E3S Web of Conferences 271, 01037 (2021)

https://doi.org/10.1051/e3sconf/202127101037
ICEPE 2021

The Techniques Used on the Development of COVID-19 Vaccine

Jiatong Su 1, *
1 Lalumiere school, La Port, Indiana, 46350, USA.

Abstract. The COVID-19 pandemic, also known as the coronavirus pandemic, is caused by the severe acute
respiratory syndrome coronavirus (SARS-CoV-2). It was first discovered in Wuhan, China, in December
2019 and continues until now, which becomes one of the deadliest pandemics in history and leads to a global
recession. Thus, the production of the vaccine against SARS-CoV-2 become extremely urgent for all countries
in order to control the epidemic. Therefore, the scientific community has made the rapid and significant
progress in the development of vaccines against COVID-19, i.e., the entire vaccine manufacturing and
production cycle has been greatly shortened. Up to now, more than 200 candidate vaccines have been created.
In this paper, several vaccine technologies commonly used in the manufacture of COVID-19 vaccine are
summarized. Besides, the different technologies that have been utilized for manufacturing are introduced.
Furthermore, the corresponding clinical data are listed and discussed for the sake of indicating the success or
failure of the vaccine.

1Introduction
1.2 Structure of COVID-19
1.1 Overview of COVID-19
Compared with the SARS coronavirus (a kind of virus that
Starting in 2020, all the people in the world face the crises spread in 2003 in China), COVID-19 and SARS have 80%
and the challenge of COVID-19 pandemic, which is also sequence similarity in the genes. Like SARS, COVID-19
known as the coronavirus pandemic. It has great has a layer of single-stranded RNA coated on the outside
infectiousness and causes an enormous number of of another single-stranded RNA covering by the surface
infections. Figure 1 demonstrates the complete of some spines. As a matter of fact, it is the typical
pathogenesis of COVID-19 representing all the facts structure of coronavirus, which is a linear single stranded
related to its virulence inside the host body. On this RNA virus. Besides, novel coronavirus is usually
occasion, each country is eager to find an effective spherical with irregular shape [2].
treatment and pour plenty of funding on the research of
the vaccines. 1.3 Transmission of COVID-19
COVID-19 spreads mainly through the air and when the
infected person breathe, cough, sneeze, or speak.
Moreover, the incubation period of COVID-19 is long
(about 1 to 14 days even up to 24 days), which means
people will spread the virus even without symptoms. In
addition, COVID-19 has highly variable of symptoms
showing up from none to severe illness. All these
problems cause the highly infectious characteristics of
COVID-19 and increase the difficulty of the work to limit
the speed of the virus. Until now, more than 90 million
people were diagnosed with more than 2 million
cumulative deaths. To be more specific, the U.S.A has
more than 350,000 people dead in that disaster, which is
the country with the highest number of deaths; Brazil
stayed at the second position, owned more than 200,000
people dead due to COVID-19. In order to restrain the
disease, all the countries in the world have paid the great
Figure1 Schematic representation of COVID-19 pathogenesis: financial, material and human resources. For example,
A complete profile of entry, integration and replication of virus
most of the governments block down the frontier, limit the
[1]

*Corresponding author. Email: [email protected]

© The Authors, published by EDP Sciences. This is an open access article distributed under the terms of the Creative Commons Attribution
License 4.0 (http://creativecommons.org/licenses/by/4.0/).
E3S Web of Conferences 271, 01037 (2021) https://doi.org/10.1051/e3sconf/202127101037
ICEPE 2021

