REVIEW

published: 14 July 2022

doi: 10.3389/fmicb.2022.927306

A Comprehensive Review of the

Protein Subunit Vaccines Against
COVID-19
Mohsen Heidary 1 , Vahab Hassan Kaviar 2 , Maryam Shirani 3 , Roya Ghanavati 4 ,
Moloudsadat Motahar 5 , Mohammad Sholeh 6 , Hossein Ghahramanpour 7 and
Saeed Khoshnood 2*
1
Cellular and Molecular Research Center, Sabzevar University of Medical Sciences, Sabzevar, Iran, 2 Clinical Microbiology
Research Center, Ilam University of Medical Sciences, Ilam, Iran, 3 Toxicology Research Center, Medical Basic Sciences
Research Institute, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran, 4 School of Paramedical Sciences,
Behbahan Faculty of Medical Sciences, Behbahan, Iran, 5 Department of Microbiology, School of Medicine, Ahvaz
Jundishapur University of Medical Sciences, Ahvaz, Iran, 6 Department of Microbiology, Pasteur Institute of Iran, Tehran, Iran,
7
Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran

Two years after severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), in

Edited by: December 2019, the first infections were identified in Wuhan city of China. SARS-CoV-
Seil Kim, 2 infection caused a global pandemic and accordingly, 5.41 million deaths worldwide.
Korea Research Institute of Standards
Hence, developing a safe and efficient vaccine for coronavirus disease 2019 (COVID-19)
and Science, South Korea
seems to be an urgent need. Attempts to produce efficient vaccines inexhaustibly are
Reviewed by:
Dharmendra Maurya, ongoing. At present time, according to the COVID-19 vaccine tracker and landscape
Bhabha Atomic Research Centre provided by World Health Organization (WHO), there are 161 vaccine candidates in
(BARC), India
Ran Wang, different clinical phases all over the world. In between, protein subunit vaccines are types
Capital Medical University, China of vaccines that contain a viral protein like spike protein or its segment as the antigen
*Correspondence: assumed to elicit humoral and cellular immunity and good protective effects. Previously,
Saeed Khoshnood
this technology of vaccine manufacturing was used in a recombinant influenza vaccine
[email protected]
(RIV4). In the present work, we review protein subunit vaccines passing their phase
Specialty section: 3 and 4 clinical trials, population participated in these trials, vaccines manufactures,
This article was submitted to
Virology,
vaccines efficiency and their side effects, and other features of these vaccines.
a section of the journal
Keywords: SARS-CoV-2, vaccine, COVID-19, protein subunit, review
Frontiers in Microbiology
Received: 24 April 2022
Accepted: 08 June 2022 INTRODUCTION
Published: 14 July 2022
Citation: The world has been fighting coronavirus disease 2019 (COVID-19) illness for nearly 2 years
Heidary M, Kaviar VH, Shirani M, (Koupaei et al., 2022a; Naimi et al., 2022). COVID-19 was induced by severe acute respiratory
Ghanavati R, Motahar M, Sholeh M, syndrome coronavirus 2 (SARS-COV-2), which emerged in Wuhan, China (Koupaei et al., 2021;
Ghahramanpour H and Khoshnood S
Mahdizade Ari et al., 2022). Traditional vaccine strategies developed on the basis on attenuated or
(2022) A Comprehensive Review of
the Protein Subunit Vaccines Against
inactivated pathogens have been demonstrated to be highly efficient for several infectious diseases
COVID-19. (Khoshnood et al., 2022). However, in some situations, the whole pathogen method cannot provide
Front. Microbiol. 13:927306. the needed protection without adverse reactions viz inflammation, allergic, and autoimmune
doi: 10.3389/fmicb.2022.927306 responses (Baird and Lopata, 2014).

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Heidary et al. Protein Subunit Vaccines Against COVID-19

Subunit vaccines based on microorganisms’ fragments have vs. unadjuvanted groups, in high dose vs. low dose groups, and
the ability to overcome these challenges. Subunit vaccines contain younger vs. older adults (Pasteur, 2020).
only include the pathogens antigenic components that are Sanofi-GSK began phase 2 trial with 722 individuals in the
required to elicit effective immune responses. A polysaccharide, United States and Honduras in February 2021. This trial included
a nucleic acid or a protein can all be used as antigen (Bill, 2015). healthy adults aged 18 and above to assess the immunogenicity,
Protein subunit that contains a specific product of the virus safety, and reactogenicity of two injections given 21 days
rather than complete viral particle is used to elicit immune apart at three antigen dosage levels of 5, 10, and 15 µg
responses. In addition to accessory proteins, SARS-CoV-2 (Pasteur, 2021).
comprises of structural and non-structural proteins. S, membrane The interim results of the phase 2 study showed that the
(M), and envelope (E) proteins are the main structural proteins of adjuvanted recombinant COVID-19 vaccine candidate produced
SARS-CoV-2 (Koupaei et al., 2022b). These proteins are situated significant rates of neutralizing antibody responses in all adult
in the viral phospholipid bilayer and in the nucleocapsid (N) age categories, with seroconversion rates ranging from 95 to
protein, the ribonucleoprotein core (Figure 1). The S-proteins by 100%. High neutralizing antibody levels were also made after a
binding to ACE2 (angiotensin-converting enzyme-2), facilitate single injection in individuals with signs of previous SARS-CoV-
host cell attachment and viral entry. Protein S consists of two 2 infection, indicating that the vaccine has a lot of promise for
subunits, S1 and S2. The S1 subunit is a C-terminal receptor- development as a booster vaccine (Sanofi, 2021b).
binding domain (RBD) that detects the receptor, and the S2 Sanofi and GSK began enrolling patients in their Phase 3
subunit is used for membrane fusion, which is required to enter clinical study (VAT00008) in May 2021 to assess their adjuvanted
the host cell. M proteins play a role in the formation of virion recombinant protein’s safety, efficacy, and immunogenicity
envelope. Antibodies neutralize the virus by binding to the S COVID-19 vaccine candidate. More than 35,000 people aged
protein, thus making the spike gene sequence or protein a key 18 and above from different countries take part in the global,
component of SARS-CoV-2 vaccines. E proteins are important randomized, double-blind, placebo-controlled Phase 3 study,
for SARS-CoV-2 infectivity and N-protein binds the viral RNA which takes place in the United States, Asia, Africa, and Latin
genome and forms the helical (Currier et al., 2021). America. Participants were categorized as naïve (not previously
S protein of SARS-CoV-2 has been shown, to be an ideal target infected) or non-nave (previously infected) based on evidence of
for vaccine development on multiple platforms due to its high the previous disease. The overall number of participants in the
antigenicity and potency to induce robust immune responses study to be about 37,000. The experiment split into two stages.
(Figure 2; Martínez-Flores et al., 2021). However, since only a The first stage consists of a monovalent vaccination carrying the
few viral components are included in the protein subunit vaccine original SARS-CoV-2 spike protein (D614). The second stage
that do not exhibit the full complexity of the virus antigen, their consists of a bivalent vaccine having both the D614 spike protein
protective effect may be limited and, in some cases, may elicit and spike protein of the B.1.351 variant (Sanofi, 2021a). This
unbalanced immune responses (Dong et al., 2020). study, NCT04537208, is registered with ClinicalTrials.gov and is
This article reviews the protein subunit SARS-CoV-2 vaccine currently underway. No findings have been published as of yet
approaches (Table 1) with a special focus on the efforts currently since phase 3 is still in progress (Sanofi, 2021c).
underway in the development of protein subunit SARS-CoV-2 In parallel with the phase 3 study, Sanofi Pasteur has launched
vaccines. Here, we describe the all studies of the protein subunit an extension of the phase 2 booster study (NCT04762680) to
SARS-CoV-2 vaccine and demonstrate their immunogenicity, evaluate different formulations of the candidate vaccine as a
efficacy, and safety. primary series and as a booster dose against COVID-19 in
over 4,500 adults who have previously been vaccinated (Sanofi,
2021d). As previously stated, no data from phase 3 and phase 2
RECOMBINANT SPIKE PROTEIN boosters was released.
VACCINES
SCB-2019 Vaccine
VAT00002 Vaccine Clover Biopharmaceuticals (Chengdu, China) developed SCB-
Sanofi Pasteur and GSK developed the VAT00002 Sanofi–GSK 2019, a protein subunit COVID-19 vaccine that combines
COVID-19 vaccine, a recombinant protein subunit vaccine a trimeric version of the SARS-CoV-2 spike protein (S-
containing the SARS-CoV-2 spike protein generated in insect Trimer) with one of two adjuvants: AS03 (GlaxoSmithKline)
cells using a baculovirus vector. Sanofi provides the recombinant or CpG/Alum (Dynavax) (Liu et al., 2017). The spike protein
antigen, while GSK (GlaxoSmithKline, Belgium) provides the is presented in its natural three-part form in the vaccine,
adjuvant (AS03), which helps the immune system response resulting in a more effective immune response. S-Trimer is a
(Garçon et al., 2012; Goepfert et al., 2021). recombinant SARS-CoV-2 fusion protein produced in Chinese
To identify a formulation, antigen dose, and a dosing hamster ovary cells using Trimer-Tag technology. It maintains
schedule for further development of CoV2 preS dTM vaccine S-natural protein’s trimeric structure in the prefusion state of the
candidate, Sanofi-GSK began Phase 1 studies (VAT00001) in the antigenic epitope, which is required for viral neutralization, and
United States in September 2020 with 439 healthy adults aged it binds to human ACE2 with great affinity (Liang et al., 2021).
18 and above. This trial showed that Neutralizing and binding According to pre-clinical studies, S-Trimer adjuvanted with
antibodies after two doses of vaccine were higher in adjuvanted either AS03 or CpG 1018 with alum may elicit potent humoral

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Heidary et al. Protein Subunit Vaccines Against COVID-19

FIGURE 1 | Structural proteins of SARS-CoV-2 virion. Nucleocapsid (N) protein is associated to the genomic RNA and S glycoprotein/spike (S), membrane (M), and
envelope (E) proteins, which are located in the viral phospholipid bilayer. Protein S consists of two subunits, S1 and S2. The S1 subunit is a C-terminal
receptor-binding domain (RBD) that detects the receptor, and the S2 subunit is used for membrane fusion, which is required to enter the host cell.

