22

Multiplex Immunoassay
and Bead Based Multiplex
Türkan Yiğitbaşı
Izmir Katip Çelebi University
Turkey

1. Introduction
1.1 Background
Protein immunoassays provide information about quantities and forms of endogenous
proteins. Uniplex enzyme immunoassays like Elisa have been the workhorse for protein
measurement for decades, but they can be laborious and expensive and consume relatively
large amounts of specimen. In comparison to the ELISA for a single analyte, multiplex
assays offer the possibility of obtaining more reliable quantitative information in a highly
parallel analysis. In addition, quantitative multiplexed assays offer the possibility to identify
combinations of biomarkers with higher disease specificity than any single established
biomarker alone. Because of these reasons, currently one-analysis, many-metabolites, many-
diseases approach is receiving more attention than uniplex system with one-analysis, one-
metabolite, one-disease.

1.2 Multiplex immunoassay platforms

Presently, antibody-based platforms are the core technology for protein multiplex arrays.
Assay formats include suspension arrays (microbead assays) and planar arrays that use
traditional immunometric principles.
Planar arrays: MULTI-ARRAY (Meso Scale Discovery), A2 (Beckman Coulter), and FAST
Quant (Whatman Schleicher & Schuell BioScience), Searchlight (Aushon Biosystem).
Suspension arrays: Bio-Plex (Bio-Rad Laboratories), FlowCytomix (Bender MedSystems),
cytometric bead array (Becton, Dickinson and Company) and the partners of Luminex Corp
(XMAP)].
In the first format, different capture antibodies are spotted at defined positions on a 2-
dimensional array. In the second, the capture antibodies are conjugated to different
populations of microbeads, which can be distinguished by their fluorescence intensity in a
flow cytometer.
Optimal assay performance of multiplex immunoassay platforms depend on proprietary
information about the antibody pair, the composition of diluents, and the software. The
accuracy of quantification for multiplexed immunoassays depends, as with all ELISAs, on
the quality of the calibration curves, assay imprecision (CV), recoveries, and assay linearity

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352 Trends in Immunolabelled and Related Techniques

(the limits of quantification). Therefore, there is not a single assay platform suitable for all
analytes. The selection of a platform or kit should depend on assay sensitivity, the relevant
biological concentration to be measured

Platform MesoScale discovery Searchlight FastQuant

(MULTI- (FAST Quant System)
ARRAY/MULTI-SPOT)
Capture antibody Carbon Plastic Nitrocellulose
binding surface
Detection system Electrochemiluminescent Biotinylated Biotinylated detector
detector with with fluorescent
fluorescent detection
detection
Analytes/plex Up to 10 Up to 24 Up to 10
Image System CCD camera based CCD camera CCD camera based
based
Customised array Yes Yes Yes
Kits for designing Yes No No
and building
your own assay
Reagent company Waterman Scleicher& Aushon Waterman Scleicher&
Schuell Bioscience Biosystem Schuell Bioscience
Commercial Waterman Scleicher& Aushon Waterman Scleicher&
instrument Schuell Bioscience Biosystem Schuell Bioscience
Table 1. The characteristics of some commercial planar microspot array platforms

Platform Luminex Cytometric Bead Array (CBA)

Capture antibody Fluorescently tagged beads Fluorescently labeled beads
binding surface
Detection system Biotinylated detector with Phycoerythrin conjugated
fluorescent detection detectors
Analytes/plex Up to 100 Up to 100
Image System Luminex XMAP based system Flow cytometer with dual laser
Customised array Yes Yes
Kits for designing Yes Yes
and building your
own assay
Reagent company Luminex Corp and its BD Becton,Dickinson and others
partners(many others)
Commercial Lumine(XMAP) and its partners BD Becton,Dickinson and others
instrument
Table 2. The characteristics of some commercial suspension bead array platforms

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Multiplex Immunoassay and Bead Based Multiplex 353

Fig. 1. Process of multiplexed immunodetection using fluorescent-coded beads

(www.millipore.com)

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354 Trends in Immunolabelled and Related Techniques