transfer of citizens, looking for the cure to the COVID-19 molecules changes the coding protein, i.e., proteins are not
as well as support the development of vaccines. affected by the physical and chemical properties of
different products [5]. Thus, the vaccine platform will be
more timesaving and cost-saving. The manufacturing of
1.4 Treatment of COVID-19
mRNA avoids the lengthy cell-culture and purification
Unfortunately, we still haven't found any effective process involved in traditional viral vaccine production, as
treatment options so far. In this case, most of the methods well as the stringent requirements for biosecurity
applied in clinic treatment are comprehensive treatments. measures. Thereby, it is useful and suitable for the
Currently, with the support of respiratory, only some suppression of pandemic [6].
antiviral drugs (e.g., Redecivir, Chloroquine phosphate, In terms of efficacy, mRNA does not need to cross the
Fapilavir, etc) are effective for the treatments. As for the nuclear membrane to treat patients compared to DNA,
Redecivir, originally a drug used to treat Ebola, it is able which makes it even more effective [7]. mRNA vaccines
to improve clinical symptoms (e.g., reduce the fever). lack MHC haplotype limitations compared to
With regard to Chloroquine phosphate, an commonly used polypeptides, which is the requirement that APC or target
antiviral or anti-malarial drug, it inhibited the Novel cells express MHC molecules recognized and responded
Coronavirus infection of cells. For Fapilavir, a drug used by the T cell. In addition, mRNA vaccines can be designed
to treat influenza, it was also found to have significant as self-adjuvants (an advantage over peptide- and protein-
inhibitory effect on novel coronavirus in clinical trials. based vaccines) on account of the binding functions with
Inspiringly, the development of the vaccine already has pattern-recognition receptors.
great success with plenty of novel vaccine technology
including mRNA vaccine, inactivated vaccine, virus 2.2 Introduction of mRNA Vaccine
vector vaccine, and recombinant protein vaccine(as shown
in table1). With the development of mRNA modification and
delivery tools, the field of mRNA vaccines has progressed
Table1: Classification of COVID-19 vaccine technical route rapidly in recent years in terms of basic and clinical
DNA RNA Recombin Inactiv Virus research. Clinical trials of mRNA vaccines against viral
ation ated vector diseases (e.g., Zika, Ebola, influenza, rabies and
Protein cytomegalovirus infection) have been conducted in many
Genexine( Moderna cdbio sinovac AstraZe countries [6]. Meanwhile, as long as the selected antigen
neca
AnGes BioNTech Vaxine Immun cansinot of the pathogen target is found out, the gene sequence of
itor ech the pathogen can be detected and cloned into the DNA
Inovio Arcturus Kentucky Bharat Gamale template plasmid, which greatly increases the
BioProces Biotech ya manufacturing efficiency of mRNA vaccine. After the
sing
vaccine is injected into a subject, the mRNA vaccine uses
Cadila(CADI CureVac(CV
LAHC NS) ACO) the host cell's mechanisms to convert the mRNA into his
antigen in the body. The final cell location of antigen is
determined by signal peptide and transmembrane domain.
2 mRNA Vaccine This can be inherent to the natural protein sequence or
engineered to guide the protein into the desired cell
compartment. Thus, antigens are able to be expressed as
2.1 The introduction of mRNA
intracellular, secretory, or membrane-bound proteins [8].
The concept of mRNA as a therapeutic drug was formally Owing to the RNA replication mechanism (which
proposed in the 1990s. Subsequently, mRNA technology contains the virus gene), the structural protein sequence
gradually developed into a platform with great potential will be replaced by other genes. Therefore, the self-
for vaccine products due to the simple structure. As the replication of genes containing viruses will be guided by
smallest genetic structure, it contains only the elements RNA and multiple antigen-encoding messenger RNAs
that are directly required for protein expression. will be produced. This mimics viral infection and causes
Meanwhile, it has the good security with following the body to produce antibodies, leading to an effective
reasons. First of all, the mRNA is unable to interact with humoral and cellular immune response [9]. The host's
the genome. In addition, recombination between single- innate system is capable of sensing and responding to the
stranded RNA molecules can occur in rare cases, i.e., RNA sequences from which the virus originated, i.e., an
potentially harmful genome integrations are ruled out. mRNA vaccine will induce a strong innate response.
Moreover, mRNA is only a transient information carrier These include the production of chemokines and
owing to the not-replicable characteristic and genomic cytokines at the injection site, such as interleukin-12 (IL-
integration gaps [3, 4]. 12) and tumor necrosis factor (TNF). These are key factors
In theory, as a therapeutic molecule, mRNA could for inducing an effective adaptive response to the coding
provide breakthroughs in treatments and vaccines. Since antigen successfully [10]. In addition, mRNA is fully
mRNA can write and express all proteins, it suggests a synthetic and almost any sequence can be designed and
principle possible way to develop vaccines for the synthesized in silicon, delivered as an mRNA vaccine as
prevention and treatment of various diseases, e.g., well as quickly tested in vivo in animal models.
influenza, cancer and COVID-19. The sequence of RNA