FIGURE 2 | Type of structures protein subunit vaccines against SARS-CoV-2 and development of T-cell and B-cell immunity by activation of antigen-presenting cells
(APCs).

and cellular immune responses in various animal species with either AS03 or CpG/Alum adjuvants, induced strong
and protective immunity against SARS-CoV-2 infection in humoral and cellular immune responses against SARS-CoV-2,
non-human primates (NHP), with no indications of disease with high virus-neutralizing activity. The above results concluded
enhancement (Liang et al., 2021). that both adjuvanted vaccine formulations are well tolerated and
In June 2020, a Phase 1 randomized, double-blind, placebo- suitable for future clinical trials (Richmond P. et al., 2021).
controlled trial clinical research in healthy adult volunteers in Richmond P. C. et al. (2021) investigated the durability of
Australia with two different adjuvants, AS03 and CpG/Alum, was antibodies in those who participated in phase 1 for up to 6 months
launched (NCT04405908). Interim findings indicated that the after vaccination. SCB-2019 produced dose-dependent immune
SCB-2019 vaccine, which included S-Trimer protein synthesized responses against wild-type SARS-CoV-2 that persisted until

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Frontiers in Microbiology | www.frontiersin.org

Heidary et al.
TABLE 1 | Protein-based vaccine against COVID-19 in clinical trials.

Vaccine name Developer Route of Clinical Type of subunit and structure Type of Efficacy Side effects References
administration/ stage adjuvant
Dose

Recombinant
spike protein
vaccines
VAT00002/ Sanofi Pasteur (French) and IM/2 Phase 3 Recombinant spike protein AS03 NR NR Kandimalla
VAT00008/ GSK (United Kingdom) (Ongoing) [1. monovalent vaccine comprising et al., 2021
Vidprevtyn spike protein D614 variant
2. bivalent vaccine comprising
spike protein of D614 and Beta
variant (B.1.351)]
SCB-2019 CloverBiopharmaceuticals IM/2 Phase 3 Recombinant trimeric spike protein AS03 (GSK) or 67.2% overall Pain at the injection site, Richmond P.
Inc. (China)/GSK)/Dynavax (data CpG/Alum efficacy against any headache, fatigue, fever and et al., 2021;
(United States) reported) severity, 83.7% myalgia Bravo et al.,
against 2022
moderateto-severe
and 100% against
severe COVID-19
COVAX-19 Vaxine Pty Ltd. (Australia) IM/2 Phase 3 Recombinant spike protein Advax-SM NR NR Chavda et al.,
(ongoing) 2021
Nanocovax Nanogen Pharmaceutical IM/2 Phase 3 Recombinant spike protein Al(OH)3 NR Pain at the injection site, Hosier et al.,
Biotechnology JSC. in (ongoing) fatigue, fever, headache, 2020; Jacob
4

(Vietnam) cough, sore throat in some et al., 2020

volunteers and one patient
exhibited serious sepsis
Razi Cov Pars Razi Vaccine and Serum IM/2 and IN/1 Phase 3 Recombinant spike protein NR NR Headache, mild fever and Pilicheva and
Research Institute (Iran) (ongoing) injection site pain Boyuklieva,
2021
MVC-COV1901 Medigen Vaccine Biologics IM/2 Phase 3 Recombinant spike protein (S-2P) CpG 1018 and NR Pain at the injection site, Hsieh et al.,
Corporation (ongoing) Al(OH)3 malaise, fatigue and fever was 2021
(Taiwan)/NIAID/Dynavax rarely reported (based on phase
(United States) 2).
EPIVACCORONA Vektor State Research IM/2 Phase 3 Chemically synthesized peptide Al(OH)3 79% Local pain at the injection site Ryzhikov A.
VACCINE (EVCV) Center of Virology and (ongoing) immunogens of the S protein of et al., 2021
Biotechnology, Koltsovo, SARS-CoV-2 conjugated to a
(Russia) carrier protein

Protein Subunit Vaccines Against COVID-19

SCTV01C Sinocelltech Ltd. IM/2 Phase 2/3 Recombinant bivalent trimeric S SCT-VA02B NR NR Eroglu et al.,
July 2022 | Volume 13 | Article 927306

(ongoing) protein 2021

Recombinant
RBD vaccines
ZF2001 Anhui Zhifei Longcom IM/2 or 3 Phase 3 Tandem-repeat dimeric RBD Al(OH)3 NR Injection-site pain, redness, Yang et al.,
Biopharmaceutical/Institute (ongoing) protein swelling at injection site and low 2021
of Microbiology, Chinese systemic adverse reactions
Academy of Sciences such as fever, fatigue,
(China) headache, nausea, Cough and
muscle pain (based on phase 2)
(Continued)
Frontiers in Microbiology | www.frontiersin.org

Heidary et al.
TABLE 1 | (Continued)

Vaccine name Developer Route of Clinical Type of subunit and structure Type of Efficacy Side effects References
administration/ stage adjuvant
Dose

Abdala/CIGB-66 Center for Genetic IM/3 Phase 3 Monomeric RBD subunit Al(OH)3 92.28% Severe adverse events were Aguilar-Guerra
Engineering and (ongoing) not reported et al., 2021;
Biotechnology (CIGB) Hernandez-
Bernal et al.,
2021;
Reardon, 2021
NEGVAC/BioE/ Biological E. Limited IM/2 Phase 3 Recombinant RBD Al(OH)3 and NR NR Verma, 2021
Corbevax/Becov2 (ongoing) CpG 1018
FINLAY-FR- Instituto Finlay de Vacunas IM/2 Phase 3 Conjugated vaccine (RBD and TT) Al(OH)3 71%after two Injection site events and fever Toledo-
2/Soberana (Cuba) (data doses and 92.4% Romani et al.,
02 reported) after booster dose 2021
UB-612 COVAXX IM/2 Phase 2/3 Multitope peptide based on Aluminum NR NR Hasanzadeh
(United States)/United (ongoing) S1-RBD-protein phosphate et al., 2021
Biomedical Inc. Asia
5

(Taiwan)
ReCOV Jiangsu Rec-Biotechnology IM/2 Phase 3 Recombinant trimeric BFA03 NR NR Biorender,
Co., Ltd. (ongoing) two-component spike N-terminal 2021b;
domain (NTD) and RBD Clinicaltrials
Gov, 2022
Nanoparticle
vaccines
Novavax/NVX- Novavax (United States) IM/2 Phase 3 Full length recombinant S Matrix M 89.7% (based on Injection-site tenderness or Dunkle et al.,
CoV2373 (data protein-micelle nanoparticle phase 3 in the pain, headache, muscle pain, 2021; Heath
reported) United Kingdom) myalgia, malaise and fatigue et al., 2021
and 92.6% (based (based on phase 3)
on phase 3 in the
United States and

Protein Subunit Vaccines Against COVID-19

Mexico)
GBP510 SK Bioscience Co., Ltd. IM/2 Phase 3 Self-assembled two- component AS03 NR NR Philippidis,
July 2022 | Volume 13 | Article 927306

(South Korea) and GSK (ongoing) nanoparticle vaccine displaying 2021

(United Kingdom) RBD of spike
NR, not reported; IM, intramuscular; IN, intranasal; Al(OH)3, aluminum hydroxide; TT, tetanus toxoid.
Heidary et al. Protein Subunit Vaccines Against COVID-19