2. Bead based multiplex

2.1 Background
In multiplex bead array assays (MBAA), beads of discrete fluorescence intensities and
wavelengths provide a capture surface for specific analytes enabling detection of multiple
analytes in a single sample. Bead based assays of analyte is an attractive strategy for
obtaining large numbers of measurements rapidly. MBAA is an open access system; namely,
the parameters to be measured can be determined by the user. The variety of source
increases its area of application (nucleic acids, antigens, antibodies, receptors).Since the
detection method is based on flow cytometry and this allows repeatability, MBAA performs
the quality control of the measurement, simultaneously with the measurement itself.
The MBAA readers allow analysis of up to 100 reactions or assays per well. This remarkable
capability allows incorporating a large number of assays, such as DNA, receptor-ligand,
Immunoassay and enzyme in life science research, drug discovery as well as diagnostic areas.

2.2 The principle of measurement

These systems depend on measurement of fractional antibody occupancy using two different
labels: one labeling the “capture” antibody, the second labeling a “detection antibody”,
selected to react either with occupied or unoccupied sites on the “capture” antibody. The ratio
of signals emitted by the two labeled antibodies reveals the analyte concentration to which the
capture antibody has been exposed. An array of capture antibodies, each labeled with the
same fluorescent label, is scanned (by a laser), and the fluorescent signal ratio emitted from
each discrete antibody couplet in the array is measured.

2.3 Summary of principles

1. Color-coded beads, pre-coated with analyte-specific capture antibody for the molecule
of interest are added.
2. Analyte-specific antibodies capture the analyte of interest. Biotinylated detection
antibodies specific to the analyte of interest are added and form an antibody-antigen
sandwich.
3. Phycoerythrin (PE)-conjugated Streptavidin is added.
4. The beads are read by a dual-laser flow-based detection instrument which excites the
internal dyes marking the beads set and a second laser excites PE, the fluorescent dye
on the reporter molecule (www.luminexcorp.com).
The system is capable of measuring potentially up to 100 analytes simultaneously in a small
sample volume (25–50 µL).

2.4 Advantages of the system

1. Speed/High Throughput — Because each microsphere serves as an individual test, a
large number of different assays can be performed and analyzed simultaneously
2. Versatility — Bead based multiplex system can perform assays in several different
formats, including nucleic acids and antigen-antibody binding, along with enzyme,
receptor-ligand and other protein interactions

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Multiplex Immunoassay and Bead Based Multiplex 355

3. Flexibility — The technology can be customized for the user’s specific needs based
upon their analytes of interest
4. Accuracy — The technology generates real-time analysis and accurate quantification of
the biological interactions (www.luminexcorp.com).
Advantages of this approach also include specimen conservation, limited sample handling,
and decreased time and cost. However, it is difficult to optimize assay format for each protein,
to select common dilution factors and to establish reliable quality control algorithms.

Characteristics Antibody-based
Principle Antibody–antigen interaction
Required reagents/information Specific antibody pairs
Quantification basis flowcytometer and others
Multiplexicity 1–100
Clinical diagnostic test Yes
Sample enrichment No
Sensitivity >sub ng/mL–pg/mL
Reproducibility Excellent
Assay development Time and resource demanding
Consumable High demand
Robustness Yes
Throughput High
Matrix effect Yes
Sample manipulation No
Automation Yes
History >15 years
Table 3.The characteristics of MBAA

2.5 Comparison with other technologies and assays

The assay which is most often compared to MBAA is the enzyme-linked immunosorbent
assay (ELISA). ELISAs, in general, use a similar immobilized antibody to capture a soluble
ligand, with subsequent detection of the captured ligand by a second antibody. There are,
however, several substantial differences between MBAA and ELISAs. For example, MBAA
uses fluorescence as a detection system where ELISAs use enzyme amplification of a
colorimetric substrate. MBAA captures ligands onto spherical beads in suspension while
ELISAs generally rely upon flat surfaces in 96-well plates. Most importantly, MBAA
techniques, by their very nature, are multiplexed and therefore may be subject to any
perturbations that arise from analyzing multiple ligands simultaneously, such as cross-
reactivities. By contrast, ELISA methodologies generally study one analyte at a time, and
thus avoid any concerns arising from multiplexing.
Multiplex system presents additional Quality control (QC) challenges compared to uniplex
analyses. The failure of 1 constituent assay to meet QC specifications results in rejection of
results for all assays on the panel. Samples failing QC specifications should be retested using
the same measurement system, because substitution of a uniplex assay may introduce bias

