as surface functional groups. Of the three, the grafted peptide dendrimer is the largest in terms of size because they generally contain high generation numbers of branching cores. In contrast, peptide dendrimers of the second type are essentially branching polyamino acids. Consequently, they tend to be the smallest by size with the core consisting of natural amino acids and the terminal amino acids acting as surface functional groups. The third type consisting of mostly peptides has been traditionally known as peptide dendrimers. In this group, with MAPs as the most well known example, the core consists of amino acids and the surface functional groups are also peptidyl chains. This review provides some details on the rst two types of peptide dendrimers, however, we will largely focus on the third type because this type is used most frequently in biological and biochemical applications. 2.1. Grafted peptide dendrimers with unnatural amino acids and organic cores Peptide dendrimers based on the commercially available poly amidoamine., or PAMAM core may be considered classical dendrimers and one of the most widely used Esfand and Tomalia, 2001.. They are synthesized by a divergent method involving a two-step iterative reaction sequence that produces concentric shells of dendritic alanine units. Various functional groups, including peptides, have been covalently linked to the PAMAM dendrimer and used for different applications. Anticancer drugs may be encapsulated by PAMAM dendrimers with polyethylene glycol. grafts prior to delivery in order to increase efcacy Kojima et al., 2000.. Addition of sugars such as lactose and fructose to the PAMAM dendrimer produces glycodendrimers that are potential high-afnity ligands for sugar receptors Andre et al., 1999; Kawase et al., 2000.. Excellent reviews by Esfand and Tomalia 2001., Kim and Zimmerman 1998. outline the potential
biomedical applications of PAMAM dendrimers with carbohydrate and nucleic acid functional units. Peptides have also been attached to the PAMAM core. In one of the most promising studies an articial photosynthetic system has been constructed with amphipathic -helix peptides acting as surface functional groups attached to the core through a thioether linkage Sakamoto et al., 2001a,b.. The PAMAM dendrimer has also been used to generate a biochemical reagent Singh, 1998. by reacting a PAMAM core with calf intestine alkaline phosphatase and a Fab fragment of an anti-creatine kinase MB isoenzyme antibody. 2.2. Peptide dendrimers with polyamino acids as branching units and as surface functional sites Water-soluble amphipathic peptide dendrimers comprised only of three or four generations of lysine residues have been constructed to aid in the delivery of DNA to intact cells. Dendrimers consisting of a branched lysine core and 8 or 16 terminal surface amines may be mixed in solution with plasm id D NA and left to form dendrimer DNA complexes. The complexes are subsequently taken up into cells and the gene products expressed with high efciency compared to cells exposed to DNA alone Choi et al., 1999; Toth et al., 1999; Shah et al., 2000.. In a similar vein, lipidated peptide dendrimers consisting of polyamide groups 4, 8 or 16 terminal lysines. were linked through the rst generation lysine to three alkyl chains C 14 . and their interaction with charged and neutral liposomes was studied Purohit et al., 2001.. It was found that the interaction efciency of the dendrimers was greater than 88% and that the amount of dendrimer adsorbed decreased as the dendrimer size increased. The mechanism of interaction is unclear but adsorption is most likely due to hydrophobic interactions rather than electrostatic because the dendrimers interacted with equal efciency with negatively and positively charged liposomes. Lipidated peptide dendrimers have also been synthesized by the addition of 2-amino tetradecanoic acid to a polylysine support to create a dendrimer with 16 surface alkyl chains Florence et al., 2000..
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This peptide dendrimer was used to study the uptake of lipidated molecules by the mucosal surfaces of the digestive tract in mice. 2.3. Peptide dendrimers with amino acid branching units and surface peptidyl chains Peptide dendrimers comprised of amino acid cores and surface peptide chains Fig. 1. have been used most extensively in biological applications. The underlying design principle is that the polyvalency of the peptide dendrimers enhances the afnity of peptide-specic interactions with peptides, proteins and carbohydrates, thus enabling the study of receptor ligand interactions, the introduction of increased binding afnities and also development of new techniques for afnity purication and diagnostic tests. An example showing the ability of multimeric peptides to enhance binding afnity was described by Fassina et al. 1992a. where analytical HPLC and solid phase binding assays were used to evaluate the binding interactions between two complementary peptides in monomeric and octameric forms. Binding afnity was found to increase when one or both of the sequences were synthesized as octameric dendrimers. The interaction between the multimeric peptides was of similar afnity to that of polyclonal antibody and multimeric peptide illustrating that weak interactions may be magnied through the use of peptide dendrimers allowing more complete analysis. This may be potentially useful for characterizing the weak interactions between cell-surface receptors and ligands. Some of the biochemical applications of peptide dendrimers are listed in Table 2.
Fig. 1. Schematic illustration of peptide dendrimers that vary in size.
3. Peptide dendrimers in immunoassays and serodiagnosis Many immunoassays rely on the attachment of antigens to a solid surface such as plastic microtitre wells and their detection by addition of specic reagents. Short synthetic peptides readily available through chemical synthesis are generally considered to be ideal antigens because of the high specicity they confer. Unfortunately, most
short peptides are poor antigens due to their inability to bind to solid surfaces. The multimeric nature of peptide dendrimers has been found to aid in overcoming such deciencies by providing increased surface-binding character and sensitivity of detection Tam and Zavala, 1989; Marguerite et al., 1992; Briand et al., 1992; Marsden et al., 1992.. Adesida et al. 1999. exploited the increased afnity of protein interaction with peptide dendrimers compared to monomeric peptides in a study focusing on the interaction of immunoglobulins with a non-epitopic peptide in both direct and indirect peptide-antibody binding assays. In serodiagnosis where serum samples from naturally immunized or infected individuals are tested for the presence of specic antibody, the multimeric arrangement of dendrimers may also improve the detection of low afnity antibodies during early stages of infection, particularly those of the IgM isotype Habluetzel et al., 1991; Marsden et al., 1992.. In a dramatic example, a dendrimer shows a sensitivity increase of over 10 8-fold when compared to a peptide antigen in an enzyme-linked immunosorbent assay. Immunosorbent assays may be superceded in the near future by biosensor technology where dendrimers are bound directly to sensory chips and the strength of antibody interaction monitored with great accuracy Gomara et al., 2000a.. The use of dendrimers in biosensor technology requires further
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examination, particularly to optimize immobilization and ensure that factors such as steric hindrance Francis et al., 1991; Le et al., 1998. and interchain aggregation do not impede immobilization nor antibody recognition. The introduction
of spacer residues such as glycine between the lysine core and peptide sequence of interest Marsden et al., 1992. may reduce inhibitory interchain aggregation. The dendrimers must also be bound to the sensory chips in the correct
Table 2 Diagnostic and biochemical uses of peptide dendrimers Application Immunoassays and serodiagnosis Malaria Cirrhosis HIV-1 References
Tam and Zavala 1989., Habluetzel et al. 1991. Avila et al. 2001. Briand et al. 1992. Marsden et al. 1992., Estaquier et al. 1993. Vogel et al. 1994., Shin et al. 1997. Marguerite et al. 1992. Sabbatini et al. 1993a., Caponi et al. 1995. Marchini et al. 1994. Jackwood and Hilt 1995. Gomara et al. 2000a,b. Adesida et al. 1999. Tres and Kierszenbaum 1996.
Schistosoma mansoni Systemic lupus erythematosus Epstein-Barr virus Infectious bronchitis virus Hepatitis A virus Non-epitopic peptide Sperm antigen Epitope mapping and ligand binding Bluetongue virus Systemic lupus erythematosus Hepatitis C virus P. falciparum Pf72rHsp70-1 antigen NKTag, tumour antigen Inhibitors Macroautophagia and proteolysis Tumour growth and metastasis Enzyme inhibitors HIV-1 fusion and infection Interleukin 6 Sporozoite, malaria Fibronectin Antagonist of C5a Articial proteins Minicollagen Synthetic enzyme Biochemical studies Afnity purication of antibodies Presentation of T-cell epitopes Afnity purications
Yang et al. 1992. Sabbatini et al. 1993b. Simmonds et al. 1993. Dat et al. 2000. Jaso-Friedmann et al. 1996.
Miotto et al. 1994., Mortimore et al. 1994. Nomizu et al. 1993., Kim et al. 1994. Iwamoto et al. 1996., Manki et al. 1998. Fassina and Cassani 1993. Yahi et al. 1994a,b, 1995., Weeks et al. 1994. Wallace et al. 1994. Sinnis et al. 1994. Ingham et al. 1994. Kaneko et al. 1995.
Fields et al. 1993. Hahn et al. 1990.
Butz et al. 1994., Yang et al. 1999. Grillot et al. 1993. Fassina 1992., Fassina et al. 1992a,b. Yao et al. 1994. Sheldon et al. 1995.
Intracellular delivery
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orientation, for this ligation reactions may be exploited for example thiol exchange, hydrazide or sulfhydryl group coupling Schuck, 1997..
enhanced and a potentially useful therapeutic agent produced.
5. Peptide dendrimers as mimetics 4. Peptide dendrimers as inhibitors Another promising application of peptide dendrimers has been realized in the general design of inhibitors. Multimerization of binding elements such as charge in a dendrimer is controllable and provides an unambiguous structure that can be rened by analogs with relative ease. The inhibition of entry of malaria parasites into hepatocytes Sinnis et al., 1994. is a ne example of this application. Sinnis et al. 1994. have taken advantage of the fact that a cationic conserved sequence of malaria circumsporozoite CS. protein known as region II-plus requires aggregation for binding to liver hepatocyte proteoglycans. A tetramer containing region II-plus was synthesized and subsequently found to inhibit CS protein binding to hepatocytes in mice. A peptide dendrimer based on an eight-residue sequence from the third variable region V3 loop. of gp120, the HIV-1 surface envelope glycoprotein, has been shown to inhibit HIV-1 infection of both CD4q and CD4y cells Yahi et al., 1995; Carlier et al., 2000.. The increased binding afnity of dendrimers compared to monomeric peptides is becoming an important factor in the design of peptides aimed at inhibiting metastasis of various types of cancer cells. One of the most advanced studies has shown that a dendrimer containing 16 copies of a peptide from laminin a protein of the basement membrane extracellular matrix. signicantly inhibited tumor growth in vivo Nomizu et al., 1993.. This inhibitory activity increased as the peptide copy number increased, i.e. 16 ) 8 ) 4. The enhanced anti-tumor effects of the 16-mer were subsequently partly attributed to the ability of the multimer to compete more successfully with laminin for the laminin receptor on tumor cells thereby blocking the binding of malignant cells to blood vessel walls Iwamoto et al., 1996.. Therefore, by covalently linking multiple copies of an anti-tumor peptide the binding afnity has been Peptide dendrimers have proven to be useful substitutes for proteins and even DNA in a number of autoimmune disease model systems. Dendrimers based on peptides from various proteins have been used to generate autoimmune conditions in normal animals that are comparable to the condition systemic lupus erythematosus SLE. James and Harley, 1998; Mason et al., 1999; Farris et al., 1999.. Mason showed that inoculation of mice with a MAP consisting of eight copies of a peptide derived from the spliceosomal Sm protein resulted in epitope spreading, a response usually attributed to an immune response against self proteins and DNA. Anti-nuclear antibodies were detected in some animals illustrating that the MAP acted as a protein and DNA mimetic. Putterman and Diamond 1998. identied a peptide mimetic for double stranded DNA that elicited anti-DNA antibody production. Mice developed a lupus-like syndrome creating an experimental model for SLE. Using dendrimers Khalil et al. 2001. replicated the T cell-dependent nature of SLE using a peptide immunogen instead of a DNA immunogen and characterized the T cell response. Unlike other examples mentioned thus far, it was determined that the increased response to the dendrimer was dependent more on the amino acid sequence than on valency. A plausible explanation for the observed results is that the presence of the dendrimer backbone allowed more effective processing and presentation of the correct T helper epitopes. During a similar study where the specicity of antibodies was examined, it was found that dendrimers themselves could not only bind to antibodies but also directly interact with the 60-kDa Ro nuclear protein in various assays Scoeld et al., 1999.. Furthermore, it was concluded that Ro and La, components of a single ribonucleoprotein particle, directly interact. The protein-binding sequences within Ro were identied using den-
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drimers; the dendrimers were an important tool for exploring both intramolecular and intermolecular protein-protein interactions. Even though the afnity of interaction may not be absolutely determined because the use of dendrimers may increase avidity in immunoassays by several logs Tam, 1996., the relative afnities may be determined. In the context of SLE, the peptide dendrimer provides useful reagents for probing the extent of immune tolerance in the B and T cell compartments to nuclear antigens. The weak antimicrobial activity of monomeric peptides has been observed to increase after multimerization. An 11-residue peptide representing a sequence from human lactoferrin an iron-binding glycoprotein found in neutrophils and secretions. was incorporated into peptide dendrimers consisting of 1, 2, 4, 8 or 16 copies and the dendrimers were tested for antibacterial activity Azuma et al., 1999.. An increase in antimicrobial activity correlated with an increase in the peptide copy number. The dendrimer containing 16 copies of the peptide was as effective as the traditional antibiotic gentamycin against Gram-negative and Gram-positive bacteria. In this case, the authors suggest that the monomeric peptide is too short to span the membrane and create an ionic pore whereas the dendrimers are large enough to span the membrane and form a pore or mimic pore formation thus resulting in microbicidal activity. To advance the dendritic design a step further, Lu et al. 2001. designed an inactive tetrapeptide
with a basic and hydrophobic motif derived from -strand topology of tachyplesins and protegrins which exhibited signicant antimicrobial activity when synthesised as a dendrimer. The tetrameric dendrimer is similar in molecular weight and antimicrobial potency as protegrins or tachyplesins. More importantly, each peptide dendrimer assayed was found to have higher antimicrobial activity against fungi, Gram-negative and Grampositive bacteria than linear tandem repeating peptides.