Day 184, according to the results of this study. Three of the be evaluated in 16,876 adult volunteers. Subjects get two doses
most frequent variant types of coronavirus, including Alpha of Spikogen vaccine (25 µg) with Advax-SM adjuvant (15 mg)
(B.1.1.7), Gamma (P.1), and Beta (B.1.351), were cross-reactive or placebo, 21 days apart, in a 3:1 ratio. Furthermore, due to the
with neutralizing antibodies (Richmond P. C. et al., 2021). prevalence of the Delta variant in Iran, the efficacy of the vaccine
Following the positive results of the phase 1 trial, Clover against this variant is being assessed as part of this research
company moved forward with SPECTRA, a global pivotal (Cinnagen, 2021). According to results of clinical studies on this
phase 2/3 clinical trial evaluating the efficacy, safety, and vaccine, CinnaGen reported that Spikogen has produced a robust
immunogenicity of SCB-2019 (CpG 1018/Alum) in 29,000 immune response in 87% of recipients (Khademi et al., 2018).
adult and elderly participants from five countries across Asia, This phase is continuing, and no findings from these trials have
Latin America, Europe, and Africa (Clover Biopharmaceuticals, been published. On October 2021, Iran has approved the vaccine
2021a). for emergency use (Peeridogaheh et al., 2013).
Based on Phase 3 results announced by SCB-2019 in
September 2021, the studied vaccine showed 67.2% efficacy Nanocovax Vaccine
against all COVID-19 cases of any severity, 84% efficacy against Nanocovax is a protein subunit vaccine produced by Nanogen
moderate-to-severe COVID-19, and 100% efficacy against severe Pharmaceutical Biotechnology JSC in Vietnam. The vaccine
COVID-19 with hospitalization. Furthermore, vaccine efficacy is based on SARS-CoV-2 recombinant spike protein with
against the highly transmissible delta form, which is shared aluminum hydroxide as adjuvant. Testing the vaccine in animal
worldwide, was 79%. In contrast, vaccine effectiveness against the models has exhibited its capability to create high anti-spike
mu variant, which accounted for one-quarter of strain-identified antibody levels. Nanocovax has also been demonstrated to have
illnesses, was 59% (Clover Biopharmaceuticals, 2021b). ability to neutralize anti-spike IgG antibodies and also antibody
titers against the Wuhan variant.
COVAX-19 Vaccine The phase 2 trial of the vaccine were conducted at Military
COVAX-19 (or SpikoGen) is a COVID-19 vaccine candidate Medical Academy and the Pasteur Institute in Hanoi and
based on recombinant spike proteins developed by Vaxine Pty Ho Chi Minh, respectively. Pain was the local adverse effect
Ltd., a South Australian biotech firm. The Advax-SM adjuvant is observed after the first and the second doses, whereas fatigue
combined with a recombinant protein antigen in this vaccination. and headache were systemic effects found after each injection.
According computer models of the spike protein and human However, most AVs were grade 1 and often resolved within 7
ACE2 receptor that used by Vaxine pty Ltd., it was founded that days. Hyperglycemia and leukocytosis have been reported, as
this vaccine does reduce not only COVID-19 disease but also well. Considering these AVs, none of the phases of vaccination
blocks virus shedding and transmission (Chavda et al., 2021). were paused (Jacob et al., 2020). The results of phases 1 and
In a pre-clinical investigation, the Covax-19 vaccine elicits II trials revealed that the vaccine has an excellent safety profile
robust anti-spike antibody and T cell responses in mice. irrespective of the dose. In the phase 3 trial, T-cell responses
It protects ferrets against SARS-CoV-2 infection when given and vaccine efficiency will be examined (Hosier et al., 2020).
two consecutive intramuscular doses several weeks apart The Nanocovax has also been indicated to induce high levels
(Li et al., 2021). of S protein-specific IgG and neutralize antibody in BALB/c
Phase 1 clinical trial for the COVAX-19 vaccine started in June mice, Syrian hamsters, and Macaca leonina. Moreover, the
2020, with 40 healthy adults in Australia randomly allocated to vaccine protected the upper respiratory tract from SARS-CoV-
active vaccination (30 participants) or placebo (10 participants). 2 infection in a hamster model. The vaccine also showed
(NCT04453852). Initial findings of this Phase study revealed no adverse effects on Albino Swiss mice, brown rats, and
that the COVAX-19- the vaccine was safe, well-tolerated, and New Zealand rabbits. These pre-clinical results reflected that the
immunogenic in the participants; however, information about Nanocovax vaccine is highly safe, effective, and immunogenic
the results of this experiment has not been published yet (Khan (Deeks et al., 2020).
et al., 2020; Arashkia et al., 2021). COVAX-19 vaccine entered
into phases 2 and 3 clinical trials with a cooperation agreement Razi Cov Pars
with CinnaGen Company in Iran (Chavda et al., 2021). The Iranian-made COVID-19 vaccine, Razi Cov Pars, is a protein
In the Phase 2 trial, 400 Iranians participant were allocated to subunit vaccine comprising coronavirus-like spike proteins
receive two doses of either the active vaccine or saline placebo, (Kumar and Kumar, 2021). This first injectable inhaled SARS-
21 days apart, in a 3:1 ratio. This study aimed to assess covax- CoV-2 vaccine was developed by Razi Vaccine and Serum
190 s safety, tolerability, and immunogenicity (Clinical Trials Research Institute (Chaudhary et al., 2021). Its administration
Gov, 2021). CinnaGen claimed that the Covax-19 (SpikoGen) into the body is through three doses, two injections and one nasal
vaccine was well tolerated by all patients and showed a favorable spray. The injection of the second dose of the vaccine is carried
immunogenicity profile (Zamani et al., 2011). However, no out 21 days later and after 51 days; the third dose will be inhaled
results from these studies have been published yet, and this later (Tehrantimes, 2021).
phase is ongoing. The results of its preclinical trials have not been reported yet.
In August 2021, a Phase 3 clinical study of Covax-19 Razi’s vaccine was tested in two clinical trials (Ghiasi et al., 2021).
(Spikogen) began in Iran. In this randomized, double-blind, The first phase of the study included a double-blind randomized
placebo-controlled trial, the efficacy and safety of vaccines will placebo-controlled trial initiated in Iran on 29 January 2021.

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Heidary et al. Protein Subunit Vaccines Against COVID-19

In this phase, 133 healthy volunteers, with the age range of and contains a composition of chemically synthesized peptide
18–55, were assigned to four random groups, in which the last immunogens of the S protein of the SARS-CoV-2. The vaccine
group was control. has been indicated to have ability to conjugate to a carrier protein
In the first phase of human clinical test, the vaccine proved to and adsorb on an adjuvant containing aluminum hydroxide
be safe, though mild complications were detected. Phase 2 clinical (Ryzhikov A. B. et al., 2021).
trial of the vaccine initiated in April 2021 to evaluate the safety Based on evidence, the peptides and the viral part of
and immunogenicity of the vaccine candidate. The same as phase the chimeric protein need to immunize individuals receiving
2, phase 1 was conducted in Iran and included a randomized EVCV against the SARS-CoV-2 and to activate the induction
placebo-controlled trial in which 500 volunteers aged 18–70 of protective antibodies. The producers of EVCV believe that
participated. Participants were categorized into two study groups this vaccine has stability at refrigerator temperatures for up
comprising of one vaccine group and a placebo group. The to 2 years. All people who received the vaccine developed
first group received a selected vaccine dose from phase 1, and specific antibodies against its antigen. However, the efficacy of
the second group received adjuvant only. The immunogenicity EVCV is officially uncertain and requires regulatory approval
and efficiency of the vaccine indicated in the phase 1 trial were (Abdulla et al., 2021).
confirmed in the phase 2 (Pilicheva and Boyuklieva, 2021).
SCTV01C/SCTV01E
MVC-COV1901 Vaccine The two protein subunit vaccine, called SCTV01C and
MVC-COV1901 is a recombinant protein subunit vaccine based SCTV01E, has developed by the Chinese company Sinocelltech.
on the stabilized pre-fusion SARS-CoV-2 spike protein S-2P, The vaccines were injected intra-muscular. CTV01C is a
adjuvanted with CpG 1018 (an oligodeoxynucleotide which acts Bivalent SARS-CoV-2 Trimeric Spike Protein Vaccine plus a
as a toll-like receptor 9 agonist) and aluminum hydroxide. squalene-based oil-in-water adjuvant used against SARS-CoV-2
The S-2P protein was developed by Wrap and colleagues4 at variants. 12 trials in 3 phases assessed the safety, tolerability,
the US National Institute of Allergy and Infectious Diseases, immunogenicity, and protective effect of the SCTV01C in two
and the synthetic oligodeoxynucleotide CpG 1018 adjuvant countries of China and the United Arab Emirates. In February
was developed by Dynavax Technologies (Emeryville, CA, 2022, SCTV01C entered phase 2 trials as booster shots to assess
United States). The result of a phase 1 trial showed that their ability to generate immunity in adults and adolescents
MVC-COV1901 was well tolerated and elicits robust T-cell and who have already received other vaccines. On 14 September
B-cell immune responses. The MVC-COV1901 also indicated an 2021, a phase 2/3 trial, multicenter, randomized, double-
acceptable safety profile and could elicit favorable neutralizing blinded trial (NCT05043285) assessed the safety, tolerability,
antibody titers. Besides safety, the vaccine was demonstrated to and immunogenicity of SCTV01C in 12,420 participants (18
be well tolerated. In young and older adults, it rarely causes Years and Older) in the United Arab Emirates that previously
febrile reactions. MVC-COV1901 vaccine produces high levels vaccinated with inactivated COVID-19. In April 2022, the
of neutralizing antibodies and induces anti-spike IgG titers, and company conducted phase 3 clinical trial (NCT05323461) to
its seroconversion rate is about 100% by day 57. Considering assess the safety of SCTV01C or SCTV01E on 1,800 healthy
the WHO IU and BAU conversion models, the clinical efficacy volunteers (≥18 years old) previously vaccinated with other
predicted for the vaccine is the same. Phase 2 clinical trial COVID-19 vaccine or previously diagnosed with COVID-19.
affirmed the progress of MVC-COV1901 in future phase 3 trials. SCTV01E is a COVID-19 Alpha/Beta/Delta/Omicron variants
Recently, the vaccine has received authorization for emergency S-trimer vaccine used as booster shots in phase 2. On April 4,
use in Taiwan (Hsieh et al., 2021). In an earlier investigation, another phase 3 trial for SCTV01E was conducted on 12,000
the vaccine-induced increased neutralizing antibodies with a healthy volunteers (≥12 years) formerly unvaccinated or fully
10,000-fold rise in IgG level and a mean of 50-fold higher vaccinated with Sinopharm inactivated COVID-19 vaccine or
pseudovirus neutralizing titers in each dose group than vehicle mRNA/adenovirus vectored vaccine. This vaccine is not yet
group or adjuvant control. Six days after the infection, vaccinated approved (Keech et al., 2020).
hamsters showed no weight loss and had considerably decreased
lung pathology. Moreover, the viral load levels in the lungs of
vaccinated (but not unvaccinated) animals declined to less than RECOMBINANT RBD VACCINES
detection limit. Vaccination with either 1 or 5 µg of adjuvant S-2P
induced similar immunogenicity and protection from infection ZF2001 Vaccine
(Lien et al., 2021). ZF2001 is an adjuvanted protein subunit vaccine developed by
Anhui Zhifei Longcom Biologic Pharmacy Co and the Institute of
EpiVacCorona Vaccine Medical Biology at the Chinese Academy of Medical Sciences that
EpiVacCorona Vaccine (EVCV) is an antigen-based vaccine utilizes a tandem-repeat SARS-CoV-2 spike receptor-binding
produced by the Vektor State Research Center of Virology and domain (RBD) dimer as the antigen. To better immunogenicity,
Biotechnology, Koltsovo, Russia. This vaccine has ability to the protein was designed as a tandem-repeat dimeric RBD
provoke an immune reaction against COVID-19 and is able to (Premkumar et al., 2020; Wang N. et al., 2020; Wang Q. et al.,
enhance immunity development. The EVCV is dependent on 2020). The RBD-Dimer model was created using clinical-grade
chemically synthesized peptide antigens of SARS-CoV-2 proteins Chinese hamster ovary (CHO) cell lines, and then combined with