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356 Trends in Immunolabelled and Related Techniques

due to differences in assay format. However, the probability of all assays simultaneously
meeting QC specifications is much lower than the probability of a uniplex test passing QC.
At present, reference guidelines for multiplex QC programs are under development by the
Food and Drug Administration (FDA).
In theory, the MBAA platform may also provide a wider dynamic range than conventional
ELISA methods because of the greater linear range of fluorescence intensity compared with
absorbance. Detection limits of MBAA (luminex) and ELISA are different. A comparison of
detection limits of some test for the Luminex assays and the ELISAs is shown in Table 4. Data
Supplement (available at http://www.clinchem.org/content/vol51/issue7/) Disadvantage of
MBAA are time consuming in assay development, lacking antibody pairs for new biomarkers
and limited commercial multiplex assay kits.

Multiplex panels Analytes Luminex ELISA ELISA

manufacturer
Linco Endocrine Leptin 0.13 µg/L 1.39 µg/L Linco
Insulin 0.31 mIU/L 0.14 mIU/L Alpco
C-peptide 0.037 µg/L 0.132 µg/L Linco
Biosource Chemokine MCP-1 2.35 ng/L 2.57 ng/L R&D
Eotaxin 4.37 ng/L 3.80 ng/L R&D
Upstate Chemokine MCP-1 20.5 ng/L 2.57 ng/L R&D
Eotaxin 27.72 Eotaxin 3.80 ng/L R&D
Linco Cytokine TNF-α 1.61 ng/L 0.092 ng/L R&D
IL-8 2.52 ng/L 0.26 ng/L Biosource
Upstate Cytokine TNF-α 0.065 ng/L 0.092 ng/L R&D
IL-8 0.70 ng/L 0.26 ng/L Biosource
IL-6 0.75 ng/L 0.068 ng/L R&D
R&D Cytokine TNF-α 1.53 ng/L 0.092 ng/L R&D
IL-6 0.977 ng/L 0.068 ng/L R&D
Table 4. Comparison of detection limits for the Luminex multiplexed assays and the
individual ELISAs.

Protein microarray kits, which use capture antibodies and detection antibodies in a
multiplex fashion similar to MBAA, should also be considered as a competing technology
which is most commonly used for simultaneous determination of multiple proteins in a
biological fluid. The technique uses primary antibodies as the immobilized probe on a solid
surface, and protein antigens labeled with fluorophores with or without bound secondary
antibodies are recognized and detected. However, binding of antibodies and antigens to a
solid support can cause denaturation or drying of proteins. MBAA provides multiplexing in
a solution phase and thus is particularly flexible and nondestructive for protein analysis.
Also protein microarray assay are relatively new, are not widely accepted as a ‘gold
standard’ for clinical use, and also may be of limited sensitivity.

2.6 Areas of application

Since a disease can result from various reasons and it may include functional disorders of
multiple genes, a multi analyte analysis is necessary for diagnostic purposes. The

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Multiplex Immunoassay and Bead Based Multiplex 357

technology called omic and multiplex system allows a fast and systematic detection of the
effects of micro molecules in different molecular and cellular contexts. In this respect, it can
be used for diagnosing multi-reason/multi-gene diseases, performing a comprehensive
disease management and investigating complex cellular functions. In addition, it provides a
wide range of application area for research area because of its plasticity which allows the
researchers to perform various studies.