6. Peptide dendrimers as immunogens Possibly the most widely known biological application of peptide dendrimers is the use of MAPs as immunogens. Immunogens are distinguished from antigens by their ability to induce an immune response in vivo. MAPs, typically of the form shown in Fig. 2, have enormous potential as immunogens and a signicant portion of this potential has already been realized. The advantages of using MAPs as immunogens include: Simplicity in design and synthesis; Versatility for investigating various immune responses; Reliability of generating site-specic antibodies; and Generation of site-specic antibodies in the laboratory.
Fig. 2. a. Schematic representation of a multiple antigen peptide MAP., incorporating eight peptide monomers. b. An increase in the number of Lys branching units increases the number of surface amine groups.
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Since the rst report of the synthesis and potential of MAPs by Tam 1988. MAPs have been employed routinely as immunogens for antibody production and cytotoxic immune responses.1 Before we review the application of MAPs as immunogens, several points regarding peptide sequence selection and design of MAPs will be presented. 6.1. Summary of immune responses This overview aims to briey summarize the role of peptide epitopes in the induction of immune responses and to alert the reader to the importance of epitope selection. In general, there are two types of immune responses, humoral and cell-mediated. Induction of a humoral response results in the production of antibodies and relies upon the activation of both B cells and T helper cells. Cytotoxic T cells and T helper cells, on the other hand, mediate cellular immune responses. For maximal antibody production a B cell must receive two signals. It must bind a B cell epitope to the surface immunoglobulin with high afnity. The B cell epitope is usually a peptide or protein segment approximately nine residues in length. Secondly, the B cell must interact with an activated T helper cell leading to B cell maturation and production of antibody with the same specicity as the surface immunoglobulin. The T helper cell is in turn activated by interaction with another type of cell, the antigen presenting cell APC.. The role of the APC is to engulf extracellular material, including foreign peptides and proteins, and degrade the proteins and peptides into short segments. These short segments are subsequently presented on the APC surface in association with MHC class II molecules. The T helper cell is activated if it encounters an APC presenting a peptide-MHC complex that binds with high afnity to the T cell receptor TCR.. The peptide associated with the MHC molecule is referred to as a T helper epitope. Without the presence of both the B cell epitope and T helper epitope
antibody production is generally short-lived and the highly desirable memory response is lacking. Activation of T helper cells is also required for the induction of a cell-mediated response. In this case the T helper cells interact with and provide cytokines for the stimulation of cytotoxic T cells. The main role of the cytotoxic T cells is to recognise and kill host cells which are expressing nonself peptides associated with MHC class I proteins such as peptides from intracellular bacteria and viruses. These peptides are referred to as cytotoxic T lymphocyte CTL. epitopes. The majority of host cells are capable of degrading intracellular proteins and presenting CTL epitopes on the surface when bound to MHC class I molecules. On the rst occasion that a cytotoxic T cell encounters a host cell expressing a non-self epitope it undergoes maturation aided by T helper cells. and is then capable of killing other host cells expressing the same CTL epitope. In summary, humoral responses require B and T helper epitopes and cell-mediated responses require CTL and T helper epitopes. An effective synthetic immunogen must contain peptide sequences that activate the appropriate type of immune response and the exibility of the MAP system allows incorporation of multiple epitopes in to the same construct. 6.2. Design of MAPs for induction of an antibody response B cell epitopes alone are poor immunogens and have traditionally been conjugated to large carrier proteins that provide strong T helper cell epitopes. The evolution of the MAP system now allows B cell and T helper epitopes to be assembled directly onto a small polylysine core. In contrast to conventional peptide-protein conjugates where peptide epitopes account for less than 20% of the total weight, MAPs consist of concentrated epitopes and possess an immunologically silent lysine core Posnett et al., 1988; Del Giudice et al., 1990.. In some cases there is signicant difference and a generally better quality of antisera in using MAPs vs. peptide protein conjugates McLean et al., 1991; Wang et al., 1991; Estaquier et al., 1993; Molnar et al., 1993;
A recent literature survey showed that Tams 1988 paper has been cited over 600 times.
K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229 Table 3 Utilization of peptide dendrimers for antibody and CTL production Target Bacteria Tandem repeat motifs, Leishmania SLT-1 protein, Escherichia coli P30 antigen, Toxoplasma gondii Glucosyltransferase, Streptococcus Toxin T, Bordetella pertussis Toxin B, Vibrio cholera Membrane protein 1, C. trachomatis Membrane protein, Neisseria meningitidis Cyclase, Saccharomyces cere isiae Exoproteases, Pseudomonas aeruginosa Gingipains, Porphyromonas gingi alis Porin protein, N. meningitidis Virus Surface antigen, Hepatitis B virus Reductase, Herpes simplex virus VP2 and VP7, bluetongue virus VP1 protein, foot-and-mouth disease virus gp120, HIV References
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Liew et al. 1990. Boyd et al. 1991. Darcy et al. 1992., Godard et al . 1994. Smith et al. 1993., Smith et al. 1994a. Felici et al. 1993. Halimi and Rivaille 1993. Zhong et al. 1993. Christodoulides and Heckels 1994. Shi et al. 1994. Coin et al. 1997. Genco et al. 1998. Christodoulides et al. 1999.
Nef, HIV-1 Polymerase, inuenza virus Hemagglutinin, inuenza virus NSm protein, bunyamwera virus B19, parvovirus Core proteins, vaccinia virus gp46, HTLV-1 V3 loop, HIV-1 MVF, measles virus gp51, bovine leukemia virus Fusion protein, respiratory syncytial virus VP3, Hepatitis A virus V3, feline immunodeciency virus VP1 protein, JC virus Parasite CS protein, Plasmodium berghei
Tam and Lu 1989., Manivel et al. 1993. Lankinen et al. 1989. Li and Yang 1990. Francis et al. 1991., Lugovskoi et al. 1992. Brown 1992. Wang et al. 1991., Defoort et al. 1992a,b. Nardelli et al. 1992a,b, 1994., Nardelli and Tam 1993. Levi et al. 1993., Kelker et al. 1994. De Santis et al. 1994., Huang et al. 1994. Fraisier et al. 1994., Vogel et al. 1994. Estaquier et al. 1992, 1993. Nieto et al. 1992. Toth et al. 1993., Naruse et al. 1994. Zeng et al. 2000. Nakitare and Elliott 1993. Saikawa et al. 1993., Anderson et al. 1995. Vanslyke and Hruby 1994. James et al. 1995. Cruz et al. 2000. Olszewska et al. 2000a,b. Kabeya et al. 1996. Hsu et al. 1999. Pinto et al. 1998. Rigby et al. 1996. Aoki et al. 1996.
CS protein, P. malariae CS protein, P. falciparum
Romero et al. 1988., Migliorini et al. 1993. Zavala and Chai 1990., Chai et al. 1992. Widmann et al . 1992., Valmori et al. 1994. Del Giudice et al. 1990. Munesinghe et al. 1991., Pessi et al. 1991. Valmori et al. 1992., Nardin and Nussenzweig 1993. Calvo-Calle et al. 1992, 1993. De Oliveira et al. 1994., Ahlborg 1995. Nardin et al. 1998, 2001., Moreno et al. 1999.
204 Table 3 Continued. Target CS protein, P. yoelii
References Wang et al. 1995., Marussig et al. 1997. Franke et al. 2000. Del Guercio et al. 1997. Auriault et al. 1991., Khan et al . 1994. Pancre et al. 1994. Reynolds et al. 1994. Ferru et al. 1997. Argiro et al. 2000.
CS protein , P. i ax P28 antigen, Schistosoma mansoni Triose-phosphate isomerase, S. mansoni Sm28GST and sTPI, S. mansoni Sm37-GAPDH and Sm10-DLC, S. mansoni Other Trp protein, Drosophila UL47, UL8, Vmw63 and UL42 Lutropin, human Ribosomal protein, plant Fatty acid binding protein, human Transforming growth factor , human Alkyltransferase, human Cytochrome P-4501A, rat Mast cell protease-5 , mouse ATPase, plant Calcium channel, rabbit Sperm myoglobin, whale Sperm protein, human Protein kinase p34cdc2 , eukaryote Membrane domain of band 3, human Angiotensin II type-1 receptor, rat Actin-fragment kinase, plasmodia Seed protein, plant IGF binding protein, bovine Glutamate receptor, rat Glucose transporter, rat RNA-binding protein, rodent Insulin receptor, human Myotrophin, human Iron regulatory factor, human Protein G, human Tyrosine kinase, sponge Chaperonin polypeptide, eukaryote Sperm protein , Xenopus pP344 retinal cell, chicken Myosin phosphatase, chicken Prion protein, mouse Ameloginin, mouse Histones, mouse GTPase, rat Neurexins, rat T-tubule, rabbit Calcium release channel, rabbit Anion transport protein band 3, human Guanylin, human Prolin, human
Wong et al. 1989. McLean et al. 1990., Parry et al. 1993. Sinclair et al. 1994., Marsden et al. 1994. Troalen et al. 1990. Szymkowski and Deering 1990. St John et al. 1991. Lu et al. 1991. Pegg et al. 1991. Edwards et al. 1991. McNeil et al. 1992. Suzuki et al. 1992. Malouf et al. 1992. McLean et al. 1992. Vanage et al. 1992, 1994. Kamo et al. 1992. Kang et al. 1992. Zelezna et al. 1992, 1994. Gettemans et al. 1993. Monsalve et al. 1993. Arnold et al. 1993. Molnar et al. 1993. Nagamatsu et al. 1993. Henderson et al. 1993. Itoh et al. 1993. Sil et al. 1993. Emery-Goodman et al. 1993., Gray et al. 1993. Raymond et al. 1993. Schacke et al. 1994. Lingappa et al. 1994. Bauer et al. 1994. Iio et al. 1994. Shimizu et al. 1994. ORourke et al. 1994. Simmer et al. 1994. Meziere et al. 1994. Wilson et al. 1994. Perin 1994. Stout et al. 1994. Callaway et al. 1994. Crandall and Sherman 1994. Kuhn et al. 1994., Cetin et al. 1994. Finkel et al. 1994.
K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229 Table 3 Continued. Target Type V collagen, human Phosphatase 2A, human DR molecules, human Elan, human Gangliosides, human 5 -reductase 2, human Heme oxtgenase-1, human Spliceosome protein, human Cyclin, human Phospholipase A2 , human Kainate receptor, goldsh Myelin basic protein, murine Serotonin transporter, rat Ras p21, human Poly ADP-ribose. polymerase, eukaryote Seed proteins, Vigna angularis Sm BrB protein, primate pRL1a, murine leukemia N-myc oncoprotein References Moradi-Ameli et al. 1994. Zolnierowicz et al. 1994. Demotz et al. 1994. Nara et al. 1994. Helling et al. 1994. Eicheler et al. 1994. Kutty et al. 1994., Smith et al. 1994b. James et al. 1995. Digweed et al. 1995. Sa et al. 1995. Wo and Oswald 1995. Zhou and Whitaker 1996. Zhou et al. 1996. Schott et al. 1996. Duriez et al. 1997. Kajiwara and Tomooka 1998. Arbuckle et al. 1998. Manki et al. 1998. Yang et al. 1999.
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Vogel et al., 1994.. A comparative study by McLean et al. 1991. identied what has now become a general trend in the properties of antisera elicited by different constructs, sera obtained from animals inoculated with MAPs is of a higher titer and produced more rapidly than those obtained with protein conjugates which are in turn higher than those obtained with resin-linked peptides. The MAP format allows investigators to overcome the immunodominance conferred by linker and carrier proteins and elicit antibody responses to weakly immunogenic sub-dominant peptide sequences. Examples of the use of MAPs as immunogens are listed in Table 3. There are many possible arrangements of linking B and T helper epitopes on MAPs, the most popular approach to date is to link the epitopes in a tandem formation. The main drawback with this method is that new immunodominant epitopes may be formed within the tandem peptide and the MAP is not processed and presented as the intended design. One empirical solution suggested by Lo-Man et al. 1994. is to insert a longer T epitope to preserve the diversity of T cell recognition. Alternatively convergent strategies may be employed where the dendrimer contains copies of each peptide sequence on sep-
arate arms thereby decreasing the possibility of neo-epitope formation and increasing the response to the desired epitopes. Tandem, or diepitope, MAP constructs have been extensively studied in the malaria synthetic vaccine model. Ten MAP constructs were synthesized, each containing a combination of described B and T cell epitopes, the constructs were designed to evaluate the relevance of the number of copies, stoichiometry, and orientation of T and B sequences in a diepitope model Tam et al., 1990.. In this case it was concluded that there was no advantage in using octameric instead of tetrameric MAPs, and that having the B cell epitope at the N-terminal produced a more efcient immunogen. The degree of protection against challenge of immunized mice with 2000 Plasmodium berghei sporozoites correlated with the antibody levels obtained by the immunization protocol. Tetra- and octameric MAPs have been utilized in most studies to date. The number of branches required tends to depend largely on the amino acid residues: with peptides ) 15 amino acids we have found that there is no real advantage in using more than four branches. A similar conclusion was reached by Francis et al. 1991. where guinea pigs were immunized with constructs con-
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taining one, two, four or eight copies of a footand-mouth disease virus peptide. It was determined that in this case presentation in a tetrameric structure was sufcient for an optimal antibody response. Diepitope MAPs were synthesized in the design of a synthetic vaccine model for hepatitis B using a different chemical approach Tam and Lu, 1989.. A B cell epitope and T helper epitope were used in mono- and diepitope congurations, in the latter the two peptides were connected in alternating forms on the lysine core instead of in tandem. Only when the B cell epitope was presented in the MAP form with the T helper epitope was it immunogenic in rabbits. Therefore, these results conrmed that a diepitope MAP format may overcome the poor immunogenicity of a linear peptide. Mono- and diepitope MAP congurations were further analyzed in a synthetic model for the human immunodeciency virus type 1 HIV-1. Nardelli et al., 1992a.. Monoepitope MAPs consisting of four copies of a major neutralizing epitope of HIV-1 were synthesized along with diepitope MAPs that also contained a known T helper epitope at the C-terminus of the B cell epitopes. The diepitopes were immunogenic in all species tested whereas the monoepitope MAPs elicited species-specic responses. The use of universal or promiscuous T helper epitopes which are recognized by three or more strains of mice are particularly useful for diepitope constructs and have been used to enhance the immunogenicity of B cell epitopes in a number of MAPs McLean et al., 1992; Munesinghe et al., 1991; Marguerite et al., 1992.. Finally, Levi et al. 1993. have demonstrated that boosting with B cells and T helper cell epitope MAP constructs induced a higher antibody titer for HIV-1 and improved cellular responses than boosting with recombinant gp160 protein. The use of chemoselective ligation through oxime bond formation is gaining notoriety as a feasible method for immunogen production. Selfadjuvanting tetrameric MAP constructs incorporating B cell and T helper epitopes from different sources induce strong antibody responses Nardin et al., 1998; Zeng et al., 2000..