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Heidary et al. Protein Subunit Vaccines Against COVID-19

aluminum hydroxide as an adjuvant to make the final vaccine natural infection, was investigated. Their findings revealed that
(Pourmalek et al., 2021). RBD-subunit vaccines, such as ZF2001, had much greater
Dai et al. (2020) described the structure-guided design of tolerance to 501Y.V2 than convalescents. Furthermore, a longer
a coronavirus immunogen composed of two protein subunits interval between the third and second doses of ZF2001
containing the virus spike RBD. They joined together through results in more robust 501Y.V2 neutralizing activity than the
a disulfide bond or tandem repeat. Their results demonstrated usual three-dose administration. These researchers found that
that the compared to the conventional monomeric form of SARS- launching RBD vaccines through a third-dose boost may be
CoV-2 RBD, the dimeric form of RBD increased immunogenicity suitable for fighting SARS-CoV-2 strains (Cao et al., 2021).
in a mouse model (Dai et al., 2020).
Yang et al. (2021) performed two randomized, double- Abdala (CIGB-66) Vaccine
blind, placebo-controlled, phase 1 and phase 2 trials in China. Abdala, with the technical name CIGB-66, is a Cuban
Participants in the phase 1 study were randomly allocated vaccine developed by the Center for Genetic Engineering
(2:2:1) to receive three intramuscular doses of the vaccine and Biotechnology (CIGB) and is based on the recombinant
(25 or 50 µg) or a placebo 30 days apart. In phase 2, RBD subunit of the spike protein of the SARS-CoV-2 virus.
individuals were randomized (1:1:1:1) to receive the vaccination The vaccine was produced in Pichia pastoris yeast containing
(25 or 50 µg) or placebo intramuscularly in two or three aluminum hydroxide as adjuvant. The phase 1/2 clinical trial of
doses 30 days apart. According to the findings of Phase the vaccine was initiated on December 7, 2020, and the result
1 and 2 studies, 83% of individuals developed neutralizing indicated that the vaccine does not need a carrier protein to
antibodies after two doses of the vaccine, and 97% developed acquire high levels of this indicator. The phase 3 trial began on
neutralizing antibodies after three doses. Furthermore, none of March 22, 2021 and showed 92.28% efficacy. The vaccine was
the participants had any severe adverse effects. The 25 µg/three- affirmed for emergency use on July 9. The Abdala vaccine aims
dose group had the most excellent SARS-CoV-2 neutralizing to introduce antibodies interfering with the entry of the pathogen
response (Yang et al., 2021). into cells, a fundamental mechanism create protection.
NCT04646590 is a randomized phase 3 study that is presently CIGB was selected Pichia pastoris as an expression system
ongoing. It was first registered in November 2020, with plans because of its experience in using the technological platform,
to enroll 29,000 healthy adults 18 years and older in China, which is cheaper than other platforms. As RBD is a covalently
including 750 participants aged 18–59 years and 250 participants associated glycoprotein, the saccharides of P. pastoris provide this
aged 60 years and over; 21,000 participants aged 18–59 years and domain with an immunopotentiating impact that contributes
7,000 participants aged 60 years and over will receive 25 µg of to immunogenicity. Abdala was designed by means of protein
the vaccine over a 0, 1, and 2-month schedule. According to early engineering using structural bioinformatics computational
phase 3 findings released by Zhifei, the vaccines effectiveness rate methods aimed at increasing its similarity to the SARS-CoV-2
against COVID-19 patients of any severity and delta variant was virus (Shalash et al., 2021). Recently, it has been reported that
81.76 and 77.54%, respectively. Although preliminary findings the Abdala vaccine has more than 90% effectiveness against
have not been peer-reviewed, this vaccine has been approved for severity and death, notwithstanding the prevalence of the Delta
emergency use in China and Uzbekistan since March 2021 (The variant of SARS-CoV-2 (Lemos-Perez et al., 2021). In July 2021,
Straits Times, 2021). Abdala commenced clinical trial phase I/II for children and
Huang et al. (2021) evaluated neutralization activity in 24 adolescents aged 3–18.
serum samples from subjects in two clinical trials, 12 of which
were vaccinated with BBIBP-CorV (inactivated vaccine). The BECOV2
others were immunized with ZF2001 against variant 501Y.V2 The COVID-19 vaccine candidate BECOV2, also known as
and SARS-CoV-2 wild type. Based on their results, the variant Corbevax or NEGVAC or BioE COVID-19, was developed
501Y.V2 did not escape immunity evoked by vaccines target the by Indian company Biological E. (located in Hyderabad)
whole virus (BBIBP-CorV) or S protein dimeric RBD vaccines in collaboration with a group including Baylor College of
(ZF2001) (Huang et al., 2021). Medicine [United States; Texas Children’s Hospital (Center for
Zhao et al. (2021) tested the neutralization efficacy of Vaccine Development)] and Dynavax Technologies Corporation
ZF2001 against coronavirus variants using a pseudotyped virus (United States) (Dilipkumar, 2021; Verma, 2021). This vaccine
expressing SARS-CoV-2 spike. They discovered that the ZF2001 was approved in India and Botswana but for emergency use.
vaccination maintained its neutralizing effectiveness against This vaccine is based on a recombinant protein subunit of spike
the newly discovered delta variant. However, this vaccination protein (construct of RBD N1C1 made by Baylor College of
demonstrated a more significant decrease in activity against the Medicine) in a combination of alum adjuvant with Dynavax
beta variant. Furthermore, individuals with a longer gap between Technologies Corporation’s CpG (made by Dynavax) that
the second and third doses (doses at 0, 1, and 4–6 months) had elicited a robust immune response against coronavirus (Hodgson
more significant neutralizing action (Zhao et al., 2021). et al., 2021). In phase 1/2 clinical trial (CTRI/2020/11/029032),
In a study by Cao et al. (2021), the humoral immune response the vaccine was administered intramuscularly in a two-dose
to circulating SARS-CoV-2 variants such as 501Y.V2 (B.1.351) schedule (0.5 ml; day 0 and day 28) to select a better
in plasma and neutralizing antibodies elicited by CoronaVac vaccine formulation based on overall safety and immunogenicity
(inactivated vaccine), ZF2001 (RBD-subunit vaccine), and observation. Following reported promising results in phase 1/2

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Heidary et al. Protein Subunit Vaccines Against COVID-19

clinical trials, BECOV2 has completed two phases 2/3 clinical UB-612 Vaccine
trials. After establishing the safety and immunogenicity of the The UB-612, the first COVID-19 “multi-tope” protein-peptide
vaccine in 1,268 COVID-19-negative adult subjects (18–80 years) vaccine, was produced by United Biomedical Inc. Asia in Taipei.
at two doses with 28-day intervals in the first phase 2/3 clinical The vaccine comprises of eight components and includes a
trial (CTRI/2021/06/034014), the vaccine is further investigated strong S1-RBD component connected to a single-chain fragment
in second phase 2/3 clinical trial (CTRI/2021/10/037066) on region (sFC) of a human IgG1. This region, which functions
624 children and adolescents aged between < 18 years and as the principal neutralizing domain of the virus, facilitates cell
≥ 5 years with a booster dose on Day 208. In this phase, the attachment. The UB-612 vaccine consists of a proprietary peptide
vaccine induces a robust immune response without any severe UBITh1a , five peptides with Th or CTL epitopes, and aluminum
R