2.7 Clinical utility

Despite the introduction of hundreds of multiplexed protein immunoassays to the research
market in recent years, only a limited number have been cleared by the FDA for clinical use,
an observation that illustrates the complexity of constructing robust arrays. Antibody-based
multiplexed assay (and commercial instruments and kits) have a relatively long history with
over 15 years of development and optimization. However, most commercial multiplex
assays are developed for research laboratories and nonclinical tests; only a limited number
of multiplex assays are approved by FDA for clinical testing. For example FDA-cleared
planar protein multiplex arrays consist primarily of the lateral flow immunoassays used for
point-ofcare evaluation . For example, the Triage® Cardio ProfilER® 4-plex measures
troponin-I, creatine kinase-MB, myoglobin, and brain natriuretic peptide (BNP) to assist
with evaluation of chest pain using a portable lateral flow platform. At present, suspension
immunoassay is the prevailing technology for FDA-cleared multiplex protein
measurements, especially for testing antibodies in the serum of patients with allergies or
autoimmune or infectious diseases in clinical laboratories.
The other platforms for commercial multiplex diagnostic tests include Luminex
(www.luminex.com), Triage system (www.Biosite.com), Evidence (www.Randox.com),
Vidas (www.biomerieux-diagnostics. com), Planner arrays (www.VBC-genomics),Whatman
(www.whatman.com) and bead array (www.bioarrays.com). At present, suspension
immunoassay is the prevailing technology for FDA-cleared multiplex protein
measurements, especially for testing antibodies in the serum of patients with allergies or
autoimmune or infectious diseases in clinical laboratories. Most of the currently available
multiplex immunoassays have been designed to quantify the concentrations of various
cytokines.The recent development of spectrally-distinguishable fluorescent beads(Luminex)
(Kellar and Iannone, 2002) has resulted in the widespread use of antigen-coupled beads for
monitoring antibodies in sera by flow cytometry. These bead arrays have been adapted for
serologic screening of antigens and have been described for up to ten antigens for HIV (Opalka
et al., 2004), HPV (Opalka et al., 2003; Dias et al., 2005), Epstein-Barr virus (Klutts et al., 2004;
Binnicker et al., 2008; Gu et al., 2008), B. anthracis (Biagini et al., 2004; Biagini et al., 2005),
Influenza (Drummond et al., 2008) and M.tuberculosis (Khan et al.,2008).
The results from a growing number of research studies, demonstrate that multiplex
technology may be useful in clinical research to measure a large number of analytes to
examine the association with a clinical phenotype and the effects of therapeutic
interventions, and that this technology may be particularly useful when sample volume is
limited, such as in large epidemiologic studies and clinical trials. Clinical applications must
follow establishment of globally accepted calibration standards, performance criteria, and
QC programs.

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358 Trends in Immunolabelled and Related Techniques