6.3. Cell-mediated responses induced by lipidated MAPs Induction of a cell-mediated response relies upon the delivery of CTL epitopes to an intracellular endosomal pathway that results in the presentation of peptides in association with class I MHC molecules. Others have found the value of lipidated peptide in eliciting cellular responses because of the increased efciency of delivery of peptides to the intracellular compartment. Longchain lipidic amino acids have been developed by Gibbons et al. 1990. in combination with detergent-solubilized liposome as a delivery system. This approach was applied to the MAP format in successful vaccine models against HIV-1 Defoort et al., 1992a,b; Nardelli et al., 1992a, 1994. and Chlamydia trachomatis Zhong et al., 1993; Pancre et al., 1994.. A cytotoxic response was induced by a lipidated MAP containing a peptide sequence from gp120 envelope protein of HIV-1, III-B isolate which overlaps a B cell epitope, a T helper epitope and a CTL epitope. This response was detected after a single inoculation without extraneous adjuvant, was superior to the response induced by a full cycle of inoculations with the non-lipidated MAP in complete Freunds adjuvant and was still detectable 7 months later. The lipidated MAP was found to produce systemic antibody and cellular responses regardless of the route of inoculation employed Nardelli et al., 1992b, 1994.. The ability of MAPs containing the lipid moiety tripalmitoyl-S-glyceryl cysteine P3 C. to induce mucosal antibody response via oral administration adds a new dimension to applications of the MAP constructs, and it may be particularly useful in preventing transmission of pathogens, such as HIV, through mucosal surfaces. 6.4. Possible reasons for increased immunogenicity The multimeric MAP approach has been shown to increase the immunogenicity of weakly immunogenic monomeric peptides, there are several possible reasons for the observed enhanced activity. The conformation of the peptide epitopes within the MAP may contribute to the longevity
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and strength of the responses induced by MAPs. MAPs attain a somewhat unnatural polymeric structure when compared to a linear peptide which may aid in resistance to in vivo proteolytic degradation leading to a longer immunogen halflife. This persistence of immunogen in the systemic circulation possibly induces longer lasting immune responses. The increase in immunogenicity could also be due to new T helper epitopes being generated within the MAP format. Even though the lysine core itself is immunologically silent the contribution of one or two residues to the attached peptide sequences could result in the production of new epitopes. This would provide a plausible explanation for the fact that the antibody response to the repetitive NANP peptide from Plasmodium falciparum CS protein is restricted to a single mouse strain when administered as a linear peptide but administration of the peptide in a MAP format results in antibody production in a further ve strains that differ in MHC haplotype Pessi et al., 1991.. Another alternative is that the MAP construct allows the desired epitopes to be processed and presented by APCs in a more efcient manner. For instance all residues in a linear peptide may be accessible to degrading enzymes, whereas the MAP conguration may actually protect certain residues from enzymatic cleavage resulting in the presentation of different epitopes. In many cases the more immunogenic diepitope MAP constructs are synthesized with the desired T helper epitope in close proximity to the branching lysine core. The design of a MAP provides a scaffold for close packing of peptide sequences that may allow the stabilization of the secondary structure and the reverse turns of peptides. Furthermore, the distal end of the peptide away from the core of MAPs is more exposed and exible than the proximal end, for these reasons we may expect the immunogenicity of the B cell epitope to be greater if the epitope is located at the distal, N-terminal site. The multimeric nature of the MAP system may also contribute to the well-documented increases in immunogenicity. The higher copy number of closely packed epitopes could lead to greater B
cell surface cross-linking through surface immunoglobulins resulting in increased activation and antibody production. 6.5. Potential application: peptide dendrimers as synthetic accines The majority of vaccines currently approved for human use consist of either heat-killed or liveattenuated infectious agents. Despite the successes of such vaccines, development by conventional methods is limited by several factors, namely: Hazardous production; Cold chain required for storage; Presence of contaminating materials; Risk of reversion to infectious state; and Side effects of vaccination. Subunit vaccines consisting of either whole recombinant proteins or synthetic peptides are appealing alternatives because they are safe, selective and chemically dened. Peptides have further advantages in that short peptide sequences may be synthesized that only contain the epitope of interest and deleterious epitopes may be omitted. Large quantities of chemically puried peptide vaccines can be prepared with automated methods and peptide-based immunogens are more likely to be resistant to denaturation, and they can be easily stored and transported without refrigeration. A peptide dendrimer malaria vaccine recently entered a Phase I study in human volunteers Nardin et al., 2001.. The vaccine was assembled using chemoselective ligation via oxime bonds and consists of four peptide branches each containing B cell epitopes and a promiscuous T helper cell epitope from P. falciparum CS protein. Exogenous adjuvant was not required due to the inclusion of a P3 C moiety covalently linked to the polylysine core. The majority of the inoculated volunteers developed high titered antibody responses as well as T cells specic for the T helper epitope. The use of MAPs as vaccines may extend past the use of relatively small tetra- and octameric
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constructs that contain a minimal quantity of epitopes. MAP technology lends itself to the design of macromolecular structures that may mimic whole organisms or several different organisms. Ultimately several similar or different closely packed peptide immunogens prepared as MAPs could be anchored on lipid vesicles that mimic surface proteins. Immunogenicity could be broadened and enhanced by the inclusion of a non-covalent mixture of B cell and T helper and CTL epitopes from various proteins. At the same time the combination of adjuvant effects of liposome and the built-in lipid anchor may replace the need for additional adjuvant. Lipidated MAPs have been shown to become entrapped in liposomes in an efcient manner, nearly 80% of
lipidated MAPs were incorporated as compared to 2 5% without the lipid anchor Defoort et al., 1992a,b. in one example.
7. Synthesis overview Synthesis of peptide dendrimers embraces a broad range of chemistry from conventional solid-phase peptide synthesis schemes in organic solvents to the formation of regiospecic amide or non-amide bonds in aqueous solutions. Strategies for preparing peptide dendrimers can generally be divided into two categories, namely the divergent and convergent approaches Fig. 3.. The divergent strategy is a direct approach by which
Fig. 3. A schematic comparison of divergent and convergent synthesis methods.
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the dendrimer is built stepwise and diverges outward. Stepwise solid-phase synthesis has been used to prepare the dendrimer in a continuous operation on a solid support. However, the convergent strategy is an indirect, modular approach by which peptidyl surface functional groups and the branching unit are prepared separately. The
puried components are then linked together and peptide sequences converge to the branching unit as a dendrimer. Speed and efciency are advantages of the divergent stepwise strategy particularly by solidphase methods because intermediates are neither puried nor characterized. This strategy is suit-
Fig. 4. Examples of simple amino, carboxylic and hydroxylic cores.
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able for validation of concepts and for small peptide dendrimers that may be separated from their byproducts by powerful systems. In many cases stepwise synthesis is not practical, either because of the nature of the branching unit preventing integration into a stepwise strategy or because the resulting dendrimer is too large e.g. ) 15 kDa. to be separated from its byproducts with condence. An advantage of the convergent strategy is chemical unambiguity because the protected or unprotected peptide segments and the branching unit used in coupling reactions are puried prior to reaction, thereby limiting the range of byproducts and facilitating purication. A disadvantage lies in the operational complexity: the convergent strategy requires more steps than stepwise synthesis, including additional purication and characterization steps for intermediates. The convergent strategy could also pose signicant problems with regard to the solubility of peptide intermediates when protected peptide segments are being used. Thus, a recent trend has developed in which aqueous, soluble unprotected peptides are used that are convenient to handle, purify and characterize. Several laboratories have made extensive efforts to develop new methodologies of ligation chemistry that involve the fewest steps possible. These chemistries are based on chemoselectivity and orthogonality of unprotected intermediates for assembling peptide dendrimers. 7.1. Purication and characterization Methods for purication and characterization of peptide dendrimers are similar to those of peptides and proteins. Crude synthetic peptide dendrimers derived from the divergent strategy often require multiple methods that may include sequential steps of dialysis or gel-ltration chromatography, followed by RP-HPLC or high-performance ion-exchange chromatography. Even by these steps it may still not be possible to remove byproducts with a single modication or deletion of an amino acid from the desired product. Synthetic products assembled using a convergent strategy can be rened by most chromatographic
methods resulting in a homomeric product. Characterization methods for peptide dendrimers include the usual panel of techniques: amino acid analysis; SDS-PAGE; capillary zone electrophoresis; and enzymatic digestion. Mass spectrometric analysis has now become an indispensable tool for determining the molecular weights indicating product homogeneity of these complex molecules. 7.2. Types of dendrimer cores and branching units For the design of articial proteins the type of core selected is critical because it may confer rigidity and exert conformational inuence on the overall structure of the peptide dendrimer. Elaborate cores Unson et al., 1984; Sasaki and Kaiser, 1989; Ghadiri and Case, 1993; Chapman et al., 1994; Schneider and Kelly, 1995; Tsikaris et al., 1996; Wrighton et al., 1997; Fairlie et al., 1998; Frey, 1998; Goodman et al., 1998; Jefferson et al., 1998. are designed to induce or to assist folding into structural motifs such as -helices, -sheets, reverse-turns and loops which are often lacking in peptide monomers. However, most cores employed to date are simple small organic molecules - 1000 Da with the sole purpose of clustering or amplifying peptides to enhance the polyvalency effect. Selected examples are shown in Fig. 4. Such cores are often commercially available allowing many groups to investigate the use of peptide dendrimers in various applications. The most common cores are simple amino compounds, namely amino acids and dipeptides. The simplest amino cores are the trisethylene amine. ammonia types rst used in organic dendrimers that have recently been utilized in formation of peptide dendrimers. The use of heterocyclic compounds such as porphyrin and constrained and unusual amino acids has also been reported Sasaki and Kaiser, 1989.. Simple organic polycarboxylic acids have been used as branching units Roth and Heidemann, 1980; Kemp and Petrakis, 1981; Thakur et al., 1986; Fairlie et al., 1998; Goodman et al., 1998., however, many branching units contain a combination of functional groups. As a group, carbohydrates and their intermediates represent a diverse and attractive source of polyhydroxylic branching
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units because of their small sizes and the stereochemical variations of pendant hydroxyls Newkome et al., 1991; Whittaker et al., 1993; Murer and Seebach, 1995.. In addition, their hydrophilic nature favors convergent synthesis in aqueous solutions. The utility of hydroxylic and carboxylic branching units is currently limited by the need for a synthetic methodology to distinguish between these groups that also occur frequently in peptides. In the case of hydroxylic groups, they are further hindered by the less reactive nature when compared to amines and carboxylic acids. As a result, their use has not yet been widely explored in peptide dendrimers. 7.3. Polylysine branching units Polyamino acids consisting of several branches of a trifunctional acid represent the largest group of branching units being used today Tam, 1988; Rao and Tam, 1994.. By virtue of their trivalency, reactivity and synthetic expedience branching units derived from lysine and its homologs are the most popular. Lysine with N - and N -amino groups as reactive ends is a particularly suitable trifunctional amino acid to form a branching unit by a limited sequential propagation of lysines. These low molecular weight cores with 2 n reactive amino ends can then serve as attachment sites for peptides. Solid phase schemes are the method of choice for the synthesis of lysine
branching units Tam, 1988; Tam and Lu, 1989. where di-protected Lys, e.g. Boc LysBoc. or Fmoc LysFmoc., is used to produce a core of multiple levels of lysines. The diamino nature of lysine creates a situation where each additional level of Lys effectively doubles the number of sites to which peptide monomers may be attached. A typical branching lysine unit consists of three lysines to give a tetravalent peptide dendrimer Fig. 5.. Further divergence of the K 2 K unit to one or two additional levels will generate di-K 2 K and tetra-K 2 K dendrons with reactive ends of 8 and 16 amino groups, respectively, to which peptides can be attached. Peptides may be synthesised directly onto the lysine dendron using a divergent strategy or dendrons can be further functionalized with electrophiles and thiol nucleophiles for convergent ligation of peptides. The K 2 K-type branching unit is small in size when compared with the bulk of peptides layered around the surface. As an example, a dendrimer consisting of a branched unit and eight copies of peptide chains, each containing 14 amino acid residues, would have an average molecular weight of 13 14 kDa. Only a small fraction of the mass is due to the lysine unit; 93% of the mass is due to peptide sequences. Many variations of K 2 K cores have evolved. For example, a version containing a -alanine spacer Huang et al., 1994. at the -amine of Lys has produced a branching unit that has symmetri-
Fig. 5. Polylysine cores consisting of K 2 K and di K 2 K, where Xaa can be either OH, an amino acid, peptide or organic core see Fig. 4..
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Fig. 6. Different K 2 K, polyamino, linear and cyclic cores used for dendrimer assembly. Ahx s aminohexanoic acid.
cal branches. In another example, in order to limit the exibility of the branched lysyl template, ornithine, the lower homolog of Lys, as well as more constrained diamino acids have been used Roth and Heidemann, 1980; Hahn et al., 1990.. Non-branched linear polylysine cores Tsikaris et al., 1996. and cyclic peptides Goodman et al., 1998. have also been developed to impart different forms of dendrimeric architectures Fig. 6.. As eluded to earlier, the basic branching unit may be further modied to allow peptide attachment. As shown in Fig. 7, attachment of serine to the K 2 K dendron during solid-phase synthesis allows the production of a branching unit with N-terminal aldehyde moieties. This serine can be
readily converted to aldehyde at neutral pH in aqueous conditions and has been used by Spetzler and Tam 1995. as a convenient masked aldehyde precursor. Thiol-functionalized branching units are popular and convenient to prepare because of the relative ease with which reactive thiol moieties may be placed on peptide units in preparation for convergent syntheses Lu et al., 1991; Defoort et al., 1992b; Zhang and Tam, 1997b.. Drijfhout and Bloemhoff 1991. introduced a thiol-functionalized branching unit based on the K 2 K dendron by coupling N- S-acetylmercaptoacetyl.-glutamyl residues to the amino groups. Subsequently, Baleux and Dubois 1992. prepared a thiolate branching unit based on a similar design by coupling Cys to the amino groups Fig. 8.. 7.4. Di ergent stepwise strategy for peptide dendrimer synthesis Divergent methods used to prepare peptide dendrimers are operationally similar to the stepwise solid-phase syntheses of linear peptides rst developed by Merrield 1963.. Thus, protecting