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Although there are significant theoretical benefits to Similarly, mRmRNA-1273 is also suitable for stable
an mRNA vaccine, the fastest developing product is still perfusion. A phase 1 dose escalation open label trial
in clinical trials. The biggest obstacle is the activation of demonstrated the safety and reactivity of this vaccine and
innate immune response, which is a double-edged sword induced neutralizing antibodies after two injections. The
for mRNA vaccines. On the one hand, mRNA induces experiment involved 45 healthy adults aged from 18 to 55
immune protection through immune activation. On the years who received two mRmRNA-1273 doses of 25, 100,
other hand, excessive activation of innate immunity will or 250 μg, 28 days apart. There were 15 participants in
prevent mRNA translation and degrade mRNA. From a each dose group [16]. The safety of the vaccine was also
druggable point of view, a qualified mRNA vaccine tested in elderly people aged among 56 to 70 years and
product requires that the mRNA structure is stable enough with a BBB of 0 or above, with two doses of either 25 μg
to be delivered and expressed in target cells while or 100 μg. In this population, adverse events at both doses
inducing the desired adaptive immune response. were moderate. The vaccine also induces a CD4+ T cell
response, including production of IFN-γ, IL-2, and TNF-
α [17].
2.3 Mechanism of mRNA vaccine
So far, two phase III mRNA vaccines, BNT162B2 and 2.4 Clinical Data
MRNA-1273, have been shown to be at least 50 percent
effective [11]. BNT162B1 is a lipid nanoparticle whose A Phase 1/2/3 global clinical trial of BNT162B2 vaccine,
nucleoside modified mRNA encodes the S-receptor funded by Biotech and Pfizer, evaluated the safety,
binding domain. It has another version, which is encoded immunogenicity, and efficacy of 30 μg BNT162B2
stable, membrane-anchored full-length pre-fusion S against COVID-19 in people from 16 years and older.
BNT162B2 [12]. mRNA vaccine technology based on Results of Phase 2/3 safety and efficacy of BNT162B2
lipid nanoparticles (LNP) will deliver precise genetic vaccine were shown in the trial.
information and adjuvant effects to antigen presenting A total of 43,548 participants were randomized, with
cells S4. The efficacy of this technique has been proved 21,720 receiving the BNT162B2 vaccine and 21,728
against a variety of virus targets. RNA vaccines receiving a placebo. A total of 170 infected people were
synthesized have been shown to be safe and well tolerated negative after the second injection, of which 8 were in
in clinical trials from LNP and liposomes for the BNT162B2 recipients and 162 were in placebo recipients.
prevention of infectious diseases or the treatment of Of the 10 cases of severe COVID-19 that developed after
cancer [13]. In addition, the BNT162B1 vaccine optimizes the first injection, 9 of them were in placebo recipients and
RNA for high stability and translation efficiency. 1 was in BNT162B2 recipients. BNT162B2 was 95%
Furthermore, it has a receptor binding domain (RBD) of effective in preventing COVID-19 (95% confidence
the SARS-CoV-2 spike protein, which is a key target of interval 90.3 to 97.6). Similar vaccine potency (typically
neutralizing antibodies. The RBD antigen expressed by 90-100%) was observed in age, sex, race, ethnicity,
BNT162B1 is fused to the T4 fibrin-derived "fold" trimer baseline body mass index, and subgroups with or without
domain, which increases its immunogenicity through comorbidities. The safety profile of BNT162B2 was
multivalent display. Meanwhile, the BNT162B1 vaccine short-term, mild to moderate pain, fatigue, and headache
conjugates 1-methyl-pseuduracil to inhibit innate immune at the injection site. The incidence of serious adverse
response and increase mRNA translation in vivo [14]. events was lower and similar in the vaccine and placebo
In a phase 1/2 placebo-controlled trial, in order to groups. Therefore, one draws the conclusion that
demonstrate dose-dependent response, 45 adults were BNT162B2 two-dose regimen provides 95% protection
randomized to 2 doses of 10,30 or 100 μg. Doses were 10 against COVIAD-19 in people aged 16 years and older.
μg, 30 μg and 100 μg [15]. No serious adverse events were The median safety of 2 months was similar to that of other
recorded. Local injection site reactions and systemic viral vaccines [18].
events (mainly influenza-like symptoms) are dose- A phase 3 clinical trial of the MRA-1273 vaccine,
dependent, generally mild to moderate, and transient. The funded by the Biomedical Advanced Research and
antibodies, which recognize the receptor binding domain Development Agency as well as the National Institute of
and the neutralizing antibodies, are also activated in a Allergy and Infectious Diseases, has demonstrated an
dose-dependent manner, and increased after the second evaluation of the safety and efficacy of the MRA-1273
dose. Serum RBD binding immunoglobulin G (IgG) vaccine in preventing SARS-CoV-2 infection(table2).
concentration and SARS-CoV-2 neutralization titer also The trial enrolled 30,420 volunteers, who were randomly
increased with the augment of the dose and after the assigned in a 1:1 ratio to receive either the vaccine or a
second dose. After 14 days of dose increase, the geometric placebo (15,210 participants in each group). More than 96%
mean neutralization titer reached 1.9-4.6 times that of of participants received both injections, and 2.2% had
COVID-19 human convalescence serum (HCS). The evidence (serological, virology, or both) of SARS-CoV-2
vaccine was then compared with a second version of the infection at baseline. Symptomatic COVID-19 disease
stable, membrane-anchored full-length prefusion, was confirmed in 185 participants in the placebo group
BNT162B2. Interestingly, both vaccines produced similar (56.5/1000 person-years; 95% confidence interval [CI],
dose responses in both younger and older adults, and 48.7 to 65.3) and mRNA 1273 for 11 participants
BNT162B2 was selected for further clinical studies due to (3.3/1000 person-years; 95% confidence interval, 1.7 to
the higher safety [11]. 6.0). The efficacy of the vaccine was 94.1% (95%