APPLICATION AVAILABLE KITS* COMPANY

Allergy Testing Alternaria (Mold) (h), Bermuda Grass (h), Cat Dander (h), Egg ImTech (h)
White (h), Milk (h), Mite Pternoyssinus (h), Mountain Cedar (h),
Short Ragweed (h), Timothy Grass (h), Wheat (food) (h)
Autoimmune ASCA (h), beta-2 Microglobulin (h,m), Centromere B (h), RBM(h,m)
Chromatin (h), DNA (h),ENA Profile 4 (SSA, SSB, Sm, ENA Profile
5 SSA, SSB, Sm, RNP, Scl-70) (h), ENA Profile 6 (SSA, SSB, Sm,
RNP, Scl-70, Jo-1) (h), Gliadin A (h), Gliadin G (h), Histone (h),
Histone H1 (h), Histone H2A (h), Histone H2B (h), Histone H3 (h),
Histone H4 (h), HSP-27 pS82 (G), HSP-27 Total (G), HSP-32 (h),
HSP-65 (h), HSP-71 (h), HSP-90 a (h), HSP-90 b (h), Jo-1 (h), PCNA
(h,m), PR3 (h), PR3 (cANCA) (m), RF (h), Ribosomal P (h,m), RNP
(h,m), RNP-A (h), RNP-C (h), SCF (h,m), Scl-70 (h,m), Serum
Amyloid P (h), SLE Profile 8 (SSA, SSB, Sm, RNP, Scl-70, Jo-1,
Ribosome-P, chromatin) (h), Sm (G) (h), Smith (h,m), SSA (h,m),
SSB (h,m), Streptolysin O (h), TG (h),
TPO (h,m), Transglutaminase A (h), Transglutaminase G (h)
Cancer Markers Alpha Fetoprotein (h), Cancer Antigen 125 (h), Carcinoembryonic RBM(h)
Antigen (h), PSA, Free (h)
Cardiac Markers Creatine Kinase-MB (h), Endothelin-1 (m), PAP (h), SGOT (h,m), RBM(h,m)
TIMP-1 (h,m)
Cytokine Abeta 40 (h), Abeta 42 (h), BDNF (h), DR-5 (h), EGF (h,m), ENA-78 B-R(m);
(h), Eotaxin (h,m), Fatty Acid Binding Protein (h), FGF-basic (h,m), Bios (h,m,rt);
G-CSF (h,m), GCP-2 (m), GM-CSF (h,m,rt), GRO alpha (h), GRO- Linco ( h,m,rt);
KC (rt), HGF (h,m), I-TAC (h), ICAM-1 (h), IFN-alpha (h), IFN- RD(h,m);
gamma (h,m,rt), IL-10 (h,m,rt), IL-11 (m), IL-12 (h,m), IL-12 p40 UP(h,m), RBM
(h,m), IL-12 p40/p70 (m) (rt), IL-12 p70 (h,m,rt), IL-13 (h,m), IL-15 (h,m)
(h,m), IL-16 (h), IL-17 (h,m), IL-18 (rt), IL-1alpha (h,m,rt), IL-1beta
(h,m,rt), IL-1ra (h), IL-1ra/IL-1F3 (h), IL-2 (h,m,rt), IL-3 (h,m), IL-4
(h,m,rt), IL-5 (h,m,rt), IL-6 (h,m,rt), IL-7 (h,m), IL-8 (h), IL-9 (m), IP-
10 (h,m), JE/MCP-1 (m), KC (m), KC/GROa (m), LIF (m), IL-8 (h),
IL-9 (m), IP-10 (h,m), JE/MCP-1 (m), KC (m), KC/GROa (m), LIF
(m), MCP-3(h,m), MCP-5 (m)
EndocrineACTH ACTH (h), Adiponectin (h,m), Amylin (m) (rt) (h), C-Peptide (h), Linco (h,m,rt);
(h), Adiponectin Calcitonin (h), CRF(h), FGF-9 (m), FSH (h), GH (h), GLP-1 (h,m,rt), RBM ( h,m)
(h,m), Amylin Glucagon (m) (rt) (h), Growth Hormone (h,m), Insulin (h,m,rt),
(m) (rt) (h), C- Leptin (h,m,rt), LH (h), Lipoprotein (a) (h), PAI-1(active) (h), PAI-1
Peptide (h), (total) (h,m), Prolactin (h), Resistin (h,m,rt), T3 (h), T4 (h), TBG (h),
Calcitonin (h), Thyroglobulin (h), TSH (h)
CRF Linco
Gene Expression IL6R(h), ACTB (h), BAD (h), BAK1 (BAK) (h), BCL2 (h), BCL2L1 Bios (h);
(BCL-XL) (h), CDKN1A (CDKN1) (h), CFLAR (CFLIP) (h), CSF2 MBio (h)
(h), GAPD (h), IFN-gamma (h), IL-1 beta (h), IL-10 (h), IL-2 (h), IL-6
(h), IL-8 (h), NFKB2 (h), NFKBIA (NFKIA) (h), NKFB1 (h), PPIB
(h), Ptk2B (RAFTK) (h), RELA (h), RELB (h), TNF (h), TNFAIP3
(A20) (h), TNFRSF6 (FAS) (h), TNFSF6 (FASL) (h), VEGF (h)
MMP MMP-1 (h), MMP-12 (h), MMP-13 (h), MMP-2 (h), MMP-3 (h), RD (h); Bios (h)
MMP-7 (h), MMP-8(h), MMP-9 (h)
Genotyping FlexMAP™ (G), Mitochondrial DNA Screening (h), Tag-It™ Bio (h), Mira(h),
Mutation Detection Kit(G), Y-SNP Identification (h) TmBio (h)

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Multiplex Immunoassay and Bead Based Multiplex 359