Fig. 7. Solid phase synthesis of a K 2 K glyoxylyl core.
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Fig. 8. Synthesis of thiol-functionalized S-acetyl and CysNpys. di-K 2 K cores.
group schemes, resin supports, and cleavage methods found in peptide synthesis protocols are generally applicable to the divergent synthetic schemes of peptide dendrimers. Because this approach produces fully characterized dendrimers in a short time, often within a week, it is popular in biological research in which peptide dendrimers are used as tools and reagents for validating biochemical goals. Syntheses generally commence with the branching unit which is usually of the K 2 K type. In such cases two or three levels of diverging lysine are formed rst by coupling a suitable di-protected lysine, either a di-Boc or di-Fmoc derivative, to a small amino acid attached to a resin support to form a branched structure. The peptide sequence is then synthesized stepwise on this branched-lysine scaffolding. After peptide synthesis the peptide dendrimer is cleaved from the resin support, puried and characterized. Goodman Goodman et al., 1998; Jefferson et al., 1998. introduced a different divergent synthesis approach. In the synthesis of collagen models consisting three copies of homomeric peptide chains, solid-phase schemes began with monomers and then the Kemp tricarboxylic acid KTA. template Kemp and Petrakis, 1981. was used to link three monomers to form the peptide dendrimers Fig. 9.. Interchain aggregation may occur within the branched structure of dendrimers because the quantity of peptide monomers obviously increases with each additional level of lysine units. To
minimize this problem the divergent synthesis often starts with a low amino acid loading of approximately 0.1 mmolrg on a resin support. This is signicantly lower than the customary loading, typically 0.3 0.8 mmolrg resin, conventionally used in solid-phase peptide synthesis. Formation of interchain hydrogen bonds that occlude coupling reactions can also be minimized by using specic solvent combinations as well as coupling at elevated temperatures Tam and Lu, 1995.. Following synthesis and cleavage some peptide dendrimers have exhibited an unusual tendency to aggregate after cleavage from the resin due to the entrapment of aromatic scavengers. Dialysis under basic, denaturing conditions using 8 M urea is useful to remove such unwanted additives. After dialysis either RP-HPLC or high performance gel-ltration chromatography can be used to purify peptide dendrimers. A variation of the divergent synthesis scheme is the synthesis of two dissimilar peptide chains on different arms of a lysine branching unit. This is a particularly useful method for the production of immunogens containing different peptide epitopes. Methods to distinguish between - and -amines of lysine for such a purpose are based
Fig. 9. Synthesis of collagen models with three copies of homomeric peptide chains on the Kemp tricarboxylic acid core.
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on orthogonal protecting group schemes. Suitable protecting group combinations include: i. Boc Fmoc Merrield, 1963.; ii. Fmoc Aloc Kates et al., 1993.; iii. Fmoc Dde Bycroft et al., 1993.; and iv. Npys Fmoc Hahn et al., 1990; Ahlborg, 1995; Rajagopalan et al., 1995.. In general, the combination of Boc Fmoc chemistry is convenient because the protected amino acids are readily available. This chemistry utilizes both acidand base-driven deprotection methods Tam and Lu, 1989. in repetitive steps that elongate the two dissimilar peptide chains. In the K 2 K core, for example, when half of the reactive lysine amino chains e.g. -amino groups. are chemically blocked during synthesis it is possible to construct a peptide dendrimer containing two different peptide sequences Fig. 10.. The rst peptide chain can be assembled on the N -termini of the lysine matrix using Boc chemistry while the Fmoc groups of N -termini are not affected during the Boc synthesis. When the rst peptide is complete the second peptide chain can be synthesized on the N -termini using Fmoc chemistry. The heteromeric peptide dendrimer is then cleaved from the resin by HF that will also remove both the benzyl- Boc chemistry. and tertbutyl- Fmoc chemistry. protecting groups. In recent years the use of the Boc protecting group has become less popular because special apparatus must be used for deprotection by HF. Alternative methods have been developed and these include Boc substitution with i. Aloc that is removable by Pd o in the presence of a nucleophile such as morpholine, ii. Dde which is removable by 2% hydrazine in DMF, and iii. Npys 3-nitropyridino-2-sulfhydryl. which is removable under neutral conditions by a trivalency phosphine or a thiol. An example of the utilization of orthogonal protection was reported by Hahn et al. 1990. with the divergent synthesis of a chymotrypsin-like articial enzyme using three types of protecting groups on a Lys 2 Orn branching unit to form a peptide dendrimer consisting of four dissimilar chains. The rst two chains were assembled by alternating Boc and Npys chemistries on the differentially protected dipeptide Npys LysBoc. OrnFmoc.. By terminating each chain with an
Fig. 10. Schematic illustration of the synthesis of heteromeric peptide dendrimers on a di-K 2 K core.
acetylated amino acid and repeating the same chemistry, the next two chains were assembled on Boc LysNpys. coupled to the Orn side chain after the removal of Fmoc group. The strategic simplicity and exibility of divergent orthogonal synthesis also permits the incorporation of unusual moieties such as lipid chains and lipidic P3 C into the dendrimeric design Defoort et al., 1992b..
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7.5. Con ergent synthesis strategies for peptide dendrimers Convergent strategies differ from the divergent methods in that the branching unit and peptides are synthesized separately, puried individually and then coupled to form the dendrimer. Two different convergent strategies are typically used; one strategy exploits a protection scheme while the other does not. The choice of a convergent strategy is often arbitrary. Consideration should, however, be given to the nature of the peptide monomer, the complexity of the peptide dendrimer, and the expertise of a given laboratory. An additional factor is the orientation of the peptide that is to be attached and this is governed by the functional consequences. For example, peptides derived at or near C-termini can be arranged as in the native protein by linkage through the N-terminus to the branching unit. A variety of functional groups have been used in the development of numerous ligation protocols in order to provide site-specic and chemically unambiguous dendrimers. A detailed review of different ligation strategies and methodologies was published recently by Tam 2000.. 7.6. Con ergent synthesis with a protecting group scheme The convergent synthesis using protecting groups Blake, 1981; Yajima and Fujii, 1981; Yamashiro and Li, 1988; Hojo and Aimoto, 1992; Sakakibara, 1995; Albericio et al., 1997. generally follows conventional segment coupling schemes with enthalphic activation by a coupling reagent. The major difference between forming a branched dendrimer and forming a linear peptide by segment condensation is that two or more protected peptide segments are simultaneously coupled to a core to afford the former. Purication by gel permeation chromatography and RP-HPLC is facilitated by the substantial differences in the molecular weights of the desired product compared to incompletely coupled byproducts. Sasaki and Kaiser 1989. reported formation of such a peptide dendrimer using convergent synthesis where an articial hemeprotein containing four
Fig. 11. Convergent synthesis of an articial hemeprotein.
identical peptides tethered to a coproporphyrin core was assembled. The 15aa fully protected peptide segment was synthesized through a twosegment condensation 8aa q 7aa. and then coupled to the hydroxysuccinimide ester of the branching unit through its N-terminal Fig. 11.. A common approach is to use a less elaborate chemistry as illustrated by McLean et al. 1992. who described a rapid convergent scheme for the formation of MAPs. To obtain a 17aa protected peptide segment they used an Fmoc-tertbutyl protecting group scheme with a highly acid-labile resin. The cleaved protected peptide containing a free carboxylic acid was then coupled to a tetrameric K 2 K branching unit. However, the poor solubility of this protected peptide resulted in varying degrees of coupling in six MAPs, ranging from complete to no observed coupling. Although this difculty does not impede the biological goals it certainly indicates the unpredictable nature of using protected peptide segments in convergent schemes. 8. Chemoselective ligation strategies: unprotected peptide synthesis A recent advance in convergent synthesis has been the use of unprotected peptide segments
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instead of bulky protected peptides. One such strategy that links unprotected peptide monomers to a branching unit to form a dendrimer via non-amide bonds is regiospecic chemoselective ligation. This type of ligation exploits a pair of mutually reactive functional groups, usually a nucleophile and an electrophile, on the peptide and branching unit that only react specically with each other. Another strategy that links unprotected peptides via amide bond is orthogonal ligation Tam et al., 2000.. Orthogonal ligation is N-terminal specic, thus, a peptide containing an N-terminal Lys can be ligated in the presence of other N-terminal amino acid peptides. An advantage of these types of ligation is their specicity because peptide segments and branching units are used in their unprotected state they are more likely to be water soluble and amenable to purication by RP-HPLC. A plethora of chemoselective and orthogonal ligation methods and many variations have been developed Tam, 2000.. Furthermore, the nucleophile and electrophile can be placed on either the peptide segment or the branching unit creating numerous potential combinations. In addition, there are many readily available bifunctional groups that are used to modify K 2 K dendrons in preparation for ligation including: succinimide esters; maleimides; sulfenyl thiols; and chloroacetyl groups Keller and Rudinger, 1975; Carlsson et al., 1978; King et al., 1978, 1986; Yoshitake et al., 1979; Wunsch et al., 1985; Ottl et al., 1999.. Examples of the types of bonds that may be formed by regiospecic chemoselective ligation include hydrazone, oxime, thioether and thioester bonds. Each chemoselective, or orthogonal, strategy consists of an initial capture step to form a covalent intermediate between two peptide segments. Then, an amide bond is formed via an intramolecular acyl transfer resulting in the coupling of the two different segments. The strategies are categorized by two major chemistries: thioester and imine. 8.1. Thioester ligation In the thioester ligation strategy a thioester and an N-terminal cysteine are required. The
initial capture product is a thioester and an S, Nacyl shift results in formation of a cysteine residue at the ligation site Muir et al., 1994; Tam et al., 1995.. Other variations of cysteine ligation such as perthioester ligation have also been developed to make thioester a popular orthogonal ligation strategy Tam et al., 2001.. 8.2. Imine and imide ligation The imine chemoselective ligation strategy requires the presence of an acyl-aldehyde and an N-terminal weak amine. Commonly used imine ligation strategies include oximation, hydrazone formation and imide ligation. Oximation has been used in protein conjugation reactions Vilaseca et al., 1993; Rose, 1994. and in the synthesis of peptide dendrimers Shao and Tam, 1995; Pallin and Tam, 1996.. Hydrazone formation has also been employed in ligation reactions through the reaction of aldehyde and C-terminal hydrazide Gaertner et al., 1992.. For orthogonal imide Nterminal. ligation a C-terminal glycoaldehyde ester is required for reaction with either an Nterminal NT. cysteine pseudoproline. or NTserine or NT-threonine oxaproline.. The mechanism of imide ligation is similar to orthogonal thioester ligation with an imine capture, a ring closure to a thiazolidine NT-Cys. or oxazolidine NT-SerrThr. and then an S, N- or O, N-acyl shift to form an imide Liu and Tam, 1994.. Pseudoproline ligation has been successfully employed in the production of synthetic HIV-1 protease analogs Liu et al., 1996.. Each of these imine or imide ligations relies on the presence of aldehydes and since aldehydes are not found in amino acids several methods have been used to introduce aldehydes to amino acids. Two convenient methods are: 1. conversion of N-terminal Ser and Thr residues to aldehyde through sodium metaperiodate oxidation; and 2. conversion of masked acetals attached to amino groups. to aldehyde by TFA or concentrated HCl. The conversion of N-terminal Ser and Thr residues to aldehydes has been used extensively in protein conjugation and designing new proteins as well as for immunological applications Zhang and Tam, 1996; Zhang et al., 1998..
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Even though the two reactive species may be placed on either the branching unit or monomeric peptide, for synthetic convenience weak bases are often added to the N-terminal of a peptide and in such cases the peptide sequence is oriented with the N-terminal residues proximal to the branching unit. The reverse orientation may be achieved with a C-terminal hydrazide derivative Rao and Tam, 1994; Shao and Tam, 1995; Pallin and Tam, 1996; Zhang and Tam, 1996; Liu and Tam, 1997.. Combinations of two, and potentially more, reaction conditions can be exploited for ligating dissimilar peptide chains to a branching unit to form heteromeric peptide dendrimers Liu et al., 1999. in a process termed tandem ligation. 8.3. Tandem cyclization strategies for peptide dendrimers Constrained peptide dendrimers have been developed in an effort to generate molecules that more closely mimic native protein conformations and to improve biological activity. One such design contains multiple closed-chain branches which are cyclic peptides constrained by end-toend, end-to-side, or side-to-side linkages Liu and Tam, 1997; Spetzler and Tam, 1996; Zhang and Tam, 1997a,b.. A general approach to achieve such constrained peptide dendrimers is to attach multiple copies of a cyclic peptide to a branching unit. Conventionally, this scheme of convergent synthesis requires two types of manipulations: a cyclization step on a linear precursor and a coupling step for its attachment to a branching unit. This two-step process usually requires a multitiered protecting group scheme. To minimize the protection and deprotection procedures of con-
ventional convergent methods and also difculties due to the insolubility of intermediates and products, simplied tandem chemoselective methods have been developed based on unprotected peptide precursors in aqueous conditions. 8.4. Di ergent synthesis and chemoselecti e cyclization Spetzler and Tam 1996. developed a method for preparing peptide dendrimers with end-to-side chain cyclic peptide monomers that combines on-resin synthesis and off-resin cyclization. First, a divergent strategy of stepwise solid-phase synthesis was used to assemble the peptides linked to a K 2 K branching unit. This branched precursor was then cleaved from the resin and its protecting groups removed to permit an off-resin cyclization that formed the constrained dendrimer through a thiazolidine cyclization in an aqueous solution Fig. 12.. The use of this method may, however, be restricted by the competing intramolecular cyclizations among the peptide branches. Ring-chain tautomerization may be used to overcome this limitation where mutually reactive nucleophile electrophile pairs simultaneously self-assemble to form peptide dendrimers with multiple copies of cyclic peptides. 8.5. Chemoselecti e cyclization and con ergent ligation Using a slightly different approach, Pallin and Tam 1995. synthesized a constrained peptide dendrimer by rst cyclizing the monomeric peptide and then attaching the monomers to a branching unit as shown in Fig. 13. The linear