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confidence interval, 89.3-96.8%; P < 0.001). In key the mRNA-1273 group. Serious adverse events were rare
secondary analyses, efficacy was similar, including and the incidence was similar between the two groups.
assessments 14 days after the first injection, analyses of There may be transient local and systemic reactions after
participants who had evidence of SARS-COV-2 infection mRNA-1273 injection. Whereas no safety concerns have
at baseline, and participants over 65 years old. 30 been found from the others. To sum up, the mRNA-1273
participants developed severe COVID-19 and 1 died, vaccine showed 94.1% efficacy in preventing COVID-19
which were all from the placebo group. Moderate and disease, including severe disease [19].
transient reactivity after vaccination was more common in
Table2: The progress of COVID-19 vaccine

Organizer name Technical route Clinical number Clinical Starting Number of

stage time people
Moderna mRNA-1273 RNA NCT0470427 Ⅲ 2020727 30000
CNBG —— Inactivated Chi CTR200034780 Ⅲ 2020/7/18 15000
sinovac —— Inactivated NCT04456595 Ⅲ 2020/7/1 8870
vaccine
AstraZeneca COV003 Virus vector ISRCTN89951424 Ⅲ 2020/5/28 2000
BioNTech BNT162b28 RNA —— Ⅲ 2020/7/27 30000
cansinotech Ad5-nCov Virus vector ChicTR2000031781 Ⅱ 2020/4/12 500
Academy of —— recombination NCT04341389 Ⅱ 2020/4/12 508
Military
Medical
Sciences

3 Inactivated whole-virus vaccines 3.2 Clinical Data

Coronavac, the COVID-19 inactivated, and aluminum
3.1 Introduction of inactivated vaccine adjuvant vaccine produced by China Kehua Biological
Products Co., Ltd., has completed a phase I/II efficacy and
An inactivated vaccine is one in which a virus or safety, double-blind, randomized, placebo-controlled
bacterium is grown and then inactivated with heat or a clinical trial [20] ( as shown in Table2) . The phase II trial
chemical usually formalin. An inactivated vaccine can be involved 600 adult participants (ages 18 to 59).
composed of whole viruses or bacteria, or of their lysed Participants were randomly assigned to one of two dual-
fragments, as a lysed vaccine. Lytic vaccines are produced dose programs, 0 to 14 days, and 0 to 28 days. In each plan,
by further purification of the microorganism until the 120 participants were given a 3 μg dose, 120 participants
vaccine contains only the desired antigenic components, were given a 6 μg dose, and 60 participants were given a
such as pneumococcal polysaccharides. placebo. Local adverse events such as pain and swelling
Inactivated vaccines often require multiple doses since were mild to moderate in the treatment regimen. No
a single dose is unable to produce protective immunity but serious grade 3 adverse events have been reported. In both
merely "initializes" the immune system. A second or third regimens, participants who were given 3 and 6 μg had high
dose is required to produce protective immunity. The NAB responses. At 28 days after the second
immune response is usually humoral immunity. Besides, administration, NAB levels in the 0- and 14-day groups
the antibodies it produces have the function of neutralizing were stable, but significantly increased in the 0 and 28 day
and eliminating pathogenic microorganisms as well as groups. A similar pattern was observed for specific
their toxins, but rarely or not causes cellular immunity. antibodies. In addition, NAB levels decreased with age,
Antibody titers produced by vaccination with inactivated suggesting that older people need to fortify the dose [21].
vaccines decrease with time, i.e., some inactivated The COVID-19 inactivated and aluminum adjuvant
vaccines require periodic booster vaccination. Inactivated vaccine Coronavac is currently in phase III clinical trials.
vaccines, which are usually unaffected by circulating It documented the occurrence of adverse events associated
antibodies and can be administered even if antibodies are with two doses of COVID-19 adsorbed vaccine
present in the blood, cannot replicate in the body and can (inactivated vaccine) produced by Sinovac within 1 week
be used in immunocompromised people. The advantage of of vaccination in adults (18-59 years old) and older adults
inactivated vaccines is that they are safe to use, easy to (60 years old or older). Its phase 3 clinical trials are being
save, no pollution risk, and insensitive to the conducted in Brazil and Indonesia due to the low number
neutralization of maternal antibodies. Which shows the of active cases in China [20].
high feasibility for the product of COVID-19 vaccines by Except for Coronavac, Sinopharm is working with the
this means. Wuhan Institute of Biological Products and the Beijing
Institute of Biological Products to develop an inactivated
vaccine. The vaccine candidate is currently in phase 3
trials [22]. According to Wuhan Biologics Research, a