APPLICATION AVAILABLE KITS* COMPANY

Infectious Adenovirus (h,m), Bordetella pertussis (h), Campylobacter jejuni RBM (h,m)
Disease (h), Chlamydia pneumoniae (h), Chlamydia trachomatis (h),
Cholera Toxin (h), Cholera Toxin b (h), Clostridium piliforme
(Tyzzer's) (m), Cytomegalovirus (h,m), Diphtheria Toxin (h),
Ectromelia virus (m), EDIM (Epidemic diarrhea of infant mice) (m),
Encephalitozoon cuniculi (m), Epstein-Barr EA (h), Epstein-Barr
NA (h), Epstein-Barr VCA (h), HBV Core (h), HBV Envelope (h),
HBV Surface (Ad) (h), HBV Surface (Ay) (h), HCV Core (h), HCV
NS3 (h), HCV NS4 (h), HCV NS5 (h), Helicobacter pylori (h),
Hepatitis A (h), Hepatitis D (h), HEV orf2 3KD (h), HEV orf2 6KD
(h), HEV orf3 3KD (h), HIV-1 gp120 (h), HIV-1 gp41 (h), HIV-1 p24
(h), HPV (h), HSV-1 gD (h), HSV-1/2 (h), HSV-2 gG (h), HTLV-1/2
(h), Influenza A (h), Influenza A H3N2 (h), Influenza B (h),
Leishmania donovani (h), Lyme disease (h), Lymphocytic
choriomeningitis virus (m), M. pneumoniae (h), M. tuberculosis (h),
Minute virus (m), Mumps (h), Mycoplasma pulmonis (m),
Parainfluenza 1 (h), Parainfluenza 2 (h), Parainfluenza 3 (h),
Parvovirus (m), Pneumonia virus of mice (m)
Isotyping IgA (h,m), IgE (h,m), IgG1 (m), IgG2alpha (m), IgG2beta (m), IgG3 UP (h,m);
(m), IgM (h,m), light chain (kappa or gamma) (m) RBM(h,m)
Metabolic Apolipoprotein A-1 (m), Apolipoprotein A-I (h), Apolipoprotein RBM (h,m),
Markers A-II (h), Apolipoprotein B (h), Apolipoprotein C-II (h), Linco
Apolipoprotein C-III (h), Apolipoprotein E (h), beta-2 Glycoprotein
(h,m), Collagen Type 1 (h), Collagen Type 2(h), Collagen Type 4 (h),
Collagen Type 6 (h), Glutathione S-Transferase (h,m),
Pancreatic Islet Cells (h), tTG (Celiac Disease) (h)
Tissue Typing HLA Class I and II (h), HLA Class I Single Antigen Antibody, Lambda (h)
Group 1 (h), HLA Class I Single Antigen Antibody, Group 2 (h),
PRA Class I (h), PRA Class I and II (h), PRA Class II (h), SSO Class I
HLA-A (h), SSO Class I HLA-B (h), SSO Class I HLA-C(h), SSO
Class II DP (h), SSO Class II DQB1 (h), SSO Class II DRB1 (h), SSO
Class II DRB3,4,5 (h)
Kinase Akt (G), Akt (Ser473) (G), Akt (total) (G), Akt/PKB (total) (G), UP (G), Bios (G)
Phosphorylated Akt/PKBpS473 (G), ATF2 (Thr71) (G), ATF2 (total) (G), CREB
Protein (pS133) (G), CREB (Total) (G), Erk 1/2(pTpY185/187) (G), Erk 1/2
(Total) (G), Erk-2 (G), Erk1 (Thr202/Tyr204) (G), Erk1/2
(Thr202/Tyr204, Thr185/Tyr187) (G), Erk2 (Thr185/Tyr187) (G),
Erk2 (total) (G),GSK 3beta (pS9) (G), GSK-3a/b (Ser21/Ser9) (G),
GSK-3beta (G), IGF 1R
Transcription AP-2 (G), CREB (G), EGR (h), HIF-1 (h), NF-1 (h), NFAT (h), NFkB Bios (h), MBio
Factors-Nuclear Gene Family(h), PPAR (h), SRE (h), YY1 (h) (h)
Receptors

* Human (h), mouse (m): rat (rt): general (G), ImmuneTech (ImTech); Rules Based Medicine (RBM); Bio-
Rad (B-R); BioSource (Bios); Linco Research (Linco); Qiagen; R&D Systems (RD); and Upstate Group
(UP); Marligen Biosciences (MBio), MiraBio (Mira); Tm BioScience (TmBio); One Lambda (LAMBDA).