AEs among the pediatric population as young as 5 years old. phosphate adjuvant. With the aim of inducing a broad immune
Currently, the vaccine is approved for children between 5 and response, the vaccine was designed and was able to mitigate
12 but is restricted to use in emergency situations. In phase viral load and block COVID-19 infection in mice and monkeys.
3 (CTRI/2021/08/036074), the BECOV2 vaccine was compared In rats, UB-612 toxicity has been displayed to have appropriate
with Covishield on more than 2,140 COVID-19-negative adult safety (Chaudhary et al., 2021; Hasanzadeh et al., 2021).
subjects (18–80 years) at 33 study sites across India. This study In Guirakhoo et al.’s (2020) study, UB-612 vaccine was
demonstrates that the BECOV2 vaccine induces a superior composed of eight components, six synthetic peptides, a
immune response than the Covishield vaccine. Also, it had 50% proprietary CpG TLR-9 agonist, and aluminum phosphate
fewer adverse events than Covishield. A researcher showed that adjuvant. The phase 1 trial and additional trials conducted
this vaccine’s effectiveness is 90% against the Ancestral-Wuhan globally have denoted that the vaccine is highly promising and a
strain and 80% against the Delta strain to prevent symptomatic differentiated candidate for inhibiting SARS-CoV-2 transmission
infections (Speiser and Bachmann, 2020). and COVID-19 disease.
According to the pre-clinical investigations in guinea pigs,
FINLAY-FR-2 Vaccine rats, and mice, the UB-612 vaccine could induce extremely high
The Finlay Vaccine Institute in Cuba developed the FINLAY-FR-2 titers of neutralizing antibodies with S1-RBD:hACE2 inhibition
(Soberana 02) COVID-19 vaccine. It is a conjugate vaccine, which activities and could generate balanced Th1/Th2 response toward
means that the viral antigen, the RBD, is chemically coupled the Th1 polarity. The vaccine also could induce balanced B cell
to the tetanus toxoid to keep it stable. Aluminum hydroxide is activation and enhance T cell immune response in human body,
used as an adjuvant in Soberana 2 to strengthen the immune hence providing exceptional protection. Moreover, the UB-612
(Valdes-Balbin et al., 2021). vaccine could generate a favorable immune response in humans
Valdes-Balbin et al. (2021) demonstrated in pre-clinical and is well tolerated without serious AVs. The vaccine does not
investigations that macromolecular constructs containing need to be transported and stored in an ultra-low temperature.
recombinant RBD coupled to tetanus toxoid elicit a robust This feature is speculated to be more competitive than other
immunological response in laboratory animals. RNA vaccines produced by companies such as Pfizer/BNT
According to the Cuban Public Registry of Clinical Trials, a (Kanno et al., 2021).
phase 1 clinical trial with 40 volunteers began in October 2020,
with an open, sequential, and adaptive study to evaluate the
vaccine’s safety, reactogenicity, and immunogenicity (Soberana ReCOV
02, 2021). China’s Jiangsu Rec-Biotechnology developed the ReCOV
Following early findings, the vaccine based on RBD6-TT/alum vaccine. This recombinant protein two-component COVID-19
was advanced to a phase 2 clinical study in December 2020. vaccine is grown in Chinese hamster ovary cells. This vaccine
Phase 2a of the vaccination included 100 Cubans, while Phase 2b consists of the RBD of the spike protein (protein engineering
included 810 volunteers aged 19–80 years old (Universo Online, platforms) and a novel adjuvant, which enters the host cells by
2021). Initial studies indicated that the vaccination elicited an binding to angiotensin-converting enzyme 2. After vaccination,
immunological response after 14 days (OnCubaNews English, the host will induce neutralizing antibodies against the spike
2020). Phase 3 commenced at the beginning of March as initially protein to protect them from infection with the native virus
scheduled. The 44,010 volunteers were divided into three groups: (Biorender, 2021a). On March 26, a phase 1 clinical trial was
some received two doses of the vaccine 28 days apart; another launched on 160 healthy volunteers from 18 to 80 years, at two
group will get two doses plus a third immune booster (Soberana doses with 21 days interval, to evaluate the safety, reactogenicity,
Plus), and the third a placebo (ABS CBN, 2021). and immunogenicity in New Zealand. Following vaccine effective
Pasteur Institute of Iran performed Phase 3 on 24,000 people at generating immunity, this company incorporated Shenzhen
aged 18–80 years old in 8 cities as part of the collaboration with Rhegen Biotechnology and founded the company of Wuhan
other nations to develop the COVID-19 vaccine (IRCT, 2021). Rhecogen Biotechnology on September 29. In late 2021, this new
Although clinical study results have not yet been published, company conducted a Phase 2/3 trial to evaluate the efficacy,
preliminary reports suggest a 62% efficacy after just two doses safety, and immunogenicity of ReCOV in 18–80 years adult
(Acosta, 2021). When used in conjunction with a Plus booster volunteers in the Philippines. The data results showed that
dose, the vaccination was also 91.2% effective, according to ReCOV induced high levels of neutralizing antibodies against
BioCubaFarma (The Indian Express, 2021). variants of coronavirus without side effects. In April 2022, it

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Heidary et al. Protein Subunit Vaccines Against COVID-19

received approval to extend the trial to the U.A.E. ReCOV has and transient. The occurrence of significant adverse events (AVs)
not yet been approved (Ghiasi et al., 2021). was minimal and equivalent in both groups (Heath et al., 2021).
PREVENT-19 trial NCT04611802 is a randomized phase 3
study, including 29,960 adult volunteers that performed in the
United States and Mexico (Shirmohammadi-Khorram et al.,
NANOPARTICLE VACCINES 2019). The participants were entirely (100%) immune to both
mild and severe diseases. The vaccine was 90.0% effective in
NVX-CoV2373 Vaccine avoiding symptomatic COVID-19 illness in individuals at high
The NVX-CoV2373 protein subunit vaccine developed by risk of acquiring COVID-19 issues. According to safety data, the
Novavax contains Matrix-M1 adjuvant2 and a recombinant experimental vaccine was usually well-tolerated (Johnson, 2021).
SARS-CoV-2 (rSARS-CoV-2) nanoparticle vaccine 3 from the
full-length, wild-type SARS-CoV-2 spike glycoprotein. It targets GBP510 Vaccine
antibody and vaccine development because it enhances viral The GBP510 (recently known as SKYCovione) is a self-
attachment to the host cell’s human angiotensin-converting assembled nanoparticle vaccine developed by the South Korean
enzyme 2 (hACE2) receptor. Using the saponin-based matrix company SK Bioscience and the Institute for Protein Design
M1 adjuvant, the lack of cell-mediated immune responses, which (IPD) at the University of Washington with a combination of
characterize protein subunit vaccinations, was overcome (Du GlaxoSmithKline’s (GSK; a British multinational pharmaceutical
et al., 2009; Bengtsson et al., 2016; Keech et al., 2020; Tai et al., company) adjuvant (AS03). GBP510 is a recombinant protein-
2020). Matrix M-adjuvanted NVX-CoV2373 was evaluated for based vaccine that targets the RBD of the SARS-CoV-2 spike
immunogenicity in animal models such as rats and baboons. protein (Smith et al., 2020).
In pre-clinical studies, NVX-CoV2373 generated antibodies that The phase I/II results demonstrate a high level of neutralizing
blocked the binding of spike protein to cellular receptors and antibody with a 100% seroconversion rate in healthy adults (aged
protected against infection (Mandolesi et al., 2020; Tian et al., 19–85 years) given the adjuvanted vaccine. The phase III trial
2021). (NCT05007951) of the vaccine was started with 4,037 participants
Following early encouraging findings in animal models, (570 participants in Korea and 3,467 other countries) over
Novavax launched a Phase 1/2 trial. Clinical results from 18-year-old in Thailand, Vietnam, New Zealand, Ukraine, the
Novavax’s phase 1 and 2 trials revealed that a two-dose regimen Philippines, and South Korea in cooperation with 16 institutions.
given 21 days apart was safe and generated more significant The results show this vaccine only elicited a superior neutralizing
levels of coronavirus antibodies than those observed in COVID- antibody (over 95%) than AstraZeneca’s Covid-19 vaccine (79%,
19 survivors. The vaccination also increased T cells, another control vaccine), Vaxzevria, in subjects aged > 65 but induced
component of the human immune system (Keech et al., 2020). 2.93 times notarization antibody titer 2 weeks after of the second
Novavax developed a Randomized, Observer-Blinded, dose that of a control vaccine. Also, this vaccine showed a
Placebo-Controlled Study phase 2 a/b study in South Africa to favorable safety profile compared to the control vaccine (mild or
evaluate the effectiveness, safety, and immunogenicity of the moderate adverse reactions) in this vaccine. This vaccine is not
Novavax vaccine in adults subjects living without HIV and adults yet approved (PMLiVE, 2021).
living with HIV (ClinicalTrials.gov NCT04533399). At the time
of this Phase, B.1.351 variant was operating throughout the
country and beyond. An early analysis of this clinical trial data CONCLUSION AND OUTLOOK
revealed that total vaccination effectiveness was 49%. Among
HIV-negative subjects, Novavax was 60% effective. Furthermore, Producing effective vaccines based on Protein Subunit against
vaccine effectiveness against the B.1.351 variant was 51.0% SARS-CoV-2 Infection as rather new technology has become a
(Shinde et al., 2021). Severe and medically attended adverse use full choice. Here, we reviewed 16 vaccines passing phases
effects were uncommon. Consequently, the findings of this 3 and 4 from different countries, Australia, the United States,
clinical trial demonstrated that the NVX-CoV2373 vaccination India, Russia, China, South Korea, and Iran. SARS-CoV-2 spike
had greater effectiveness in preventing Covid-19 among protein S is a large, trimeric glycoprotein that plays the most
HIV-negative individuals (Callaway and Mallapaty, 2021). important roles in viral attachment, fusion and entry, and serves
A phase 3 randomized, observer-blinded, placebo-controlled as a target for the development of antibodies, entry inhibitors
study was performed at 33 locations in the United Kingdom and vaccines. In these vaccines, different protein particles
between September and November 2020, recruiting up to 9,000 such as full-length wild-type SARS-CoV-2 spike glycoprotein,
individuals to evaluate the Novavax’s effectiveness, safety, and SARS-CoV-2 spike protein (D614), tandem-repeat SARS-CoV-2
immunogenicity (NCT04583995). The participants’ ages varied spike receptor-binding domain (RBD) dimer. The utilization of
from 18 to 84. Two doses of Novavax vaccine given 21 days apart aluminum hydroxide as adjuvant was common in most vaccines.
in adults offered 89.7% protection against symptomatic Covid-19 In addition, CpG 1018 in MVC-COV1901 and Matrix-M1 in
caused by both B.1.1.7 and non-B.1.1.7 variants, according to NVX-CoV2373 were some other adjuvants.
the results (overall efficacy). This vaccination was 86.3% effective Overall, the occurrence of adverse events in all reviewed
against B.1.1.7 (or alpha) variants and 96.4% effective against Protein Subunit Vaccines was uncommon or minimal. In
non-B.1.1.7 variants. In most instances, reactogenicity was minor Iran COVAX-19, Razi Cov Pars and Soberana-02 are three