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preliminary assessment for the safety and immunogenicity addition, Ad is capable of inducing apoptosis of tumor
of a candidate inactivated vaccine in healthy Chinese cells (also known as oncolytic cells). Ad has great
adults was reported after 28 days of vaccination. It potential in the development of vaccines to prevent
contains the phase I and phase II double-blind randomized infectious pathogens or cancer treatments [25]. On
clinical trial. The vaccine was given three doses in the account of its high safety, Ad is able to replicate almost all
phase I trial and two in the phase II trial. The trials were living cells and can be absorbed without adjuvants via oral,
conducted in Henan Province, China. The first phase intranasal, or intramuscular routes. At the same time, Ad
involved 96 healthy adults aged from 18 to 59. The second infection is usually mild in humans but can sometimes be
phase consisted of 224 participants of the same condition. life-threatening, especially in immunocompromised
A total of 320 randomized patients completed two individuals.
phases of the trial within 28 days of full vaccination. The The AD genomic DNA is about 26-45kb, with two
average age of the participants was 42.8 years. In the 100-140bp inverted terminal repeats at each end. The
phase 1 trial, 96 participants were assigned to one of three genes expressed in the ADS life cycle are generally
dose groups, 2.5, 5, and 10 μg/ dose, the aluminum divided into two types: early genes and late genes. Early
hydroxide (alum) adjuvant only group, and received three genes promote ADS replication by changing the
intramuscular injections on days 0, 28, and 56. High NAB expression level of host related genes. Late genes assist in
response and seroconversion were observed in all the cleavage of host cells and the assembly as well as
participants in the low-dose and high-dose groups 14 days release of virions. Ad virions are mainly composed of two
after the third vaccination (Day 70), with 23 of them proteins, the capsid protein and core protein. Core proteins
(95.8%) in the medium-dose group. In the phase 1 trial, act as DNA-related proteins, including V, VII, and X
high levels of specific antibody responses were also proteins [26]. Among them, VII protein plays a key role
produced. Besides, seroconversion was observed in all in DNA manipulation, e.g., DNA binding, initiation of
participants. The most common adverse reactions were DNA replication, protection of viral genome, etc. Capsid
pain at the injection site and self-relieving fever. proteins consist of Hexon, Penton, fiber, IIIA, VIII, and
In the phase 2 trial, 224 adults were randomly assigned IX [27]. Hexons are the main structural proteins on the
to two treatment groups, 0 and 14 days after injection, and capsid with 240 hexagonal trimers located on the surface
0 and 21 days after injection. In each treatment group, 84 of Ad virions. There are several hypervariable regions on
people were assigned to the medium-dose (5 μg) vaccine the Hexons, which are the main neutralization sites of Ad
group and 28 to the aluminum adjuvant placebo group. and can be replaced by other foreign antigens as potential
Five patients (6%) in the 5 μg group and 4 patients (14.3%) vaccine vectors. The IIIa protein is located on the surface
in the placebo group experienced adverse reactions during of the undergarment and Ad in the proper assembly of the
0 and 14 days of treatment. 16 patients (19%) in the 5 μg virus, the stabilization of the vertex region, and the
group and 5 patients (17.9%) in the placebo group had assembly of the encapsulating genome. VI protein is in the
adverse events during the 0 and 28 days of treatment. Both capsid and serves as the key cleavage factor of AdS in the
treatments had higher NAB response and the serum process of endosomal destruction. The VIII protein
conversion rate was 97.6%. In addition, for specific provides the bonding between the surrounding pentagonal
antibody responses, 0 - and 21-day antibody responses prism and hexahedrons, maintaining the stability of the
were much higher than 0 - and 14-day antibody cases. capsid. The function of IX protein is to inhibit the innate
Seroconversion was also relatively low at 0 - and 14-day immune response and maintain the virality and stability of
treatments, at 85.7%, compared with 100% at 0 - and 21- the capsid [28].
day treatments. Adverse reactions, as in the phase 1 trial,
were injection site pain and self-relieving fever [23]. In
4.2 Adenovirus Vector Vaccine
this interim report on the Phase I and Phase II trials of
COVID-19 inactivated vaccine, patients had a low Ad vector is one of the most effective vectors for the
adverse event rate and showed immunogenicity, which delivery of foreign antigen to host cells. Therefore, the
indicates great potential for development. strategy of viral vector vaccination has obvious
advantages. Besides, among the virus vectors under study,
4 Adenovirus Vector Vaccine adenovirus (AdV) is the most promising vaccine vector.
As a vector for gene therapy, Adenoviral-Vec vaccines
have the inherent ability to deliver genes to mammalian
4.1 Introduction of adenovirus cells while Adv is the vector for live vaccines [29]. As a
popular vaccine vector, AdV has high safety, high gene
Adenovirus (Ad) is an unencapsulated DNA virus with an transfer potential, broad cell orientation, and well-
icosahedral capsid and a diameter of about 90 nm. Rowe characterized genomes as well as their operational
and his colleagues first discovered this in 1953, during an protocols [30].
experiment to grow adenoid tissue [24]. Subsequently, in AdV stands out for its safety as a vaccine. Early in the
1993, the first ADS-based study of human gene therapy 1970s, the U.S. Army developed the AdV Serotype 4
was performed with the first in vivo gene therapy in a 23- (HAdV-4) and HAdV-7 vaccines for export to prevent
year-old homozygous cystic fibrosis patient. He was given acute respiratory disease caused by ADHU4 and ADHU7
the E1-E3 deletion RAD vector of normal human CFTR. with no side effects [31]. Meanwhile, the AdV vector has
In the 2000s, this technology was widely applied since high immunogenicity. They can transduce APCs and
Ads could cause the reaction of T cells and B cells. In