Fig. 12. Use of divergent synthesis and thiazolidine intramolecular cyclization to generate a peptide dendrimer.
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sequence KAKGIGPGRAFGK- Ala derived from the neutralizing determinant of the V3 loop of gp120 was selected and prepared by Fmoc chemistry resulting in the precursor KCys. AKO NH 2 .GIGPGRAFGKSer.- Ala. The Ser moiety attached to the rst Lys served as a masked aldehyde and was oxidized to an aldehyde. A cyclic peptide was formed by utilizing the intramolecular oxime formation from the reaction between the Lys-side chain tethered O-alkylhydroxylamine and the aldehyde. The thiol residue of Cys on the side chain of the third Lys moiety was then liberated and used to ligate to
the aldehyde moieties of a glyoxyyl di-K 2 K branching unit form a peptide dendrimer containing four copies of side-to-side chain cyclic peptides. This method for formation of side-to-side chain cyclic peptides differs from the previous example Fig. 12. in the form of cyclic peptide constraint and method of synthesis. 8.6. Orthogonal cyclization and con ergent thiol ligation Tandem ligation is a strategy consisting of two or more chemoselective and orthogonal ligations
Fig. 13. Synthesis of a cyclic peptide dendrimer through side-chain to side-chain oxime cyclization and thiazolidine ligation.
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that do not require a deprotection step in the synthetic scheme. Thus, tandem ligation represents a convergent scheme with the fewest synthetic steps and the most rapid approach for synthesis of high molecular weight peptides. Three tandem ligation methods for preparing constrained peptide dendrimers have been developed based on orthogonal cyclization and convergent thiol ligation Liu and Tam, 1997; Zhang and Tam, 1997b.. These are: cysteine-thioester cyclization and thioalkylation; perthioester cyclization and thiol addition; and imine cyclization and thioalkylation. A complete discussion of these methods is found in Tam 2000.. Even though orthogonal cyclization and convergent ligation methods also use a linear unprotected peptide precursor and a functionalized branching unit as building blocks for peptide dendrimers there are two major differences between these and the previously mentioned chemoselective ligation strategies. Firstly, orthogonal cyclization methods produce lactams and secondly, only one weak base nucleophile, fullls the function of two nucleophiles in chemoselective ligation. The rst reaction common to the three reaction schemes is the orthogonal lactamization of the N-terminal nucleophile and the C-terminal electrophile of the linear precursor to generate an end-to-end cyclic peptide Zhang and Tam, 1997b; Tam and Lu, 1998; Tam and Yu, 1998; Tam et al., 1999.. The second reaction ligates the cyclic peptide monomer to a functionalized branching unit through its thiol side chain resulting in a peptide dendrimer Liu and Tam, 1997; Zhang and Tam, 1997b.. These two reactions can be performed in tandem without purication when homogeneous intermediates are used. An advantage of these methods is their high efciency because no protection or deprotection steps are involved. Racemization is minimized because there is no enthalpic activation. Furthermore, the cyclization can occur at high peptide concentrations due to the high effective molarity of an intra- vs. inter-molecular reaction. The variation in the strategies now available for the preparation of peptide dendrimers allows investigators to select a method most suitable for their individual needs. The high specicity and
homogeneity conferred by convergent chemoselective methods may be benecial in some situations whereas the rapid production of divergent peptide dendrimers could be advantageous in others. The use of peptide dendrimers as biochemical reagents and immunogens is entering a new phase. The reliability and versatility of the dendrimer system are now well-documented and investigators are looking towards the utilization of peptide dendrimers as gold-standard reagents and as a new generation of totally synthetic vaccines. References
Adesida, A.B., Aojula, R.R., Aojula, H.S., Clarke, D.J., 1999. Nonepitopic antibody binding sequence: implications in screening and development of peptide vaccines. Vaccine 18, 315 320. Ahlborg, N., 1995. Synthesis of a diepitope multiple antigen peptide containing sequences from two malaria antigens using Fmoc chemistry. J. Immunol. Methods 179, 269 275. Albericio, F., Lloyd-Williams, P., Giralt, E., 1997. Convergent solid-phase peptide synthesis. In: Fields, G.B. Ed.., Methods in Enzymology, 289. Academic press, New York, pp. 313 336. Anderson, S., Momoeda, M., Kawase, M., Kajigaya, S., Young, N.S., 1995. Peptides derived from the unique region of B19 parvovirus minor capsid protein elicit neutralizing antibodies in rabbits. Virology 206, 626 632. Andre, S., Cejas Ortega, P.J., Perez, M.A., Roy, R., Gabius, H.-J., 1999. Lactose-containing starburst dendrimers: inuence of dendrimer generation and binding-site orientation of receptors plantranimal lectins and immunoglobulins. on binding properties. Glycobiology 9, 1253 1261. Aoki, N., Mori, M., Kato, K. et al., 1996. Antibody against synthetic multiple antigen peptides MAP. of JC virus capsid protein VP1. without cross reaction to BK virus: a diagnostic tool for progressive multifocal leukoencephalopathy. Neurosci. Lett. 205, 111 114. Arbuckle, M.R., Gross, T., Scoeld, R.H. et al., 1998. Lupus humoral autoimmunity induced in a primate model by short peptide immunization. J. Investig. Med. 46, 58 65. Argiro, L., Henri, S., Dessein, H., Kouriba, B., Dessein, A.J., Bourgois, A., 2000. Induction of a protection against S. mansoni with a MAP containing epitopes of Sm37-GAPDH and Sm10-DLC. Effect of coadsorbtion with GM-CSF on alum. Vaccine 18, 2033 2038. Arnold, D.R., Moshayedi, P., Schoen, T.J., Jones, B.E., Chader, G.J., Waldbillig, R.J., 1993. Distribution of IGF-I and -II, IGF binding proteins IGFBPs. and IGFBP mRNA in ocular uids and tissues: potential sites of synthesis of IGFBPs in aqueous and vitreous. Exp. Eye Res. 56, 555 565.
220
K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229 against ribosomal P proteins: use of a multiple antigen peptide as the coating agent in ELISA. J. Immunol. Methods 179, 193 202. Carlier, E., Mabrouk, K., Moulard, M. et al., 2000. Ion channel activation by SPC 3 , a peptide derived from the HIV-1 gp120 V3 loop. J. Peptide Res. 56, 427 437. Carlsson, J., Drevin, H., Axen, R., 1978. Protein thiolation and reversible protein-protein conjugation. N-Succinimidyl 32-pyridyldithio.propionate, a new heterobifunctional reagent. Biochem. J. 173, 723 737. Cetin, Y., Kuhn, M., Kulaksiz, H. et al., 1994. Enterochromafn cells of the digestive system: cellular source of guanylin, a guanylate cyclase-activating peptide. Proc. Natl. Acad. Sci. USA 91, 2935 2939. Chai, S.K., Clavijo, P., Tam, J.P., Zavala, F., 1992. Immunogenic properties of multiple antigen peptide systems containing dened T and B epitopes. J. Immunol. 149, 2385 2390. Chapman, T.M., Hillyer, G.L., Mahan, E.J., Shaffer, K.A., 1994. Hydraamphiphiles novel linear dendritic blockcopolymer surfactants. J. Am. Chem. Soc. 116, 11195 11196. Choi, J.S., Lee, E.J., Choi, Y.H., Jeong, Y.J., Park, J.S., 1999. Polyethylene glycol.-block-polyL-lysine. dendrimer: novel linear polymerrdendrimer block copolymer forming a spherical water-soluble polyionic complex with DNA. Bioconjug. Chem. 10, 62 65. Christodoulides, M., Heckels, J.E., 1994. Immunization with a multiple antigen peptide containing dened B- and T-cell epitopes: production of bactericidal antibodies against group B Neisseria meningitidis. Microbiology 140, 2951 2960. Christodoulides, M., Rattue, E., Heckels, J.E., 1999. Effect of adjuvant composition on immune response to a multiple antigen peptide MAP. containing a protective epitope from Neisseria meningitidis class 1 porin. Vaccine 18, 131 139. Coin, D., Louis, D., Bernillon, J., Guinand, M., Wallach, J., 1997. LasA, alkaline protease and elastase in clinical strains of Pseudomonas aeruginosa: quantication by immunochemical methods. FEMS Immunol. Med. Microbiol. 18, 175 184. Crandall, I., Sherman, I.W., 1994. Antibodies to synthetic peptides based on band 3 motifs react specically with Plasmodium falciparum human malaria.-infected erythrocytes and block cytoadherence. Parasitology 108, 389 396. Cruz, L.J., Quintana, D., Iglesias, E. et al., 2000. Immunogenicity comparison of a multi-antigenic peptide bearing V3 sequences of the human immunodeciency virus type 1 with TAB9 protein in mice. J. Pept. Sci. 6, 217 224. Darcy, F., Maes, P., Gras-Masse, H. et al., 1992. Protection of mice and nude rats against toxoplasmosis by a multiple antigenic peptide construction derived from Toxoplasma gondii P30 antigen. J. Immunol. 149, 3636 3641. Dat, M.H., Behr, C., Jouin, H., Baleux, F., Mercereau-Puijalon, O., Dubois, P., 2000. Mimicking a conformational B
Auriault, C., Wolowczuk, I., Gras-Masse, H. et al., 1991. Epitopic characterization and vaccinal potential of peptides derived from a major antigen of Schistosoma mansoni Sm28 GST.. Pept. Res. 4, 6 11. Avila, S.L., Goldberg, A.C., Arruk, V.G. et al., 2001. Immune responses to multiple antigen peptides containing T and B epitopes from Plasmodium falciparum circumsporozoite protein of Brazilian individuals naturally exposed to malaria. Parasite Immunol. 23, 103 108. Azuma, M., Kojima, T., Yokoyama, I. et al., 1999. Antibacterial activity of multiple antigen peptides homologous to a loop region in human lactoferrin. J. Pept. Res. 54, 237 241. Baleux, F., Dubois, P., 1992. Novel version of multiple antigenic peptide allowing incorporation on a cysteine functionalized lysine tree. Int. J. Peptide Protein Res. 40, 7 12. Bauer, D.W., Murphy, C., Wu, Z., Wu, C.H., Gall, J.G., 1994. In vitro assembly of coiled bodies in Xenopus egg extract. Mol. Biol. Cell. 5, 633 644. Blake, J., 1981. Peptide segment coupling in aqueous medium: silver ion activation of the thiolcarboxyl group. Int. J. Pept. Protein Res. 17, 273 274. Boyd, B., Richardson, S., Gariepy, J., 1991. Serological responses to the B subunit of Shiga-like toxin 1 and its peptide fragments indicate that the B subunit is a vaccine candidate to counter action of the toxin. Infect. Immun. 59, 750 757. Briand, J.P., Andre, C., Tuaillon, N., Herve, L., Neimark, J., Muller, S., 1992. Multiple autoepitope presentation for specic detection of antibodies in primary biliary cirrhosis. Hepatology 16, 1395 1403. Brown, F., 1992. New approaches to vaccination against footand-mouth disease. Vaccine 10, 1022 1026. Butz, S., Rawer, S., Rapp, W., Birsner, U., 1994. Immunization and afnity purication of antibodies using resin-immobilized lysine-branched synthetic peptides. Pept. Res. 7, 20 23. Bycroft, B.W., Chane, W.C., Chhabra, S.R., Hone, N.D., 1993. A novel lysine-protecting procedure for continuous-ow solid-phase synthesis of branched peptides. J. Chem. Soc. Chem. Commun., 778 779. Callaway, C., Seryshev, A., Wang, J.P. et al., 1994. Localization of the high and low afnity w 3 Hxryanodine binding sites on the skeletal muscle Ca2q release channel. J. Biol. Chem. 269, 15876 15884. Calvo-Calle, J.M., Nardin, E.H., Clavijo, P. et al., 1992. Recognition of different domains of the Plasmodium falciparum CS protein by the sera of naturally infected individuals compared with those of sporozoite-immunized volunteers. J. Immunol. 149, 2695 2701. Calvo-Calle, J.M., de Oliveira, G.A., Clavijo, P. et al., 1993. Immunogenicity of multiple antigen peptides containing B and non-repeat T cell epitopes of the circumsporozoite protein of Plasmodium falciparum. J. Immunol. 150, 1403 1412. Caponi, L., Pegoraro, S., Di Bartolo, V., Rovero, P., Revoltella, R., Bombardieri, S., 1995. Autoantibodies directed
K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229 cell epitope of the heat shock protein PfHsp70-1 antigen of Plasmodium falciparum using a multiple antigenic peptide. Parasite Immunol. 22, 535 543. DeLong, R., Stephenson, K., Loftus, T. et al., 1997. Characterization of complexes of oligonucleotides with polyamidoamine starburst dendrimers and effects on intracellular delivery. J. Pharmacol. Sci. 86, 762 764. De Oliveira, G.A., Clavijo, P., Nussenzweig, R.S., Nardin, E.H., 1994. Immunogenicity of an alum-adsorbed synthetic multiple-antigen peptide based on B- and T-cell epitopes of the Plasmodium falciparum CS protein: possible vaccine application. Vaccine 12, 1012 1017. De Santis, C., Lopalco, L., Robbioni, P. et al., 1994. Human antibodies to immunodominant C5 region of HIV-1 gp120 cross-react with HLA class I on activated cells. AIDS Res. Hum. Retroviruses 10, 157 162. Defoort, J.P., Nardelli, B., Huang, W., Ho, D.D., Tam, J.P., 1992a. Macromolecular assemblage in the design of a synthetic AIDS vaccine. Proc. Natl. Acad. Sci. USA 89, 3879 3883. Defoort, J.P., Nardelli, B., Huang, W., Tam, J.P., 1992b. A rational design of synthetic peptide vaccine with a built-in adjuvant. A modular approach for unambiguity. Int. J. Pept. Protein Res. 40, 214 221. Del Giudice, G., Tougne, C., Louis, J.A. et al., 1990. A multiple antigen peptide from the repetitive sequence of the Plasmodium malariae circumsporozoite protein induces a specic antibody response in mice of various H-2 haplotypes. Eur. J. Immunol. 20, 1619 1622. Del Guercio, M.F., Alexander, J., Kubo, R.T. et al., 1997. Potent immunogenic short linear peptide constructs composed of B cell epitopes and Pan DR T helper epitopes PADRE. for antibody responses in vivo. Vaccine 15, 441 448. Demotz, S., Danieli, C., Wallny, H.J., Majdic, O., 1994. Inhibition of peptide binding to DR molecules by a leupeptin-induced invariant chain fragment. Mol. Immunol. 31, 885 893. Digweed, M., Gunthert, U., Schneider, R., Seyschab, H., Friedl, R., Sperling, K., 1995. Irreversible repression of DNA synthesis in Fanconi anemia cells is alleviated by the product of a novel cyclin-related gene. Mol. Cell Biol. 15, 305 314. Drijfhout, J.W., Bloemhoff, W., 1991. A new synthetic functionalized antigen carrier. Int. J. Pept. Prot. Res. 37, 27 32. Duriez, P.J., Desnoyers, S., Hoack, J.C. et al., 1997. Characterization of anti-peptide antibodies directed towards the automodication domain and apoptotic fragment of poly ADP-ribose. polymerase. Biochim. Biophys. Acta 1334, 65 72. Edwards, R.J., Singleton, A.M., Murray, B.P., Murray, S., Boobis, A.R., Davies, D.S., 1991. Identication of a functionally conserved surface region of rat cytochromes P450IA. Biochem. J. 278, 749 757. Eicheler, W., Tuohimaa, P., Vilja, P., Adermann, K., Forssmann, W.G., Aumuller, G., 1994. Immunocytochemical lo-
221