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Heidary et al. Protein Subunit Vaccines Against COVID-19

Protein Subunit Vaccines that are approved for use. From an to distribute. Novavax is shown to be highly effective in
administration way point of view, except Razi Cov Pars that clinical trials. Ninety percent effective against lab-confirmed,
uses two injections and one nasal spray dose, other vaccines symptomatic infection and 100% against moderate and severe
have two or three intramuscular doses. Vaccines with intranasal disease in Phase 3 trial results released. In a Phase 3 study
spray formulation can be easily administered and would be conducted in the United States and Mexico during a period
beneficial as mucosal immunity is known as the first line of in which multiple variants (Alpha, Beta, and Delta) were in
defense against the virus. Early protection against COVID-19 circulation, vaccine efficacy against mild, moderate, or severe
infection is related to the secretion of mucosal IgA in the upper COVID-19 was 90%.
respiratory tract during initial contact with the SARS-CoV- Based on clinical trial conducted in the United Kingdom, the
2 virus. In conclusion, despite all efforts to produce effective efficacy was 86.3% against the B.1.1.7 (alpha) variant and 96.4%
vaccines, the appearance of new mutated SARS-CoV-2 strains is a against non-B.1.1.7 variants. On the other hand, based on clinical
serious challenge. Present vaccine proficiency must be evaluated trial performed in the United States and Mexico, efficacy against
and their efficiency reported. the alpha variant was 93.6% and against any variant was 92.6%.
Based on available documentation from clinical trials, five Moreover, the efficacy against the latter variants was reported
of the protein subunit vaccines have reported the efficacy 100%. Novavax says its vaccine can generate an immune response
which is as follows: Abdala (92.28%), SOBERANA 02 Plus against Omicron, but scientists are still learning about this.
(92.4%), Novavax (92.6%), EPIVACCORONA (79%), and SCB- Prevention of severe disease and mortality is the main objective
2019 (67.2%). Among these, Abdala, SOBERANA 02 Plus, and of vaccination. Hence, the occurrence of serious adverse effect
Novavax have shown high efficacy and safety profile, but more was low and transient, and also no hospitalizations or death was
precise, the results of Phase III trials of Novavax, SOBERANA 02 occurred among the vaccinated recipients.
Plus, and SCB-2019 have published by more detailed analyses.
In addition to favorable results of these candidated vaccines,
in our view, Novavax may be one of the promising vaccines AUTHOR CONTRIBUTIONS
because it could elicits protective immunity against broad
spectrum of variants. MH and SK contributed to the conception, design of the work,
This vaccine contains the spike protein of the coronavirus and acquisition. VK, MS, RG, MM, MS, HG, MH, and SK
itself, but formulated as a nanoparticle, which cannot cause contributed in drafting of the manuscript, revising and final
disease. It is simpler to make than some of the other vaccines approval of the version to be published. All authors contributed
and can be stored in a refrigerator (2–8◦ C), making it easier to the article and approved the submitted version.

REFERENCES Bravo, L., Smolenov, I., Han, H. H., Li, P., Hosain, R., Rockhold, F., et al. (2022).
Efficacy of the adjuvanted subunit protein COVID-19 vaccine, SCB-2019: a
Abdulla, Z. A., Al-Bashir, S. M., Al-Salih, N. S., Aldamen, A. A., and phase 2 and 3 multicentre, double-blind, randomised, placebo-controlled trial.
Abdulazeez, M. Z. (2021). A Summary of the SARS-CoV-2 Vaccines and Lancet 399, 461–472. doi: 10.1016/S0140-6736(22)00055-1
Technologies Available or under Development. Pathogens 10:788. doi: 10.3390/ Callaway, E., and Mallapaty, S. (2021). Novavax covid vaccine protects people
pathogens10070788 against variants. Nature 590:17. doi: 10.1038/d41586-021-00268-9
ABS CBN (2021). Cuban-Developed Vaccine Enters Phase III Trial. Philippines: Cao, Y., Yisimayi, A., Bai, Y., Huang, W., Li, X., Zhang, Z., et al. (2021). Humoral
ABS CBN. immune response to circulating SARS-CoV-2 variants elicited by inactivated
Acosta, N. (2021). Cuba Encouraged by Early Efficacy Results of COVID-19 Vaccine. and RBD-subunit vaccines. Cell Res. 31, 732–741. doi: 10.1038/s41422-021-
London: Reuters. 00514-9
Aguilar-Guerra, T. L., Fajardo-Díaz, E. M., and Gorry, C. (2021). Cuba’s National Chaudhary, S., El-Shorbagi, A. N., Gupta, R. K., and Kumar, A. (2021). The
Regulatory Authority & COVID-19: olga Lidia Jacobo-Casanueva MS Director. Recent Updates on Approaches and Clinical Trials Status of Covid-19 Vaccines
Center for State Control of Medicines and Medical Devices (CECMED). Developed Globally. Biomed. Pharmacol. J. 14, 1109–1124. doi: 10.13005/bpj/
MEDICC Rev. 23, 9–14. doi: 10.37757/MR2021.V23.N3.3 2214
Arashkia, A., Jalilvand, S., Mohajel, N., Afchangi, A., Azadmanesh, K., Salehi- Chavda, V. P., Vora, L. K., and Vihol, D. R. (2021). COVAX-19 R Vaccine:
Vaziri, M., et al. (2021). Severe acute respiratory syndrome-coronavirus-2 spike completely blocks virus transmission to non-immune individuals. Clin. Compl.
(S) protein based vaccine candidates: state of the art and future prospects. Rev. Med. Pharmacol. 1:100004. doi: 10.1016/j.ccmp.2021.100004
Med. Virol. 31:e2183. doi: 10.1002/rmv.2183 Cinnagen (2021). A Phase III, Randomized, Two-armed, Double-blind, Placebo
Baird, F. J., and Lopata, A. L. (2014). The dichotomy of pathogens and allergens in controlled trial to evaluate efficacy and safety of an adjuvanted recombinant
vaccination approaches. Front. Microbiol. 5:365. doi: 10.3389/fmicb.2014.00365 SARS-CoV-2 spike (S) protein subunit vaccine (SpikoGen R ) produced by
Bengtsson, K. L., Song, H., Stertman, L., Liu, Y., Flyer, D. C., Massare, M. J., et al. CinnaGen Co. Tehran: Cinnagen.
(2016). Matrix-M adjuvant enhances antibody, cellular and protective immune Clinical Trials Gov (2021). A Phase II, Randomized, Two-armed, Double-
responses of a Zaire Ebola/Makona virus glycoprotein (GP) nanoparticle blind, Placebo-controlled Trial to Evaluate the Safety, Tolerability, and
vaccine in mice. Vaccine 34, 1927–1935. doi: 10.1016/j.vaccine.2016.02.033 Immunogenicity of an Adjuvanted Recombinant SARS-CoV-2 Spike (S)
Bill, R. M. (2015). Recombinant protein subunit vaccine synthesis in microbes: a Protein Subunit Vaccine Candidate (SpikoGen). Available online at: https://
role for yeast?. J. Pharm. Pharmacol. 67, 319–328. doi: 10.1111/jphp.12353 clinicaltrials.gov/show/NCT04944368
Biorender (2021a). ReCOV. Available online at: https://biorender.com/covid- Clinicaltrials Gov (2022). Safety, Reactogenicity and Immunogenicity Study of
vaccine-tracker/details/v-CV64/-67 (accessed on Mar 26, 2021). ReCOV. Available online at: https://clinicaltrials.gov/ct2/show/NCT04818801?
Biorender (2021b). Unnamed Inactive Vaccine - Jiangsu. Available online at: cond=Recov&draw=2&rank=1 (accessed on May 5, 2021).
https://biorender.com/covid-vaccine-tracker/details/v-CV52/inactivated- Clover Biopharmaceuticals (2021a). Clover Biopharmaceuticals Completes
covid-19-vaccine-54 (accessed Feb 17, 2021). Enrollment of Adult and Elderly Population in SPECTRA Global Phase

Frontiers in Microbiology | www.frontiersin.org 11 July 2022 | Volume 13 | Article 927306