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induce humoral and cellular immune responses, which NIHR Clinical Research Network, and AstraZeneca, to
may be enhanced by the initial induction of a strong innate document the safety and immunogenicity of ChAdOx1
response. First, after APCs ingest virions, intracellular nCoV-19 vaccine in young and older adults.
viral proteins are processed and presented to CD8+ T cells There were 560 participants in the study. Of the 160
along with major histocompatibility complex (MHC) participants aged from 18-55 years, 100 were assigned to
class I molecules. This leads to the production of Ad- Chadox1 nCOV-19 and 60 to Menacwy. Among 160
specific cytotoxic T lymphocytes (CTL), which clear AD- participants aged from 56 to 69 years, 120 were assigned
transduced cells. Furthermore, MHC molecules presented to Chadox1 nCoV-19 and 40 to Menacwy. With regard to
with de novo synthesis of viral proteins lead to type 1 240 participants over 70 years of age, 200 were assigned
activation of CD4 + T-helper (Th1 cells). Th1 cells secrete to Chadox1 nCOV-19 and 40 were assigned to Menacwy.
cytokines, such as interleukin-2 (IL-2) and interferon-γ A total of 552 participants were analyzed, of which 280
(IFN-γ), induce further proliferation of AD-specific CTLs. (50%) were female. Local and systemic reactions were
Moreover, IFN-γ can up-regulate the expression of MHC more common in participants who received the Chadox1
class I molecules in Ad-transduced cells and promote their Novel Coronavirus-19 compared to those who received
recognition by CTL. AdV enters B cells and is the control vaccine. The presenting of them contain
subsequently processed to input virion coat proteins, injection site pain, fever, muscle pain, and headache, but
which are present on the surface of B cells along with were less common in the elderly (age ≥56 years) than in
MHC class II molecules, activating Th2 cells to secrete younger participants. In those who received two standard
cytokines such as IL-4, IL-5, IL-6, and IL-10. These doses of Chadox1 NCOV-19, post-vaccination local
cytokines induce the differentiation of these B cells into reactions were reported in 43 (88%) participants in the 18-
Ad specific antibody secretory plasma cells. Overall, the 55 years group,22(73%) in the 56-69 years group and
inherent immunogenicity of AdV makes it an attractive 49(61%) in the elderly population aged 70 years and older,
vaccine vector. Nevertheless, it also creates barriers to its in 42(86%) and systemic reactions in participants in the
usage. In detail, Anti-AdV immunity, whether pre- 18-55 years group, 23(77%) in the 56-69 years group and
existing or caused by the AdV vaccine, may impair 32 70 years and older group (65%). As of 26 October 2020,
vaccine effectiveness [32]. there have been no serious adverse events related to the
investigational vaccine.
Among participants who received two doses of
4.3 Clinical Data
vaccine, the median 28-day anti-spike SARS-CoV-2 IgG
A recombinant adenovirus type 5 vector Covid-19 vaccine, response was similar across three age groups (standard