calization of human 5 alpha-reductase 2 with polyclonal antibodies in androgen target and non-target human tissues. J. Histochem. Cytochem. 42, 667 675. Emery-Goodman, A., Hirling, H., Scarpellino, L., Henderson, B., Kuhn, L.C., 1993. Iron regulatory factor expressed from recombinant baculovirus: conversion between the RNA-binding apoprotein and Fe S cluster containing aconitase. Nucleic Acids Res. 21, 1457 1461. Esfand, R., Tomalia, D.A., 2001. Polyamidoamine. PAMAM. dendrimers: from biomimicry to drug delivery and biomedical applications. Drug Discov. Today 6, 427 436. Estaquier, J., Boutillon, C., Ameisen, J.C. et al., 1992. Determination of B-cell epitopes of nef HIV-I protein: immunogenicity related to their structure. Mol. Immunol. 29, 1337 1345. Estaquier, J., Boutillon, C., Gras-Masse, H. et al., 1993. Comprehensive delineation of antigenic and immunogenic properties of peptides derived from the nef HIV-1 regulatory protein. Vaccine 11, 1083 1092. Fairlie, D.P., West, M.L., Wong, A.K., 1998. Towards protein surface mimetics. Curr. Med. Chem. 5, 29 62. Farris, A.D., Brown, L., Reynolds, P. et al., 1999. Induction of autoimmunity by multivalent immunodominant and subdominant T cell determinants of La SS-B.. J. Immunol. 162, 3079 3087. Fassina, G., 1992. Oriented immobilization of peptide ligands on solid supports. J. Chromatogr. 591, 99 106. Fassina, G., Cassani, G., 1993. Peptide-based assay for the identication of endothelin-converting enzyme inhibitors. Pept. Res. 6, 73 78. Fassina, G., Corti, A., Cassani, G., 1992a. Afnity enhancement of complementary peptide recognition. Int. J. Pept. Protein Res. 39, 549 556. Fassina, G., Cassani, G., Corti, A., 1992b. Binding of human tumor necrosis factor alpha to multimeric complementary peptides. Arch. Biochem. Biophys. 296, 137 143. Felici, F., Luzzago, A., Folgori, A., Cortese, R., 1993. Mimicking of discontinuous epitopes by phage-displayed peptides, II. Selection of clones recognized by a protective monoclonal antibody against the Bordetella pertussis toxin from phage peptide libraries. Gene 128, 21 27. Ferru, I., Georges, B., Bossus, M. et al., 1997. Analysis of the immune response elicited by a multiple antigen peptide MAP. composed of two distinct protective antigens derived from the parasite Schistosoma mansoni. Parasite Immunol. 19, 1 11. Fields, C.G., Mickelson, D.J., Drake, S.L., McCarthy, J.B., Fields, G.B., 1993. Melanoma cell adhesion and spreading activities of a synthetic 124-residue triple-helical mini-collagen. J. Biol. Chem. 268, 14153 14160. Finkel, T., Theriot, J.A., Dise, K.R., Tomaselli, G.F., Goldschmidt-Clermont, P.J., 1994. Dynamic actin structures stabilized by prolin. Proc. Natl. Acad. Sci. USA 91, 1510 1514. Florence, A.T., Sakthivel, T., Toth, I., 2000. Oral uptake and
222
K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229 aconitase. A functional assay for translational repression and direct demonstration of the iron switch. Eur. J. Biochem. 218, 657 667. Grillot, D., Valmori, D., Lambert, P.H., Corradin, G., Del Giudice, G., 1993. Presentation of T-cell epitopes assembled as multiple-antigen peptides to murine and human T lymphocytes. Infect. Immun. 61, 3064 3067. Habluetzel, A., Pessi, A., Bianchi, E., Rotigliano, G., Esposito, F., 1991. Multiple antigen peptides for specic detection of antibodies to a malaria antigen in human sera. Immunol. Lett. 30, 75 80. Haensler, J., Szoka Jr., F.C., 1993. Polyamidoamine cascade polymers mediate efcient transfection of cells in culture. Bioconjug. Chem. 4, 372 379. Hahn, K.W., Klis, W.A., Stewart, J.M., 1990. Design and synthesis of a peptide having chymotrypsin-like esterase activity. Science 248, 1544 1547. Halimi, H., Rivaille, P., 1993. Immune response related to the molecular structure of a peptide from the cholera toxin B subunit. Vaccine 11, 1233 1239. Helling, F., Shang, A., Calves, M. et al., 1994. GD3 vaccines for melanoma: superior immunogenicity of keyhole limpet hemocyanin conjugate vaccines. Cancer Res. 54, 197 203. Henderson, B.R., Seiser, C., Kuhn, L.C., 1993. Characterization of a second RNA-binding protein in rodents with specicity for iron-responsive elements. J. Biol. Chem. 268, 27327 27334. Hojo, H., Aimoto, S., 1992. Protein-synthesis using s-alkyl thioester of partially protected peptide segments synthesis of DNA-binding protein of Bacillus-stearothermophilus. Bull. Chem. Soc. Jpn. 64, 3055 3063. Hsu, S.C., Chargelegue, D., Obeid, O.E., Steward, M.W., 1999. Synergistic effect of immunization with a peptide cocktail inducing antibody, helper and cytotoxic T-cell responses on protection against respiratory syncytial virus. J. Gen. Virol. 80, 1401 1405. Huang, W., Nardelli, B., Tam, J.P., 1994. Lipophilic multiple antigen peptide system for peptide immunogen and synthetic vaccine. Mol. Immunol. 31, 1191 1199. Iio, A., Mochii, M., Agata, K., Kodoma, R., Eguchi, G., 1994. Expression of the retinal pigmented epithelial cell-specic pp344 gene during development of the chicken eye and identication of its product. Dev. Growth Differ. 36, 155 164. Ingham, K.C., Brew, S.A., Migliorini, M., 1994. An unusual heparin-binding peptide from the carboxy-terminal hep-2 region of bronectin. Arch. Biochem. Biophys. 314, 242 246. Itoh, N., Jobo, K., Tsujimoto, K., Ohta, M., Kawasaki, T., 1993. Two truncated forms of rat insulin receptor-related receptor. J. Biol. Chem. 268, 17983 17986. Iwamoto, Y., Nomizu, M., Yamada, Y., Ito, Y., Tanaka, K., Sugioka, Y., 1996. Inhibition of angiogenesis, tumour growth and experimental metastasis of human brosarcoma cells HT1080 by a multimeric form of the laminin sequence Tyr Ile Gly Ser Arg YIGSR.. Br. J. Cancer 73, 589 595. Jackwood, M.W., Hilt, D.A., 1995. Production and immuno-
translocation of a polylysine dendrimer with a lipid surface. J. Control Release 65, 253 259. Fraisier, C., Ebersold, A., Blomberg, J., Desgranges, C., 1994. Primary in vitro immunization with multimeric synthetic peptides of HIV-1 envelope glycoproteins: generation of neutralizing human monoclonal antibodies. J. Immunol. Methods 176, 9 22. Francis, M.J., Hastings, G.Z., Brown, F., McDermed, J., Lu, Y.-A., Tam, J.P., 1991. Immunological evaluation of the multiple antigen peptide MAP. system using the major immunogenic site of foot-and-mouth disease virus. Immunology 73, 249 254. Franke, E.D., Sette, A., Sacci Jr., J., Southwood, S., Corradin, G., Hoffman, S.L., 2000. A subdominant CD8q. cytotoxic T lymphocyte CTL. epitope from the Plasmodium yoelii circumsporozoite protein induces CTLs that eliminate infected hepatocytes from culture. Infect. Immun. 68, 3403 3411. Frey, H., 1998. From random coil to extended nanocylinder: dendrimer fragments shape polymer chains. Angew. Chem. Int. Ed. 37, 2193 2197. Gaertner, H.F., Rose, K., Cotton, R., Timms, D., Camble, R., Offord, R.E., 1992. Construction of protein analogues by site-specic condensation of unprotected fragments. Bioconj. Chem. 3, 262 268. Genco, C.A., Odusanya, B.M., Potempa, J., MikolajczykPawlinska, J., Travis, J., 1998. A peptide domain on gingipain R which confers immunity against Porphyromonas gingi alis infection in mice. Infect. Immun. 66, 4108 4114. Gettemans, J., De Ville, Y., Vandekerckhove, J., Waelkens, E., 1993. Purication and partial amino acid sequence of the actin fragmin kinase from Physarum polycephalum. Eur. J. Biochem. 214, 111 119. Ghadiri, M.R., Case, M.A., 1993. De-novo design of a novel heterodinuclear 3-helix bundle metalloprotein. Angew. Chem. Int. Ed. Engl. 32, 1594 1597. Gibbons, W.A., Hughes, R.A., Szeto, A., et al., 1990. Lipidic peptides.1. Synthesis, resolution and structural elucidation of lipidic amino-acids and their homo-oligomers and heterooligomers. Liebigs Ann. Chem. 1175 1183. Godard, I., Estaquier, J., Zenner, L. et al., 1994. Antigenicity and immunogenicity of P30-derived peptides in experimental models of toxoplasmosis. Mol. Immunol. 31, 1353 1363. Gomara, M.J., Ercilla, G., Alsina, M.A., Haro, I., 2000a. Assessment of synthetic peptides for hepatitis A diagnosis using biosensor technology. J. Immunol. Methods 246, 13 24. Gomara, M.J., Riedemann, S., Vega, I., Ibarra, H., Ercilla, G., Haro, I., 2000b. Use of linear and multiple antigenic peptides in the immunodiagnosis of acute hepatitis A virus infection. J. Immunol. Methods 234, 23 34. Goodman, M., Bhumralkar, M., Jefferson, E.A., Kwak, J., Locardi, E., 1998. Collagen mimetics. Biopolymers 47, 127 142. Gray, N.K., Quick, S., Goossen, B. et al., 1993. Recombinant iron-regulatory factor functions as an iron-responsive-element-binding protein, a translational repressor and an
K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229 genicity of multiple antigenic peptide MAP. constructs derived from the S1 glycoprotein of infectious bronchitis virus IBV.. Adv. Exp. Med. Biol. 380, 213 219. James, J.A., Harley, J.B., 1998. A model of peptide-induced lupus autoimmune B cell epitope spreading is strain specic and is not H-2 restricted in mice. J. Immunol. 160, 502 508. James, J.A., Gross, T., Scoeld, R.H., Harley, J.B., 1995. Immunoglobulin epitope spreading and autoimmune disease after peptide immunization: Sm BrB -derived PPPGMRPP and PPPGIRGP induce spliceosome autoimmunity. J. Exp. Med. 181, 453 461. Jaso-Friedmann, L., Leary 3rd, J.H., Weisman, Z., Evans, D.L., 1996. Activation of nonspecic cytotoxic cells with a multiple antigenic peptide: specicity and requirements for receptor crosslinkage. Cell Immunol. 170, 195 201. Jefferson, E.A., Locardi, E., Goodman, M., 1998. Incorporation of achiral peptoid-based trimeric sequences into collagen mimetics. J. Am. Chem. Soc. 120, 7420 7428. Kabeya, H., Ohashi, K., Ohishi, K., Sugimoto, C., Amanuma, H., Onuma, M., 1996. An effective peptide vaccine to eliminate bovine leukaemia virus BLV. infected cells in carrier sheep. Vaccine 14, 1118 1122. Kajiwara, H., Tomooka, N., 1998. Comparative analysis of genus Vigna seeds using antiserum against a synthesized multiple antigenic peptide. Electrophoresis 19, 3110 3113. Kamo, K., Jordan, R., Hsu, H., Hudson, D., 1992. Development of a monoclonal antibody to the conserved region of p34cdc2 protein kinase. J. Immunol. Methods 156, 163 170. Kaneko, Y., Okada, N., Baranyi, L., Azuma, T., Okada, H., 1995. Antagonistic peptides against human anaphylatoxin C5a. Immunology 86, 149 154. Kang, D., Okubo, K., Hamasaki, N., Kuroda, N., Shiraki, H., 1992. A structural study of the membrane domain of band 3 by tryptic digestion. Conformational change of band 3 in situ induced by alkali treatment. J. Biol. Chem. 267, 19211 19217. Kates, S.A., Daniels, S.B., Albericio, F., 1993. Automated allyl cleavage for continuous-ow synthesis of cyclic and branched peptides. Anal. Biochem. 212, 303 310. Kawase, M., Kurikawa, N., Miura, N. et al., 2000. Immobilization of ligand-modied polyamidoamine dendrimer for cultivation of hepatoma cells. Articial Organs 24, 18 22. Keller, O., Rudinger, J., 1975. Preparation and some properties of maleimido acids and maleoyl derivatives of peptides. Helv. Chim. Acta 58, 531 534. Kelker, H.C., Schlesinger, D., Valentine, F.T., 1994. Immunogenic and antigenic properties of an HIV-1 gp120-derived multiple chain peptide. J. Immunol. 152, 4139 4148. Kemp, D.S., Petrakis, K.S., 1981. Synthesis and conformational-analysis of cis,cis-1,3,5-trimethylcyclohexane-1,3,5-tricarboxylic acid. J. Org. Chem. 46, 5140 5143. Khalil, M., Inaba, K., Steinman, R., Ravetch, J., Diamond, B., 2001. T cell studies in a peptide-induced model of systemic lupus erythematosus. J. Immunol. 166, 1667 1674. Khan, C.M., Villarreal-Ramos, B., Pierce, R.J. et al., 1994. Construction, expression, and immunogenicity of multiple
223
tandem copies of the Schistosoma mansoni peptide 115-131 of the P28 glutathione S-transferase expressed as C-terminal fusions to tetanus toxin fragment C in a live aro-attenuated vaccine strain of Salmonella. J. Immunol. 153, 5634 5642. Kim, Y., Zimmerman, S.C., 1998. Applications of dendrimers in bio-organic chemistry. Curr. Opin. Chem. Biol. 2, 733 742. Kim, W.H., Schnaper, H.W., Nomizu, M., Yamada, Y., Kleinman, H.K., 1994. Apoptosis in human brosarcoma cells is induced by a multimeric synthetic Tyr Ile Gly Ser Arg YIGSR.-containing polypeptide from laminin. Cancer Res. 54, 5005 5010. King, T.P., Li, Y., Kochoumian, L., 1978. Preparation of protein conjugates via intermolecular disulde bond formation. Biochemistry 17, 1499 1506. King, T.P., Zhao, S.W., Lam, T., 1986. Preparation of protein conjugates via intermolecular hydrazone linkage. Biochemstry 25, 5774. Kojima, C., Kono, K., Maruyama, K., Takagishi, T., 2000. Synthesis of polyamidoamine dendrimers having polyethylene glycol. grafts and their ability to encapsulate anticancer drugs. Bioconjugate Chem. 11, 910 917. Kuhn, M., Kulaksiz, H., Adermann, K., Rechkemmer, G., Forssmann, W.G., 1994. Radioimmunoassay for circulating human guanylin. FEBS Lett. 341, 218 222. Kukowska-Latallo, J.F., Bielinska, A.U., Johnson, J., Spindler, R., Tomalia, D.A., Baker Jr., J.R., 1996. Efcient transfer of genetic material into mammalian cells using starburst polyamidoamine dendrimers. Proc. Natl. Acad. Sci. USA 93, 4897 4902. Kutty, R.K., Nagineni, C.N., Kutty, G., Hooks, J.J., Chader, G.J., Wiggert, B., 1994. Increased expression of heme oxygenase-1 in human retinal pigment epithelial cells by transforming growth factor-beta. J. Cell Physiol. 159, 371 378. Lankinen, H., Telford, E., MacDonald, D., Marsden, H., 1989. The unique N-terminal domain of the large subunit of herpes simplex virus ribonucleotide reductase is preferentially sensitive to proteolysis. J. Gen. Virol. 70, 3159 3169. Le, T.P., Church, L.W., Corradin, G. et al., 1998. Immunogenicity of Plasmodium falciparum circumsporozoite protein multiple antigen peptide vaccine formulated with different adjuvants. Vaccine 16, 305 312. Levi, M., Ruden, U., Birx, D. et al., 1993. Effects of adjuvants and multiple antigen peptides on humoral and cellular immune responses to gp160 of HIV-1. J. Acquir. Immune Dec. Syndr. 6, 855 864. Li, J.K., Yang, Y.Y., 1990. Mapping of two immunodominant antigenic epitopes conserved among the major inner capsid protein, VP7 of ve bluetongue viruses. Virology 178, 552 559. Liew, F.Y., Millott, S.M., Schmidt, J.A., 1990. A repetitive peptide of Leishmania can activate T helper type 2 cells and enhance disease progression. J. Exp. Med. 172, 1359 1365. Lingappa, J.R., Martin, R.L., Wong, M.L., Ganem, D., Welch, W.J., Lingappa, V.R., 1994. A eukaryotic cytosolic chaper-
224
K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229 cally dened constructs of a peptide. Mol. Immunol. 29, 793 800. Marsden, H.S., Owsianka, A.M., Graham, S., McLean, G.W., Robertson, C.A., Subak-Sharpe, J.H., 1992. Advantages of branched peptides in serodiagnosis. Detection of HIVspecic antibodies and the use of glycine spacers to increase sensitivity. J. Immunol. Methods 147, 65 72. Marsden, H.S., Murphy, M., McVey, G.L., MacEachran, K.A., Owsianka, A.M., Stow, N.D., 1994. Role of the carboxy terminus of herpes simplex virus type 1 DNA polymerase in its interaction with UL42. J. Gen. Virol. 75, 3127 3135. Marussig, M., Renia, L., Motard, A. et al., 1997. Linear and multiple antigen peptides containing dened T and B epitopes of the Plasmodium yoelii circumsporozoite protein: antibody-mediated protection and boosting by sporozoite infection. Int. Immunol. 9, 1817 1824. Mason, L.J., Timothy, L.M., Isenberg, D.A., Kalsi, J.K., 1999. Immunization with a peptide of Sm BrB results in limited epitope spreading but not autoimmune disease. J. Immunol. 162, 5099 5105. McLean, G., Rixon, F., Langeland, N., Haarr, L., Marsden, H., 1990. Identication and characterization of the virion protein products of herpes simplex virus type 1 gene UL47. J. Gen. Virol. 71, 2953 2960. McLean, G.W., Owsianka, A.M., Subak-Sharpe, J.H., Marsden, H.S., 1991. Generation of anti-peptide and anti-protein sera. Effect of peptide presentation on immunogenicity. J. Immunol. Methods 137, 149 157. McLean, G.W., Cross, A.M., Munns, M.S., Marsden, H.S., 1992. Rapid attachment of a helper T cell epitope to branched peptides by fragment condensation to give enhanced immunogenicity. J. Immunol. Methods 155, 113 120. McNeil, H.P., Frenkel, D.P., Austen, K.F., Friend, D.S., Stevens, R.L., 1992. Translation and granule localization of mouse mast cell protease-5. Immunodetection with specic antipeptide Ig. J. Immunol. 149, 2466 2472. Merrield, R.B., 1963. Solid phase peptide synthesis. I. The synthesis of a tetrapeptide. J. Am. Chem. Soc. 85, 2149 2153. Meziere, C., Stockl, F., Batsford, S., Vogt, A., Muller, S., 1994. Antibodies to DNA, chromatin core particles and histones in mice with graft-versus-host disease and their involvement in glomerular injury. Clin. Exp. Immunol. 98, 287 294. Migliorini, P., Betschart, B., Corradin, G., 1993. Malaria vaccine: immunization of mice with a synthetic T cell helper epitope alone leads to protective immunity. Eur J. Immunol. 23, 582 585. Miotto, G., Venerando, R., Marin, O., Siliprandi, N., Mortimore, G.E., 1994. Inhibition of macroautophagy and proteolysis in the isolated rat hepatocyte by a nontransportable derivative of the multiple antigen peptide Leu8 Lys4 Lys2 Lys beta Ala. J. Biol. Chem. 269, 25348 25353. Molnar, E., Baude, A., Richmond, S.A., Patel, P.B., Somogyi, P., McIlhinney, R.A., 1993. Biochemical and immunocytochemical characterization of antipeptide antibodies to a cloned GluR1 glutamate receptor subunit: cellular and