Heidary et al. Protein Subunit Vaccines Against COVID-19

2/3 Clinical Trial for its COVID-19 Vaccine Candidate. Chengdu: Clover Hsieh, S. M., Liu, M. C., Chen, Y. H., Lee, W. S., Hwang, S. J., Cheng, S. H., et al.
Biopharmaceuticals. (2021). Safety and immunogenicity of CpG 1018 and aluminium hydroxide-
Clover Biopharmaceuticals (2021b). Clover’s COVID-19 Vaccine Candidate adjuvanted SARS-CoV-2 S-2P protein vaccine MVC-COV1901: interim results
Demonstrates 79% Efficacy Against Delta in Global Phase 2/3 SPECTRA of a large-scale, double-blind, randomised, placebo-controlled phase 2 trial in
Trial Dominated by Variants of Concern and Interest. Chengdu: Clover Taiwan. Lancet Resp. Med. 9, 1396–1406. doi: 10.1016/S2213-2600(21)00402-1
Biopharmaceuticals. Huang, B., Dai, L., Wang, H., Hu, Z., Yang, X., Tan, W., et al. (2021). Serum sample
Currier, A. W., Jeshurin, M. C., and Sampson, V. B. (2021). SARS-CoV-2 Targets neutralisation of BBIBP-CorV and ZF2001 vaccines to SARS-CoV-2 501Y. V2.
and COVID-19 Vaccines. COVID 1, 608–621. doi: 10.3390/covid1030051 Lancet Microbe 2:e285. doi: 10.1016/S2666-5247(21)00082-3
Dai, L., Zheng, T., Xu, K., Han, Y., Xu, L., Huang, E., et al. (2020). A universal IRCT. (2021). Efficacy, Safety, and Immunogenicity of Soberana Recombinant
design of betacoronavirus vaccines against COVID-19, MERS, and SARS. Cell Vaccine (Product of Finlay Institute) based on RBD Protein Subunit of Sars-
182, 722–733.e11. doi: 10.1016/j.cell.2020.06.035 Cov-2 in a 2-Dose Regimen with and without a Booster Dose: a Double-
Deeks, J. J., Dinnes, J., Takwoingi, Y., Davenport, C., Spijker, R., Taylor-Phillips, Blind, Randomized, Placebo-Controlled Phase III Clinical Trial in the Iranian
S., et al. (2020). Antibody tests for identification of current and past infection population of 18-80 Years. New Delhi: IRCT.
with SARS-CoV-2. Cochrane Database Syst. Rev. 6:CD013652. doi: 10.1002/ Jacob, S. T., Crozier, I., Fischer, W. A., Hewlett, A., Kraft, C. S., de La Vega,
14651858.CD013652 M.-A., et al. (2020). Ebola virus disease. Nat. Rev. Dis. Primers 6, 1–31. doi:
Dilipkumar. (2021). Info about Corbevax/Becov2 Vaccine developed by Biological E, 10.1038/s41572-020-0147-3
BCM and Dynavax. Available online at: https://dilipkumar.in/articles/health/ Johnson, L. A. (2021). Large Study Finds that Vaccine Maker Novavax’s Shot is About
info-about-cobrevax-becov2-vaccine-being-developed-by-biological-e.html 90% Effective. New York: The Associated Press.
(accessed on 10 Jun, 2021). Kandimalla, R., Chakraborty, P., Vallamkondu, J., Chaudhary, A., Samanta, S.,
Dong, Y., Dai, T., Wei, Y., Zhang, L., Zheng, M., and Zhou, F. A. (2020). Systematic Reddy, P. H., et al. (2021). Counting on COVID-19 vaccine: insights into the
review of SARS-CoV-2 vaccine candidates. Signal Transduct. Target. Ther. current strategies, progress and future challenges. Biomedicines 9:1740. doi:
5:237. doi: 10.1038/s41392-020-00352-y 10.3390/biomedicines9111740
Du, L., He, Y., Zhou, Y., Liu, S., Zheng, B. J., and Jiang, S. (2009). The spike protein Kanno, A. I., Barbosa, M. M., Moraes, L., and Leite, L. C. (2021). SARS-CoV-
of SARS-CoV—a target for vaccine and therapeutic development. Nat. Rev. 2 vaccine development and how Brazil is contributing. Genet. Mol. Biol.
Microbiol. 7, 226–236. doi: 10.1038/nrmicro2090 44:e20200320. doi: 10.1590/1678-4685-GMB-2020-0320
Dunkle, L. M., Kotloff, K. L., Gay, C. L., Áñez, G., Adelglass, J. M., Barrat Keech, C., Albert, G., Cho, I., Robertson, A., Reed, P., Neal, S., et al. (2020). Phase
Hernández, A. Q., et al. (2021). Efficacy and Safety of NVX-CoV2373 in Adults 1–2 trial of a SARS-CoV-2 recombinant spike protein nanoparticle vaccine. New
in the United States and Mexico. New Engl. J. Med. 386, 531–543. doi: 10.1056/ Engl. J. Med. 383, 2320–2332. doi: 10.1056/NEJMoa2026920
NEJMoa2116185 Khademi, F., Yousefi-Avarvand, A., Sahebkar, A., Ghanbari, F., and Vaez, H. (2018).
Eroglu, B., Nuwarda, R. F., Ramzan, I., and Kayser, V. A. (2021). Narrative Review Bacterial Co-infections in HIV/AIDS-positive Subjects: a Systematic Review
of COVID-19 Vaccines. Vaccines 10:62. doi: 10.3390/vaccines10010062 and Meta-analysis. Folia Med. 60, 339–350. doi: 10.2478/folmed-2018-0007
Garçon, N., Vaughn, D. W., and Didierlaurent, A. M. (2012). Development and Khan, Z., Ghafoor, D., Khan, A., Ualiyeva, D., Khan, S. A., Bilal, H., et al. (2020).
evaluation of AS03, an Adjuvant System containing α-tocopherol and squalene Diagnostic approaches and potential therapeutic options for coronavirus
in an oil-in-water emulsion. Exp. Rev. Vaccines 11, 349–366. doi: 10.1586/erv. disease (COVID-19). New Microbes New Infect. 38:100770. doi: 10.1016/j.nmni.
11.192 2020.100770
Ghiasi, N., Arabsorkhi, M., Hoseyni, T., Esfandiari, K., Sadighpour, T., and Khoshnood, S., Arshadi, M., Akrami, S., Koupaei, M., Ghahramanpour, H.,
Jahantigh, H. (2021). Efficacy and side effects of Sputnik V, Sinopharm Shariati, A., et al. (2022). An overview on inactivated and live-attenuated
and AstraZeneca vaccines to stop COVID-19; a review and discussion. SARS-CoV-2 vaccines. J. Clin. Lab. Anal. 36:e24418. doi: 10.1002/jcla.24418
Immunopathol. Persa 7:31. doi: 10.34172/ipp.2021.31 Koupaei, M., Mohamadi, M. H., Yashmi, I., Shahabi, A. H., Shabani, A. H., Heidary,
Goepfert, P. A., Fu, B., Chabanon, A.-L., Bonaparte, M. I., Davis, M. G., Essink, M., et al. (2022a). Clinical manifestations, treatment options, and comorbidities
B. J., et al. (2021). Safety and immunogenicity of SARS-CoV-2 recombinant in COVID-19 relapse patients: a systematic review. J. Clin. Lab. Anal. 36:e24402.
protein vaccine formulations in healthy adults: interim results of a randomised, doi: 10.1002/jcla.24402
placebo-controlled, phase 1–2, dose-ranging study. Lancet Infect. Dis. 21, 1257– Koupaei, M., Shadab Mehr, N., Mohamadi, M. H., Asadi, A., Abbasimoghaddam,
1270. doi: 10.1016/S1473-3099(21)00147-X S., Shekartabar, A., et al. (2022b). Clinical symptoms, diagnosis, treatment,
Guirakhoo, F., Kuo, L., Peng, J., Huang, J. H., Kuo, B., Lin, F., et al. (2020). A novel and outcome of COVID-19-associated encephalitis: a systematic review of case
sars-cov-2 multitope protein/peptide vaccine candidate is highly immunogenic reports and case series. J. Clin. Lab. Anal 36:e24426. doi: 10.1002/jcla.24426
and prevents lung infection in an adeno associated virus human angiotensin- Koupaei, M., Naimi, A., Moafi, N., Mohammadi, P., Tabatabaei, F. S., Ghazizadeh,
converting enzyme 2 (aav hace2) mouse model. bioRxiv [Preprint]. doi: 10. S., et al. (2021). Clinical Characteristics. Diagnosis, Treatment, and Mortality
1101/2020.11.30.399154 Rate of TB/COVID-19 Coinfectetd Patients: a Systematic Review. Front. Med.
Hasanzadeh, A., Alamdaran, M., Ahmadi, S., Nourizadeh, H., Bagherzadeh, 8:740593. doi: 10.3389/fmed.2021.740593
M. A., Jahromi, M. A. M., et al. (2021). Nanotechnology against COVID- Kumar, A., and Kumar, A. (2021). Mucosal and transdermal vaccine delivery
19: immunization, diagnostic and therapeutic studies. J. Control. Release 336, strategies against COVID-19. Drug Deliv. Transl. Res. 12, 968–972. doi: 10.1007/
354–374. doi: 10.1016/j.jconrel.2021.06.036 s13346-021-01001-9
Heath, P. T., Galiza, E. P., Baxter, D. N., Boffito, M., Browne, D., Burns, F., et al. Lemos-Perez, G., Chavez-Valdes, S., Gonzalez-Formental, H., Freyre-Corrales, G.,
(2021). Safety and efficacy of NVX-CoV2373 Covid-19 vaccine. New Engl. J. Vazquez-Arteaga, A., Alvarez-Acevedo, B., et al. (2021). Elevated antibody titers
Med. 385, 1172–1183. doi: 10.1056/NEJMoa2107659 in Abdala vaccinees evaluated by Elecsys R anti-SARS-CoV-2 S highly correlate
Hernandez-Bernal, F., del Carmen Ricardo-Cobas, M., Martin-Bauta, Y., Navarro- with UMELISA SARS-CoV-2 ANTI RBD, ACE-2 binding inhibition and viral
Rodriguez, Z., Piñera-Martínez, M., Quintana-Guerra, J., et al. (2021). Safety, neutralization assays. medRxiv [Preprint]. doi: 10.1101/2021.10.18.21265169
tolerability, and immunogenicity of a SARS-CoV-2 recombinant spike protein Li, L., Honda-Okubo, Y., Huang, Y., Jang, H., Carlock, M. A., Baldwin, J., et al.
vaccine: a randomised, double-blind, placebo-controlled, phase 1-2 clinical trial (2021). Immunisation of ferrets and mice with recombinant SARS-CoV-2
(ABDALA Study). medRxiv [Preprint]. doi: 10.1101/2021.11.30.21267047 spike protein formulated with Advax-SM adjuvant protects against COVID-19
Hodgson, S. H., Mansatta, K., Mallett, G., Harris, V., Emary, K. R., and Pollard, infection. Vaccine 39, 5940–5953. doi: 10.1016/j.vaccine.2021.07.087
A. J. (2021). What defines an efficacious COVID-19 vaccine? A review of the Liang, J. G., Su, D., Song, T.-Z., Zeng, Y., Huang, W., Wu, J., et al. (2021). S-
challenges assessing the clinical efficacy of vaccines against SARS-CoV-2. Lancet Trimer, a COVID-19 subunit vaccine candidate, induces protective immunity
Infect. Dis. 21, e26–e35. doi: 10.1016/S1473-3099(20)30773-8 in nonhuman primates. Nat. Commun. 12, 1–12. doi: 10.1038/s41467-021-
Hosier, H., Farhadian, S. F., Morotti, R. A., Deshmukh, U., Lu-Culligan, A., 21634-1
Campbell, K. H., et al. (2020). SARS–CoV-2 infection of the placenta. J. Clin. Lien, C. E., Lin, Y. J., Chen, C., Lian, W. C., Kuo, T. Y., Campbell, J. D., et al.
Investig. 130, 4947–4953. doi: 10.1172/JCI139569 (2021). CpG-adjuvanted stable prefusion SARS-CoV-2 spike protein protected