funded by the National Key R&D Programme of China, dose group :18-55 years, 20 713 arbitrary units [AU]/mL
the National Science and Technology Major Project, and [IQR 13 898-33 550], n=39; 56-69, 16 170 Au /mL [10
Cansino Biologics, has been tested in phase 2 233-40 353], n=26; ≥70 years 17 561 Au /mL [9705-37
immunogenicity and safety trials in healthy adults aged 18 796], n=47; P = 0 ꞏ 68). Neutralizing antibody titers after
years or older. the enhanced dose were similar in all age groups (median
The randomized, double-blind, placebo-controlled MNA80 on day 42 in the standard dose group :18 -- 55
clinical trial was conducted at a center in Wuhan, China. years, 193 [IQR 113-238], n=39; 56-69 years old, 144
Participants were all healthy adults over18 years old with [119-347], n=20; ≥70 years old, 161 [73-323], n=47; P =
a total of 508 participants. The mean age of participants 0 ꞏ 40). After 14 days of booster administration, 208 of
was 39ꞏ7 years (SD: 12ꞏ5; Age range 18-83), 309 (61%) 209 enhanced participants (>99%) had a neutralizing
18-44 years old, 134 (26%) 45-54 years old, and 65 (13%) antibody response. T cell response reached the peak on
55 years old and older. 253 were randomly assigned to the day 14 after a single standard dose of Chadox1 nCoV-19
1 × 1011 particle dose group, 129 to the 5 × 1010 particle (18--55 years: median 1187 speckle-forming cells per
dose group, and 126 to the placebo group. Among 129 million peripheral blood mononuclear cells [IQR 841-
subjects in the 1 × 1011 and 5 × 1010 particle dose groups, 2428], n=24; 56-69 years :797 square feet [383-1817],
227 (90%, 95% CI 85-93) and 113 (88%, 81-92) IFN-γ n=29; ≥70 years :977 SFCS [458-1914], n=48). These
ELISA responses were observed after vaccination. results denoted that ChAdOx1 nCoV-19 vaccine appeared
Adverse events were reported in 183 (72%) of 253 to be better tolerated in older adults than in younger adults
patients and 96 (74%) of 129 patients in the 1 × 1011 and and had similar immunogenicity in all age groups
5 × 1010 particle dose groups. Severe adverse reactions increased doses. Further evaluation of the effectiveness of
occurred in 24 cases (9%) of the 1 × 1011 virus particle the vaccine is still needed [34].
group and 1 case (1%) of the 5 × 1010 virus particle group.
The results showed that the 5×1010 particle Ad5 vector 5 Recombinant protein vaccine
Covid-19 vaccine was safe and induced a significant
immune response in most recipients after a single dose of
immunization [33]. 5.1 Introduction of recombinant protein vaccine
In addition, a phase 2/3 controlled trial conducted by
U. K. Research and Innovation, National Institutes for Recombinant protein vaccine is a kind of virus target
Health Research (NIHR), Coalition for Epidemic antigen gene constructed on the expression vector. The
Prepared Innovations, NIHR Oxford Biomedical constructed expression protein vector is transformed into
Research Centre, Thames Valley and South Midlands bacteria, yeast or mammal or insect cells, under certain
induction conditions, which expresses many antigen