onin is associated with a high molecular weight intermediate in the assembly of hepatitis B virus capsid, a multimeric particle. J. Cell Biol. 125, 99 111. Liu, C.-F., Tam, J.P., 1994. Peptide segment ligation strategy without use of protecting groups. Proc. Natl. Acad. Sci. USA 91, 6584 6588. Liu, C.-F., Tam, J.P., 1997. Synthesis of a symmetric peptide dendrimer. Assembly of a cyclic peptide on a small tetraacetate template. Chem. Commun. 1619 1620. Liu, C.-F., Rao, C., Tam, J.P., 1996. Orthogonal ligation of unprotected peptide segments through pseudoproline formation for the synthesis of HIV-1 protease analogs. J. Am. Chem. Soc. 118, 307 312. Liu, C.-F., Rao, C., Tam, J.P., 1999. Use of orthogonal ligation methods for the synthesis of a hetero peptide dendrimer. In: Fields, G.B., Tam, J.P., Barany, G. Eds.., Proceedings of the 16th American Peptide Symposium. Kluwer Academic Publishers, London, pp. 118 119. Lo-Man, R., Martineau, P., Betton, J.M., Hofnung, M., Leclerc, C., 1994. Molecular context of a viral T cell determinant within a chimeric bacterial protein alters the diversity of its T cell recognition. J. Immunol. 152, 5660 5669. Lu, Y.-A., Clavijo, P., Galantino, M., Shen, Z.Y., Liu, W., Tam, J.P., 1991. Chemically unambiguous peptide immunogen: preparation, orientation and antigenicity of puried peptide conjugated to the multiple antigen peptide system. Mol. Immunol. 28, 623 630. Lu, Y.-A., Yang, J.-L., Tam, J.P., 2001. Functional and structural comparisons of linear and dendritic arrays of polypeptides. Proceedings of 17th American Peptide Symposium, in press. Lugovskoi, A.A., Rybakov, S.S., Ivaniushchenkov, V.N. et al., 1992. Protection of naturally susceptible animals against foot-and-mouth disease with a peptide, synthesized on a lysine matrix. Bioorg. Khim. 18, 942 950. Malouf, N.N., McMahon, D.K., Hainsworth, C.N., Kay, B.K., 1992. A two-motif isoform of the major calcium channel subunit in skeletal muscle. Neuron 8, 899 906. Mammen, M., Choi, S.-K., Whitesides, G.M., 1998. Polyvalent interactions in biological systems: implications for design and use of multivalent ligands and inhibitors. Angew. Chem. Int. Ed. 37, 2754 2794. Manivel, V., Tripathy, A., Durgapal, H., Kumar, A., Panda, S.K., Rao, K.V., 1993. A self-associating hepatitis B surface antigen-derived peptide that is immunogenic in alum. Vaccine 11, 366 371. Manki, A., Ono, T., Uenaka, A., Seino, Y., Nakayama, E., 1998. Vaccination with multiple antigen peptide as rejection antigen peptide in murine leukemia. Cancer Res. 58, 1960 1964. Marchini, B., Dolcher, M.P., Sabbatini, A., Klein, G., Migliorini, P., 1994. Immune response to different sequences of the EBNA I molecule in Epstein-Barr virus-related disorders and in autoimmune diseases. J. Autoimmun. 7, 179 191. Marguerite, M., Bossus, M., Mazingue, C. et al., 1992. Analysis of antigenicity and immunogenicity of ve different chemi-
K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229 subcellular distribution in the rat forebrain. Neuroscience 53, 307 326. Monsalve, R.I., Gonzalez de la Pena, M.A., Menendez-Arias, L., Lopez-Otin, C., Villalba, M., Rodriguez, R., 1993. Characterization of a new oriental-mustard Brassica juncea. allergen, Bra j IE: detection of an allergenic epitope. Biochem. J. 293, 625 632. Moradi-Ameli, M., Rousseau, J.C., Kleman, J.P. et al., 1994. Diversity in the processing events at the N-terminus of type-V collagen. Eur. J. Biochem. 221, 987 995. Moreno, C.A., Rodriguez, R., Oliveira, G.A. et al., 1999. Preclinical evaluation of a synthetic Plasmodium falciparum MAP malaria vaccine in Aotus monkeys and mice. Vaccine 18, 89 99. Mortimore, G.E., Wert Jr., J.J., Miotto, G., Venerando, R., Kadowaki, M., 1994. Leucine-specic binding of photoreactive Leu7-MAP to a high molecular weight protein on the plasma membrane of the isolated rat hepatocyte. Biochem. Biophys. Res. Commun. 203, 200 208. Muir, T.W., Williams, M.J., Ginsberg, M.H., Kent, S.B.H., 1994. Design and chemical synthesis of a neoprotein structural model for the cytoplasmic domain of a multisubunit cell-surface receptor: integrin alpha IIb beta 3 platelet GPIIb IIIa.. Biochemistry 33, 7701 7708. Munesinghe, D.Y., Clavijo, P., Calle, M.C., Nussenzweig, R.S., Nardin, E., 1991. Immunogenicity of multiple antigen peptides MAP. containing T and B cell epitopes of the repeat region of the P. falciparum circumsporozoite protein. Eur. J. Immunol. 21, 3015 3120. Murer, P., Seebach, D., 1995. Synthesis and properties of rst-generation to 3rd-generation dendrimers with doubly and triply branched chiral building-blocks. Angew. Chem. Int. Ed. Engl. 34, 2116 2119. Nagamatsu, S., Sawa, H., Kamada, K., Nakamichi, Y., Yoshimoto, K., Hoshino, T., 1993. Neuron-specic glucose transporter NSGT.: CNS distribution of GLUT3 rat glucose transporter RGT3. in rat central neurons. FEBS Lett. 334, 289 295. Nakitare, G.W., Elliott, R.M., 1993. Expression of the Bunyamwera virus M genome segment and intracellular localization of NSm. Virology 195, 511 520. Nara, K., Ito, S., Ito, T. et al., 1994. Elastase inhibitor elan is a new type of proteinase inhibitor which has a transglutaminase-mediated anchoring sequence termed cementoin. J. Biochem. 115, 441 448. Nardelli, B., Tam, J.P., 1993. Cellular immune responses induced by in vivo priming with a lipid-conjugated multimeric antigen peptide. Immunology 79, 355 361. Nardelli, B., Defoort, J.P., Huang, W., Tam, J.P., 1992a. Design of a complete synthetic peptide-based AIDS vaccine with a built-in adjuvant. AIDS Res. Hum. Retroviruses 8, 1405 1407. Nardelli, B., Lu, Y.-A., Shiu, D.R., Delpierre-Defoort, C., Profy, A.T., Tam, J.P., 1992b. A chemically dened synthetic vaccine model for HIV-1. J. Immunol. 148, 914 920. Nardelli, B., Haser, P.B., Tam, J.P., 1994. Oral administration of an antigenic synthetic lipopeptide MAP-P3C. evokes
225
salivary antibodies and systemic humoral and cellular responses. Vaccine 12, 1335 1339. Nardin, E.H., Nussenzweig, R.S., 1993. T cell responses to pre-erythrocytic stages of malaria: role in protection and vaccine development against pre-erythrocytic stages. Annu. Rev. Immunol. 11, 687 727. Nardin, E.H., Calvo-Calle, J.M., Oliveira, G.A. et al., 1998. Plasmodium falciparum polyoximes: highly immunogenic synthetic vaccines constructed by chemoselective ligation of repeat B-cell epitopes and a universal T-cell epitope of CS protein. Vaccine 16, 590 600. Nardin, E.H., Calvo-Calle, J.M., Oliveira, G.A. et al., 2001. A totally synthetic polyoxime malaria vaccine containing Plasmodium falciparum B cell and universal T cell epitopes elicits immune responses in volunteers of diverse HLA types. J. Immunol. 166, 481 489. Naruse, H., Ogasawara, K., Kaneda, R. et al., 1994. A potential peptide vaccine against two different strains of inuenza virus isolated at intervals of about 10 years. Proc. Natl. Acad. Sci. USA 91, 9588 9592. Newkome, G.R., Lin, X., Weis, C.D., 1991. Polytryptophane terminated dendritic macromolecules. Tetrahedron Asymmetry 2, 957 960. Nieto, A., de la Luna, S., Barcena, J. et al., 1992. Nuclear transport of inuenza virus polymerase PA protein. Virus Res. 24, 65 75. Nomizu, M., Yamamura, K., Kleinman, H.K., Yamada, Y., 1993. Multimeric forms of Tyr Ile Gly Ser Arg YIGSR. peptide enhance the inhibition of tumor growth and metastasis. Cancer Res. 53, 3459 3461. Olszewska, W., Partidos, C.D., Steward, M.W., 2000a. Antipeptide antibody responses following intranasal immunization: effectiveness of mucosal adjuvants. Infect. Immun. 68, 4923 4929. Olszewska, W., Obeid, O.E., Steward, M.W., 2000b. Protection against measles virus-induced encephalitis by anti-mimotope antibodies: the role of antibody afnity. Virology 272, 98 105. ORourke, K.I., Huff, T.P., Leathers, C.W., Robinson, M.M., Gorham, J.R., 1994. SCID mouse spleen does not support scrapie agent replication. J. Gen. Virol. 75, 1511 1514. Ottl, J., Gabriel, D., Muller, H.Y.C.D., Musiol, H.-J., Bode, W., Moroder, L., 1999. Synthesis of disulde-bridged heterotrimeric collagen peptides. Conformational properties and digestion by matrix metalloproteinese. In: Shimonishi, Y. Ed.., Peptide Science Present and Future. Kluwer Academic Publishers, The Netherlands, pp. 485 489. Pallin, T.D., Tam, J.P., 1995. Cyclization of totally unprotected peptides in aqueous-solution by oxime formation. J. Chem. Soc. Chem. Commun. 19, 2021 2022. Pallin, T.D., Tam, J.P., 1996. Assembly of cyclic peptide dendrimers from unprotected linear building blocks in aqueous solution. Chem. Commun., 1345 1346. Pancre, V., Wolowczuk, I., Bossus, M. et al., 1994. Evaluation of the effect of Sm28GST-derived peptides in murine hepatosplenic schistosomiasis: interest of the lipopeptidic form of the C-terminal peptide. Mol. Immunol. 31, 1247 1256.
226
K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229 Roth, W., Heidemann, E., 1980. Triple helix-coil transition of covalently bridged collagen-like peptides. Biopolymers 19, 1909 1917. Sa, G., Murugesan, G., Jaye, M., Ivashchenko, Y., Fox, P.L., 1995. Activation of cytosolic phospholipase A2 by basic broblast growth factor via a p42 mitogen-activated protein kinase-dependent phosphorylation pathway in endothelial cells. J. Biol. Chem. 270, 2360 2366. Sabbatini, A., Bombardieri, S., Migliorini, P., 1993a. Autoantibodies from patients with systemic lupus erythematosus bind a shared sequence of SmD and Epstein-Barr virus-encoded nuclear antigen EBNA I. Eur. J. Immunol. 23, 1146 1152. Sabbatini, A., Dolcher, M.P., Marchini, B., Bombardieri, S., Migliorini, P., 1993b. Mapping of epitopes on the SmD molecule: the use of multiple antigen peptides to measure autoantibodies in systemic lupus erythematosus. J. Rheumatol. 20, 1679 1683. Saikawa, T., Anderson, S., Momoeda, M., Kajigaya, S., Young, N.S., 1993. Neutralizing linear epitopes of B19 parvovirus cluster in the VP1 unique and VP1 VP2 junction regions. J. Virol. 67, 3004 3009. Sakakibara, S., 1995. Synthesis of large peptides in solution. Biopolymers 37, 17 28. Sakamoto, M., Kamachi, T., Okura, I., Ueno, A., Mihara, H., 2001a. Photoinduced hydrogen evolution with peptide dendrimer-multi-ZnII.-porphyrin, viologen, and hydrogenase. Biopolymers 59, 103 109. Sakamoto, M., Ueno, A., Mihara, H., 2001b. Multipeptidemetalloporphyrin assembly on a dendrimer template and photoinduced electron transfer based on the dendrimer structure. Chemistry 7, 2449 2458. Sasaki, T., Kaiser, E.T., 1989. Helichrome-synthesis and enzymatic-activity of a designed hemeprotein. J. Am. Chem. Soc. 111, 380 381. Schacke, H., Schroder, H.C., Gamulin, V., Rinkevich, B., Muller, I.M., Muller, W.E., 1994. Molecular cloning of a tyrosine kinase gene from the marine sponge Geodia cydonium: a new member belonging to the receptor tyrosine kinase class II family. Mol. Membr. Biol. 11, 101 107. Schneider, J.P., Kelly, J.W., 1995. Templates that induce alpha-helical, beta-sheet, and loop conformations. Chem. Rev. 95, 2169 2187. Schott, M.E., Wells, D.T., Schlom, J., Abrams, S.I., 1996. Comparison of linear and branched peptide forms MAPs. in the induction of T helper responses to point-mutated ras immunogens. Cell Immunol. 174, 199 209. Schuck, P., 1997. Use of surface plasmon resonance to probe the equilibrium and dynamic aspects of interactions between biological macromolecules. Annu. Rev. Biophys. Biomol. Struct. 26, 541 566. Scoeld, R.H., Kurien, B.T., Zhang, F. et al., 1999. Protein protein interaction of the Ro-ribonucleoprotein particle using multiple antigenic peptides. Mol. Immunol. 36, 1093 1106.
Parry, M.E., Stow, N.D., Marsden, H.S., 1993. Purication and properties of the herpes simplex virus type 1 UL8 protein. J. Gen. Virol. 74, 607 612. Pegg, A.E., Wiest, L., Mummert, C., Dolan, M.E., 1991. Production of antibodies to peptide sequences present in human O6-alkylguanine-DNA alkyltransferase and their use to detect this protein in cell extracts. Carcinogenesis 12, 1671 1677. Perin, M.S., 1994. The COOH terminus of synaptotagmin mediates interaction with the neurexins. J. Biol. Chem. 269, 8576 8581. Pessi, A., Valmori, D., Migliorini, P. et al., 1991. Lack of H-2 restriction of the Plasmodium falciparum NANP. sequence as multiple antigen peptide. Eur. J. Immunol. 21, 2273 2276. Pinto, R.M., Gonzalez-Dankaart, J.F., Sanchez, G. et al., 1998. Enhancement of the immunogenicity of a synthetic peptide bearing a VP3 epitope of hepatitis A virus. FEBS Lett. 438, 106 110. Posnett, D.N., McGrath, H., Tam, J.P., 1988. A novel method for producing anti-peptide antibodies. Production of sitespecic antibodies to the T cell antigen receptor beta-chain. J. Biol. Chem. 263, 1719 1725. Purohit, G., Sakthivel, T., Florence, A.T., 2001. Interaction of cationic partial dendrimers with charged and neutral liposomes. Int. J. Pharm. 214, 71 76. Putterman, C., Diamond, B., 1998. Immunization with a peptide surrogate for double-stranded DNA dsDNA. induces autoantibody production and renal immunoglobulin deposition. J. Exp. Med. 188, 29 38. Rajagopalan, S., Heck, T.J., Iwamoto, T., Tomich, J.M., 1995. Use of the 3-nitro-2-pyridine sulfenyl protecting group to introduce N epsilon-branching at lysine during solid-phase peptide synthesis. I. Application to the synthesis of a peptide template containing two addressable sites. Int. J. Peptide Protein Res. 45, 173 179. Rao, C., Tam, J.P., 1994. Synthesis of peptide dendrimer. J. Am. Chem. Soc. 116, 6576 6975. Raymond, J.R., Olsen, C.L., Gettys, T.W., 1993. Cell-specic physical and functional coupling of human 5-HT1A receptors to inhibitory G protein alpha-subunits and lack of coupling to Gs alpha. Biochemistry 32, 11064 11073. Reynolds, S.R., Dahl, C.E., Harn, D.A., 1994. T and B epitope determination and analysis of multiple antigenic peptides for the Schistosoma mansoni experimental vaccine triosephosphate isomerase. J. Immunol. 152, 193 200. Rigby, M.A., Mackay, N., Reid, G., Osborne, R., Neil, J.C., Jarrett, O., 1996. Immunogenicity of a peptide from a major neutralising determinant of the feline immunodeciency virus surface glycoprotein. Vaccine 14, 1095 1102. Romero, P.J., Tam, J.P., Schlesinger, D. et al., 1988. Multiple T helper cell epitopes of the circumsporozoite protein of Plasmodium berghei. Eur. J. Immunol. 18, 1951 1957. Rose, K., 1994. Facile synthesis of homogeneous articial proteins. J. Am. Chem. Soc. 116, 30 33.
K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229 Shah, D.S., Sakthivel, T., Toth, I., Florence, A.T., Wilderspin, A.F., 2000. DNA transfection and transfected cell viability using amphipathic asymmetric dendrimers. Int. J. Pharm. 208, 41 48. Shao, J., Tam, J.P., 1995. Unprotected peptides as buildingblocks for the synthesis of peptide dendrimers with oxime, hydrazone, and thiazolidine linkages. J. Am. Chem. Soc. 117, 3893 3899. Sheldon, K., Liu, D., Ferguson, J., Gariepy, J., 1995. Loligomers: design of de novo peptide-based intracellular vehicles. Proc. Natl. Acad. Sci. USA 92, 2056 2060. Shi, Z., Buntel, C.J., Grifn, J.H., 1994. Isolation and characterization of the gene encoding 2,3-oxidosqualenelanosterol cyclase from Saccharomyces cere isiae. Proc. Natl. Acad. Sci. USA 91, 7370 7374. Shimizu, H., Ito, M., Miyahara, M. et al., 1994. Characterization of the myosin-binding subunit of smooth muscle myosin phosphatase. J. Biol. Chem. 269, 30407 30411. Shin, S.Y., Lee, M.K., Kim, S.Y., Jang, S.Y., Hahm, K.S., 1997. The use of multiple antigenic peptide MAP. in the immunodiagnosis of human immunodeciency virus infection. Biochem. Mol. Biol. Int. 43, 713 721. Sil, P., Misono, K., Sen, S., 1993. Myotrophin in human cardiomyopathic heart. Circ. Res. 73, 98 108. Simmer, J.P., Hu, C.C., Lau, E.C., Sarte, P., Slavkin, H.C., Fincham, A.G., 1994. Alternative splicing of the mouse amelogenin primary RNA transcript. Calcif. Tissue Int. 55, 302 310. Simmonds, P., Rose, K.A., Graham, S. et al., 1993. Mapping of serotype-specic, immunodominant epitopes in the NS-4 region of hepatitis C virus HCV.: use of type-specic peptides to serologically differentiate infections with HCV types 1, 2, and 3. J. Clin. Microbiol. 31, 1493 1503. Sinclair, M.C., McLauchlan, J., Marsden, H., Brown, S.M., 1994. Characterization of a herpes simplex virus type 1 deletion variant 1703. which under-produces Vmw63 during immediate early conditions of infection. J. Gen. Virol. 75, 1083 1089. Singh, P., 1998. Terminal groups in starburst dendrimers: activation and reactions with proteins. Bioconjug. Chem. 9, 54 63. Sinnis, P., Clavijo, P., Fenyo, D., Chait, B.T., Cerami, C., Nussenzweig, V., 1994. Structural and functional properties of region II-plus of the malaria circumsporozoite protein. J. Exp. Med. 180, 297 306. Smith, D.J., Taubman, M.A., Holmberg, C.F., Eastcott, J., King, W.F., Ali-Salaam, P., 1993. Antigenicity and immunogenicity of a synthetic peptide derived from a glucan-binding domain of mutans streptococcal glucosyltransferase. Infect. Immun. 61, 2899 2905. Smith, D.J., Taubman, M.A., King, W.F., Eida, S., Powell, J.R., Eastcott, J., 1994a. Immunological characteristics of a synthetic peptide associated with a catalytic domain of mutans streptococcal glucosyltransferase. Infect. Immun. 62, 5470 5476.