Table 5. Aplication area and available kits

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360 Trends in Immunolabelled and Related Techniques

3. References
[1] Multiplexed Analysis of Biomarkers Related to Obesity and the Metabolic Syndrome in
Human Plasma, Using the Luminex-100 System. Liu M.Y, Xydakis A.M,
Hoogeveen R.C, Jones P.H, O’Brian Smith E, Nelson K.W, Ballantyn C.M. Clinical
Chemistry.2005;51(7):1102–1109 .
[2] Multiplex assays for biomarker research and clinical application: Translational science
coming of age. Fu Q, Schoenhoff F.S, Savage W.J, Zhang P, Van Eyk J.E.Proteomics
Clin. Appl. 2010;4:271–284.
[3] Comparison of multiplex immunoassay platforms. Fu Q, Zhu J, Van Eyk JE. Clin Chem.
2010;56(2):314-8.
[4] Measurement and Quality Control Issues in Multiplex Protein Assays: A Case Study.
Ellington A.A, Kullo I.J, Bailey K.R, Klee G.G. Clin Chem. 2009; 55(6): 1092–1099.
[5] Multi-analyte immunoassay. Ekins RP.Journal of Pharmaceutical and Biomedical
Analysis.1989; 7(2):155-168.
[6] US Food and Drug Administration. Guidance for industry and FDA staff:
pharmacogenetic tests and genetic tests for heritable markers. (Accessed October
2011].
[7] Multiplexed protein measurement: technologies and applications of protein and
antibody arrays. Kingsmore S.F. Nat Rev Drug Discov. 2006; 5(4): 310–3208.
[8] Antibody-Based Protein Multiplex Platforms: Technical and Operational Challenges.
Ellington A.A, Kullo IJ, Bailey K.R, Klee G.G. Clin Chem. 2010; 56(2): 186–193.
[10] Rapıd Detectıon of Antıbodıes In Sera Usıng Multıplexed Self-Assemblıng Bead Arrays.
Wonga J, Sibanib S, Lokkoa N.N, LaBaerb J, Andersona K. J Immunol Methods.
2009; 31: 171–182.
[11] Multiplex Bead Array Assays: Performance Evaluation and Comparison of Sensitivity
to ELISA. Elshal M.F, McCoy J.P. Methods. 2006; 38(4): 317–323.
[12] Obez Hastalarda Büyüme Hormonu, Leptin, Amilin,Glukagon Benzeri Peptid-1
Seviyeleri ile İnsülin Direnci Arasındaki İlişki [Relationship Between The Levels of
Growth Hormone, Leptin, Amylin,Glucagon Like Peptide-1 and Insulin Resistance
in Obese Patients]. Yiğitbaşı T,Baskın Y, Afacan G, Harmanda A. Turkish Journal of
Biochemistry 2010; 35 (3); 177–182.
[13] Eotaxin and Interleukin-4 Levels and Their Relation to Sperm Parameters in Infertile
Men. Yiğitbası T,Baskın Y, Afacan G, Karaarslan F, Taheri C, Aslan D. Turkiye
Klinikleri J Med Sci 2010;30 (5)1441-1445.

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 Trends in Immunolabelled and Related Techniques
Edited by Dr. Eltayb Abuelzein

ISBN 978-953-51-0570-1
Hard cover, 360 pages
Publisher InTech
Published online 27, April, 2012
Published in print edition April, 2012

The book is coined to provide a professional insight into the different trends of immunoassay and related
techniques. It encompasses 22 chapters which are grouped into two sections. The first section consists of
articles dealing with emerging uni-and-multiplex immunolabelled methods employed in the various areas of
research. The second section includes review articles which introduce the researchers to some
immunolabelled techniques which are of vital significance such as the use of the conjugates of the
Staphylococcus aureus protein "A" and the Streptococcus Spps. protein "G" in immunolabelled assay systems,
the use of bead-based assays and an overview on the laboratory assay systems. The book provides
technological innovations that are expected to provide an efficient channel for developments in
immunolabelled and related techniques. It is also most useful for researchers and post-graduate students, in
all fields, where immunolabelled techniques are applicable.

How to reference
In order to correctly reference this scholarly work, feel free to copy and paste the following:

Türkan Yiğitbaşı (2012). Multiplex Immunoassay and Bead Based Multiplex, Trends in Immunolabelled and
Related Techniques, Dr. Eltayb Abuelzein (Ed.), ISBN: 978-953-51-0570-1, InTech, Available from:
http://www.intechopen.com/books/trends-in-immunolabelled-and-related-techniques/bead-based-multiplex

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