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proteins through the purification of vaccine preparation. receive the adjuvant vaccine, 25 to not receive the
The cost of producing and purifying immunogens is high. adjuvant vaccine, and 23 to receive a placebo. All the
Besides, the production of large quantities of subjects developed IgG antibodies against spines after a
immunogenic proteins with sufficient purity after single vaccination and many of them also produced a
identification of immunogenic proteins is also an neutralizing antibody response against the wild-type virus.
important part of vaccine manufacturing. The advent of After the second dose, 100% of subjects developed a
recombinant DNA technology means that foreign genes neutralizing antibody response. Two doses of 5 g vaccine
can be inserted into expression vectors and then produced a mean geometric titer (GMT) of neutralizing
introduced into cells to aid in the production of foreign antibodies of about 3900, four times higher than in
proteins. In many cases, this provides a vast and patients recovering from COVID-19. Most participants
inexpensive source of protein for vaccination research were nonreactive or mildly reactive and the duration of
[35]. adverse reactions was short (mean, ≤2 days). One
As for bacterial expression, it uses Escherichia coli to participant had a mild fever that lasted 1 day. In addition,
establish the first recombinant expression system. This the researchers examined the participants' CD4-positive T
expression system can provide a relatively large number cell responses. The results illustrated that NVX CoV2373
of defined proteins. However, expressed proteins are often will induce antigen specific multifunctional CD4 positive
misfolded because prokaryotic cells have different T cell response. Moreover, the immune response is
processing and transport mechanisms. One of the first strongly biased towards Th1 phenotype (IFN-γ, IL-2, and
successfully produced recombinant veterinary vaccines TNF-α). The activation of CD4 positive T cells is
was based on the gp70 surface glycoprotein or p45 protein conducive to the long-term production of highly effective
of FeLV expressed in E. coli [36]. With regard to yeast neutralizing antibodies, which indicates promising results
expression, which is an industrial microorganism, the for the generation of long-term immunity. The results
widespread use of Saccharomyces cerevisiae (Baker's proved that NVX-COV2373 appeared to be safe at day 35,
yeast) makes it the alternative antigen protein expression triggering an immune response that exceeded the level of
system of choice. It makes the recombinant protein more COVID-19 recovery serum. CD4+ T cell response
likely to fold properly than the prokaryotic system. induced by matrix - M1 adjuvant was biased towards the
Moreover, the expression system of insect cells was Th1 phenotype [40].
established by Spodoptera frugiperda infected with a
baculovirus vector, Autographa californica nuclear
polyhedrosis virus [37]. In addition, mammalian cell 6 Conclusion
expression, due to the infection and replication of many Recently, positive results have been reported from Phase
veterinary pathogens in cultured mammalian cells [38]. III clinical trials of the new coronavirus in China, the
Finally, plant cell expression, an additional emerging United States, Russia, the United Kingdom and other
expression system worthy of mention. The first approved countries. Due to the great success of the research and
vaccine for the expression system is a vaccine against development of the new coronavirus on different technical
Newcastle disease (NDV) infection in poultry. It is being routes, many countries around the world have begun to
studied for other vaccine applications, including carry out large-scale vaccination work. Globally, orders
infectious bronchitis virus, infectious bursal disease virus, for COVID-19 vaccines total more than 10 billion doses,
ETEC, BVD and bovine herpes virus [39]. according to national press data.
In terms of vaccine capacity, the United States has the
5.2 Clinical Data highest capacity. Novavax's vaccines, Pfizer's, Modena's
and Johnson & Johnson's have a capacity of at least 4.8
A phase I-II trial of SARS-CoV-2 polypeptide Spike billion doses. Orders for the four vaccines totaled 6 billion
Protein Nanoparticle Vaccine, NVX-COV2373, designed yuan. The UK's AstraZeneca vaccine is also seen as a
by Novavax and funded by the Coalition for Epidemic promising alternative to the US vaccine, with orders
Prepared Innovations, was completed. NVX CoV2373 is totaling more than 2 billion doses. In addition, Russia has
a candidate vaccine engineered from a novel coronavirus received orders for 1.2 billion doses of vaccines from
gene. It was produced using Novavax's recombinant more than 50 countries, according to data released by the
nanoparticle technology to generate antigens for Novel country. In the end, China laid out a total of five technical
Coronavirus spikes. It also contains Novavax's proprietary routes to develop the new coronavirus. The total vaccine
saponin-based Matrix-M adjuvant, which enhances the capacity of China mRNA new coronavirus vaccine,
immune response and stimulates high levels of Kangtai Biotech's adenovirus technology vaccine,
neutralizing antibody production. To be more specific, it Zhifang Biotech's phase 2 clinical recombinant protein
stimulates the entry of antigen-presenting cells into the new coronavirus vaccine, and the inactivated vaccine
injection site and enhances antigen presentation in local produced by Sinopharmaceutical Beijing Co., Ltd.,
lymph nodes. Sinopharmaceutical Wuhan Co., Ltd., and Beijing
In Phase 1 clinical trials, a total of 131 adults aged Kexingzhongwei Co., Ltd., will exceed 2.2 billion doses.
from 18 to 59 years received two vaccine injections to And the total number of vaccine orders in China is 500
evaluate the safety and immunogenicity of rSARS-CoV-2 million.
vaccine. Participants were randomly assigned to receive This is a great blessing for the society, which is now in
the adjuvant vaccine, with 83 participants assigned to a state of great depression. After the mass production and

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