227
Smith, M.A., Kutty, R.K., Richey, P.L. et al., 1994b. Heme oxygenase-1 is associated with the neurobrillary pathology of Alzheimers disease. Am. J. Pathol. 145, 42 47. Spetzler, J.C., Tam, J.P., 1995. Unprotected peptides as building blocks for branched peptides and peptide dendrimers. Int. J. Peptide Protein Res. 45, 78 85. Spetzler, J.C., Tam, J.P., 1996. Self-assembly of cyclic peptides on a dendrimer: multiple cyclic antigen peptides. Peptide Res. 9, 290. St John, L.C., Bell, F.P., Kezdy, F.J. et al., 1991. Production of an antibody to arterial fatty acid-binding protein FABP. using a synthetic peptide as the antigen. Comp. Biochem. Physiol. B. 99, 431 435. Stout, J.G., Brittsan, A., Kirley, T.L., 1994. Brain ECTO MgATPase is not the neural cell adhesion molecule. Biochem. Mol. Biol. Int. 33, 1091 1098. Suzuki, Y.S., Wang, Y., Takemoto, J.Y., 1992. Syringomycinstimulated phosphorylation of the plasma-membrane HqATPase from red beet storage tissue. Plant Physiol. 99, 1314 1320. Szymkowski, D.E., Deering, R.A., 1990. Identication and characterization of a Dictyostelium discoideum ribosomal protein gene. Nucleic Acids Res. 18, 4695 4701. Tam, J.P., 1988. Synthetic peptide vaccine design-synthesis and properties of a high-density multiple antigenic peptide system. Proc. Natl. Acad. Sci. USA 85, 5409 5413. Tam, J.P., 1996. Recent advances in multiple antigen peptides. J. Immunol. Methods 196, 17 32. Tam, J.P., 2000. Synthesis of peptides and peptidomimetics, Houben-Weyl Methods of Organic Chemistry. In: Goodman, M. Ed.., Peptide dendrimers and protein mimetics, Thieme, Stuttgart. Tam, J.P., Lu, Y.-A., 1989. Vaccine engineering-enhancement of immunogenicity of synthetic peptide vaccines related to hepatitis in chemically dened models consisting of t-cell and b-cell epitopes. Proc. Natl. Acad. Sci. USA 86, 9084 9088. Tam, J.P., Lu, Y.-A., 1995. Coupling difculty associated with interchain clustering and phase-transition in solid-phase peptide-synthesis. J. Am. Chem. Soc. 117, 12058 12063. Tam, J.P., Lu, Y.-A., 1998. A biomimetic strategy in the synthesis and fragmentation of cyclic protein. Protein Sci. 7, 1583 1592. Tam, J.P., Yu, Q., 1998. Methionine ligation strategy in the biomimetic synthesis of parathyroid hormones. Biopolymers 46, 319 327. Tam, J.P., Zavala, F., 1989. Multiple antigen peptide. A novel approach to increase detection sensitivity of synthetic peptides in solid-phase immunoassays. J. Immunol. Methods 124, 53 61. Tam, J.P., Clavijo, P., Lu, Y.-A., Nussenzweig, V., Nussenzweig, R., Zavala, F., 1990. Incorporation of T and B epitopes of the circumsporozoite protein in a chemically dened synthetic vaccine against malaria. J. Exp. Med. 171, 299 306.
228
K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229 Vanslyke, J.K., Hruby, D.E., 1994. Immunolocalization of vaccinia virus structural proteins during virion formation. Virology 198, 624 635. Vilaseca, L.A., Rose, K., Werlen, R. et al., 1993. Protein conjugates of dened structure: synthesis and use of a new carrier molecule. Bioconj. Chem. 4, 515 520. Vogel, T., Kurth, R., Norley, S., 1994. The majority of neutralizing Abs in HIV-1-infected patients recognize linear V3 loop sequences. Studies using HIV-1MN multiple antigenic peptides. J. Immunol. 153, 1895 1904. Wallace, A., Altamura, S., Toniatti, C. et al., 1994. A multimeric synthetic peptide combinatorial library. Pept. Res. 7, 27 31. Wang, C.Y., Looney, D.J., Li, M.L. et al., 1991. Long-term high-titer neutralizing activity induced by octameric synthetic HIV-1 antigen. Science 254, 285 288. Wang, R., Charoenvit, Y., Corradin, G. et al., 1995. Induction of protective polyclonal antibodies by immunization with a Plasmodium yoelii circumsporozoite protein multiple antigen peptide vaccine. J. Immunol. 154, 2784 2793. Weeks, B.S., Nomizu, M., Otaka, A. et al., 1994. Lymphocytes and promonocytes attach to the synthetic wTyr5,12,Lys7xpolyphemusin II peptide. Biochem. Biophys. Res. Commun. 202, 470 475. Whittaker, R.G., Hayes, P.J., Bender, V.J., 1993. A gentle method for linking Tris to amino acids and peptides. Pept. Res. 6, 125 128. Widmann, C., Romero, P., Maryanski, J.L., Corradin, G., Valmori, D., 1992. T helper epitopes enhance the cytotoxic response of mice immunized with MHC class I-restricted malaria peptides. J. Immunol. Methods 155, 95 99. Wilson, B.S., Nuoffer, C., Meinkoth, J.L. et al., 1994. A Rab1 mutant affecting guanine nucleotide exchange promotes disassembly of the Golgi apparatus. J. Cell Biol. 125, 557 571. Wo, Z.G., Oswald, R.E., 1995. A topological analysis of goldsh kainate receptors predicts three transmembrane segments. J. Biol Chem. 270, 2000 2009. Wong, F., Schaefer, E.L., Roop, B.C., LaMendola, J.N., Johnson-Seaton, D., Shao, D., 1989. Proper function of the Drosophila trp gene product during pupal development is important for normal visual transduction in the adult. Neuron 3, 81 94. Wrighton, N.C., Balasubramanian, P., Barbone, F.P. et al., 1997. Increased potency of an erythropoietin peptide mimetic through covalent dimerization. Nat. Biotechnol. 15, 1261 1265. Wunsch, E., Morodor, L., Nyfeler, R., Kalbacher, H., Gemeimer, M., 1985. Immunodetermination of peptide factors. I. Synthesis of N-alpha-maleoyl-peptide derivatives. Biol. Chem. Hoppe-Seyler 366, 53 61. Yahi, N., Sabatier, J.M., Nickel, P., Mabrouk, K., GonzalezScarano, F., Fantini, J., 1994a. Suramin inhibits binding of the V3 region of HIV-1 envelope glycoprotein gp120 to galactosylceramide, the receptor for HIV-1 gp120 on human colon epithelial cells. J. Biol. Chem. 269, 24349 24353.
Tam, J.P., Lu, Y.-A., Liu, C.-F., Shao, J., 1995. Peptide synthesis using unprotected peptides through orthogonal coupling methods. Proc. Natl. Acad. Sci. USA 92, 12485 12489. Tam, J.P., Lu, Y.-A., Yu, Q., 1999. Thia zip reaction for synthesis of large cyclic peptides: mechanisms and applications. J. Am. Chem. Soc. 121, 4316 4324. Tam, J.P., Yu, Q., Miao, Z., 2000. Orthogonal ligation strategies for peptide and protein. Biopolymers 51, 311 332. Tam, J.P., Xu, J., Eom, K.D., 2001. Methods and strategies of peptide ligation. Biopolymers, in press. Thakur, S., Vadolas, D., Germann, H.P., Heidemann, E., 1986. Inuence of different tripeptides on the stability of the collagen triple helix. II. An experimental approach with appropriate variations of a trimer model oligotripeptide. Biopolymers 25, 1081 1086. Toth, G.K., Varadi, G., Nagy, Z. et al., 1993. Branched polypeptides as antigens for inuenza virus hemagglutinin and T-cell receptor subunits. Pept. Res. 6, 272 280. Toth, I., Sakthivel, T., Wilderspin, A.F. et al., 1999. Novel cationic lipidic peptide dendrimer vectors In vitro gene delivery. S.T.P. Pharma. Sci. 9, 93 99. Tres, L.L., Kierszenbaum, A.L., 1996. Sak57, an acidic keratin initially present in the spermatid manchette before becoming a component of paraaxonemal structures of the developing tail. Mol. Reprod. Dev. 44, 395 407. Troalen, F., Razandratsita, A., Puisieux, A. et al., 1990. Structural probing of human lutropin using antibodies raised against synthetic peptides constructed by classical and multiple antigen peptide system approaches. Mol. Immunol. 27, 363 368. Tsikaris, V., Sakarellos, C., Cung, M.T., Marraud, M., Sakarellos-Daitsiotis, M., 1996. Concept and design of a new class of sequential oligopeptide carriers SOC. for covalent attachment of multiple antigenic peptides. Biopolymers 38, 291 293. Unson, C.G., Erickson, B.W., Richardson, D.G., Richardson, J.S., 1984. Protein engineering: design and synthesis of a protein. Fed. Proc. 43, 1837. Valmori, D., Pessi, A., Bianchi, E., Corradin, G., 1992. Use of human universally antigenic tetanus toxin T cell epitopes as carriers for human vaccination. J. Immunol. 149, 717 721. Valmori, D., Romero, J.F., Men, Y., Maryanski, J.L., Romero, P., Corradin, G., 1994. Induction of a cytotoxic T cell response by co-injection of a T helper peptide and a cytotoxic T lymphocyte peptide in incomplete Freunds adjuvant IFA.: further enhancement by pre-injection of IFA alone. Eur. J. Immunol. 24, 1458 1462. Vanage, G., Lu, Y.-A., Tam, J.P., Koide, S.S., 1992. Infertility induced in rats by immunization with synthetic peptide segments of a sperm protein. Biochem. Biophys. Res. Commun. 183, 538 543. Vanage, G.R., Jaiswal, Y.K., Lu, Y.-A., Tam, J.P., Wang, L.F., Koide, S.S., 1994. Immunization with synthetic peptide segments of a sperm protein impair fertility in rats. Res. Commun. Chem. Pathol. Pharmacol. 84, 3 15.
K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229 Yahi, N., Fantini, J., Mabrouk, K. et al., 1994b. Multibranched V3 peptides inhibit human immunodeciency virus infection in human lymphocytes and macrophages. J. Virol. 68, 5714 5720. Yahi, N., Sabatier, J.M., Baghdiguian, S., Gonzalez-Scarano, F., Fantini, J., 1995. Synthetic multimeric peptides derived from the principal neutralization domain V3 loop. of human immunodeciency virus type 1 HIV-1. gp120 bind to galactosylceramide and block HIV-1 infection in a human CD4-negative mucosal epithelial cell line. J. Virol. 69, 320 325. Yajima, H., Fujii, N., 1981. Studies on peptides. 103. Chemical synthesis of a crystalline protein with the full enzymatic-activity of ribonuclease-a. J. Am. Chem. Soc. 103, 5867 5871. Yamashiro, D., Li, C.H., 1988. New segment synthesis of alpha-inhibin-92 by the acyl disulde method. Int. J. Pept. Protein Res. 31, 322 334. Yang, Y.Y., Johnson, T.M., Mecham, J.O., Tam, J.P., Li, J.K., 1992. Epitopic mapping of linear and conformation-dependent antigenic determinants on GP5 of ve US bluetongue viruses. Virology 188, 530 536. Yang, H.W., Ikeda, H., Kato, K. et al., 1999. Development of a polyclonal antibody with dened specicity against synthetic peptides from the N-myc oncoprotein using multiple antigen peptide and hemocyanin conjugation methods. J. Pediatr. Surg. 34, 454 460. Yao, Z.J., Kao, M.C., Loh, K.C., Chung, M.C., 1994. Antibody against branched epitope as an afnity ligand to separate the parent protein. J. Chromatogr. A 679, 190 194. Yoshitake, S., Yamada, Y., Ishikawa, E., Masseyeff, R., 1979. Conjugation of glucose oxidase from Aspergillus niger and rabbit antibodies using N-hydroxysuccinimide ester of N4-carboxycyclohexylmethyl.-maleimide. Eur. J. Biochem. 101, 395 399. Zanini, D., Roy, R., 1998. Practical synthesis of starburst PAMAM-thiosialodendrimers for probing multivalent carbohydrate lectin binding properties. J. Org. Chem. 63, 3486 3491. Zavala, F., Chai, S., 1990. Protective anti-sporozoite antibodies induced by a chemically dened synthetic vaccine. Immunol. Lett. 25, 271 274.
229
Zelezna, B., Richards, E.M., Tang, W., Lu, D., Sumners, C., Raizada, M.K., 1992. Characterization of a polyclonal antipeptide antibody to the angiotensin II type-1 AT1. receptor. Biochem. Biophys. Res. Commun. 183, 781 788. Zelezna, B., Veselsky, L., Velek, J., Zicha, J., Kunes, J., 1994. Angiotensin AT1 receptor blockade by specic antibody prevented two-kidney, one-clip renal hypertension in the rat. Eur. J. Pharmacol. 260, 95 98. Zeng, W., Jackson, D.C., Murray, J., Rose, K., Brown, L.E., 2000. Totally synthetic lipid-containing polyoxime peptide constructs are potent immunogens. Vaccine 18, 1031 1039. Zhang, L., Tam, J.P., 1996. Thiazolidine formation as a general and site-specic conjugation method for synthetic peptides and proteins. Anal. Biochem. 233, 87 93. Zhang, L., Tam, J.P., 1997a. Orthogonal coupling of unprotected peptide segments through histidyl amino terminus. Tetrahedon Lett. 38, 3 6. Zhang, L., Tam, J.P., 1997b. Synthesis and application of unprotected cyclic peptides as building blocks for peptide dendrimers. J. Am. Chem. Soc. 119, 2363 2370. Zhang, L., Torgerson, T., Liu, X.-Y. et al., 1998. Preparation of functionally active cell-permeable peptides by single-step ligation of two peptide modules. Proc. Natl. Acad. Sci. USA 95, 9184. Zhong, G., Toth, I., Reid, R., Brunham, R.C., 1993. Immunogenicity evaluation of a lipidic amino acid-based synthetic peptide vaccine for Chlamydia trachomatis. J. Immunol. 151, 3728 3736. Zhou, F.C., Xu, Y., Bledsoe, S., Lin, R., Kelley, M.R., 1996. Serotonin transporter antibodies: production, characterization, and localization in the brain. Brain Res. Mol. Brain Res. 43, 267 278. Zhou, S.R., Whitaker, J.N., 1996. Active immunization with complementary peptide PBM 9-1: preliminary evidence that it modulates experimental allergic encephalomyelitis in PLrJ mice and Lewis rats. J. Neurosci Res. 45, 439 446. Zolnierowicz, S., Csortos, C., Bondor, J., Verin, A., Mumby, M.C., DePaoli-Roach, A.A., 1994. Diversity in the regulatory B-subunits of protein phosphatase 2A: identication of a novel isoform highly expressed in brain. Biochemistry 33, 11858 11867.

Peptide Dendrimers Aplication and Synthesis

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Reviews in Molecular Biotechnology 90 2002.

Peptide dendrimers: applications and synthesis

K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229

K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229

Fig. 1. Schematic illustration of peptide dendrimers that vary in size.

K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229

Fields et al. 1993. Hahn et al. 1990.

K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229

enhanced and a potentially useful therapeutic agent produced.

K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229

K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229

CS protein, P. malariae CS protein, P. falciparum

204 Table 3 Continued. Target CS protein, P. yoelii

K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229

K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229

K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229

K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229

Fig. 3. A schematic comparison of divergent and convergent synthesis methods.

K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229

Fig. 4. Examples of simple amino, carboxylic and hydroxylic cores.

K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229

K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229

K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229

Fig. 7. Solid phase synthesis of a K 2 K glyoxylyl core.

K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229

Fig. 8. Synthesis of thiol-functionalized S-acetyl and CysNpys. di-K 2 K cores.

K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229

K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229

Fig. 11. Convergent synthesis of an articial hemeprotein.

K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229

K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229

K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229

K. Sadler, J.P. Tam r Re iews in Molecular Biotechnology 90 (2002) 195 229

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