Frontiers in Microbiology | www.frontiersin.org 12 July 2022 | Volume 13 | Article 927306

Heidary et al. Protein Subunit Vaccines Against COVID-19

hamsters from SARS-CoV-2 challenge. Sci. Rep. 11:8761. doi: 10.1038/s41598- the prevention of Covid-19, in volunteers aged 18-60 years (phase I-Ii). Russ. J.
021-88283-8 Infect. Immun. 11, 283–296. doi: 10.15789/2220-7619-ASB-1699
Liu, H., Su, D., Zhang, J., Ge, S., Li, Y., Wang, F., et al. (2017). Improvement of Sanofi (2021b). Sanofi and Gsk COVID-19 Vaccine Candidate Demonstrates Strong
pharmacokinetic profile of TRAIL via trimer-tag enhances its antitumor activity Immune Responses Across all Adult Age Groups in Phase 2 Trial. Paris: Sanofi.
in vivo. Sci. Rep. 7, 1–11. doi: 10.1038/s41598-017-09518-1 Sanofi (2021a). Phase 3 Clinical Trial for COVID-19 Recombinant Protein Vaccine
Mahdizade Ari, M., Mohamadi, M. H., Shadab Mehr, N., Abbasimoghaddam, Candidate. Paris: Sanofi.
S., Shekartabar, A., Heidary, M., et al. (2022). Neurological manifestations in Sanofi (2021c). Sanofi and GSK Initiate Global Phase 3 Clinical Efficacy Study of
patients with COVID-19: a systematic review and meta-analysis. J. Clin. Lab. COVID-19 Vaccine Candidate. Paris: Sanofi.
Anal. 36:e24403. doi: 10.1002/jcla.24403 Sanofi (2021d). Study of Adjuvanted Recombinant COVID-19 Vaccine Candidate as
Mandolesi, M., Sheward, D. J., Hanke, L., Ma, J., Pushparaj, P., Vidakovics, a Primary Series and as a Booster Dose Against COVID. Paris: Sanofi.
L. P., et al. (2020). SARS-CoV-2 protein subunit vaccination elicits potent Shalash, A. O., Hussein, W. M., Skwarczynski, M., and Toth, I. (2021). Key
neutralizing antibody responses. BioRxiv [Preprint]. doi: 10.1101/2020.07.31. Considerations for the Development of Safe and Effective SARS-CoV-2 Subunit
228486 Vaccine: a Peptide-Based Vaccine Alternative. Adv. Sci. 8:2100985. doi: 10.1002/
Martínez-Flores, D., Zepeda-Cervantes, J., Cruz-Reséndiz, A., Aguirre-Sampieri, advs.202100985
S., Sampieri, A., and Vaca, L. S. A. R. S. (2021). CoV-2 Vaccines Based on the Shinde, V., Bhikha, S., Hoosain, Z., Archary, M., Bhorat, Q., Fairlie, L., et al. (2021).
Spike Glycoprotein and Implications of New Viral Variants. Front. Immunol. Efficacy of NVX-CoV2373 Covid-19 Vaccine against the B. 1.351 Variant. New
12:701501. doi: 10.3389/fimmu.2021.701501 Engl. J. Med. 384, 1899–1909. doi: 10.1056/NEJMoa2103055
Naimi, A., Yashmi, I., Jebeleh, R., Imani Mofrad, M., Azimian Abhar, S., Jannesar, Shirmohammadi-Khorram, N., Tapak, L., Hamidi, O., and Maryanaji, Z. (2019). A
Y., et al. (2022). Comorbidities and mortality rate in COVID-19 patients with comparison of three data mining time series models in prediction of monthly
hematological malignancies: a systematic review and meta-analysis. J. Clin. Lab. brucellosis surveillance data. Zoonoses Public Health 66, 759–772. doi: 10.1111/
Anal. 36:e24387. zph.12622
OnCubaNews English (2020). Cuba negotiates with other countries to Smith, T. R., Patel, A., Ramos, S., Elwood, D., Zhu, X., Yan, J., et al. (2020).
develop phase 3 of Soberana 02 vaccine. Available online at: https: Immunogenicity of a DNA vaccine candidate for COVID-19. Nat. Commun.
//oncubanews.com/en/cuba/cuba-negotiates-with-other-countries-to- 11, 1–13.
develop-phase-3-of-soberana-02-vaccine/ (accessed on Jan 24, 2021). Soberana 02. (2021). Registro Público Cubano de Ensayos Clínicos”. Cuban
Pasteur, S. (2020). Study of Recombinant Protein Vaccine Formulations Against Registry of Clinical Trials (in Spanish)
COVID-19 in Healthy Adults 18 Years of Age and Older. Geneva: WHO. Speiser, D. E., and Bachmann, M. F. (2020). COVID 19: mechanisms of vaccination
Pasteur, S. (2021). Study of Recombinant Protein Vaccine With Adjuvant Against and immunity. Vaccines 8:404. doi: 10.3390/vaccines8030404
COVID-19 in Adults 18 Years of Age and Older. Geneva: WHO. Tai, W., He, L., Zhang, X., Pu, J., Voronin, D., Jiang, S., et al. (2020).
Peeridogaheh, H., Golmohammadi, M. G., and Pourfarzi, F. (2013). Evaluation Characterization of the receptor-binding domain (RBD) of 2019 novel
of ELISA and Brucellacapt tests for diagnosis of human Brucellosis. Iran. J. coronavirus: implication for development of RBD protein as a viral attachment
Microbiol. 5, 14–18. inhibitor and vaccine. Cell. Mol. Immunol. 17, 613–620. doi: 10.1038/s41423-
Philippidis, A. (2021). Six Up-and-Coming COVID-19 Vaccines: besides 020-0400-4
Pfizer/BioNTech, Moderna, and J&J, many other biopharmas have jabs in the Tehrantimes (2021). COVID-19 Vaccine Razi Cov Pars Starts Third Clinical
clinic that have shown significant protection against SARS-CoV-2. Genet. Eng. Trial Phase. Available online at: https://www.tehrantimes.com/news/464365/
Biotechnol. News 41, 24–68. COVID-19-vaccine-Razi-Cov-Pars-starts-third-clinical-trial-phase (accessed
Pilicheva, B., and Boyuklieva, R. (2021). Can the Nasal Cavity Help Tackle COVID- on Aug 27, 2021).
19? Pharmaceutics 13:1612. doi: 10.3390/pharmaceutics13101612 The Indian Express (2021). Cuba says second COVID-19 vaccine Soberana 2 boasts
PMLiVE (2021). GSK/SK Bioscience COVID-19 Vaccine to Enter Phase 3 Trial. 91.2% efficacy. The Indian Express. Available online at: https://indianexpress.
Available online at: https://www.pmlive.com/pharma_news/gsksk_bioscience_ com/article/world/cuba-covid-19-vaccine-soberana-2-91-2-efficacy-7396330/
covid-19_vaccine_to_enter_phase_3_trial_1376104?SQ_ACTION=clear_ (accessed on Jun 11, 2022).
design_name&full=true (accessed on Sep 1, 2021). The Straits Times (2021). New Chinese vaccine shows 82% effectiveness against
Pourmalek, F., Rezaei Hemami, M., Janani, L., and Moradi-Lakeh, M. (2021). Rapid serious Covid-19 The Straits Times. Singapore: Singapore Press Holdings Ltd.
review of COVID-19 epidemic estimation studies for Iran. BMC Public Health Tian, J. H., Patel, N., Haupt, R., Zhou, H., Weston, S., Hammond, H., et al.
21:257. doi: 10.1186/s12889-021-10183-3 (2021). SARS-CoV-2 spike glycoprotein vaccine candidate NVX-CoV2373
Premkumar, L., Segovia-Chumbez, B., Jadi, R., Martinez, D. R., Raut, R., immunogenicity in baboons and protection in mice. Nat. Commun. 12:372.
Markmann, A. J., et al. (2020). The receptor-binding domain of the viral doi: 10.1038/s41467-020-20653-8
spike protein is an immunodominant and highly specific target of antibodies Toledo-Romani, M. E., Garcia-Carmenate, M., Silva, C. V., Baldoquin-Rodriguez,
in SARS-CoV-2 patients. Sci. Immunol. 5:eabc8413. doi: 10.1126/sciimmunol. W., Perez, M. M., Gonzalez, M. C. R., et al. (2021). Efficacy and Safety of
abc8413 SOBERANA 02, a COVID-19 conjugate vaccine in heterologous three doses
Reardon, S. (2021). Cuba’s bet on home-grown COVID vaccines is paying off. combination. medRxiv [Preprint]. doi: 10.1101/2021.10.31.21265703
Nature 600, 15–16. doi: 10.1038/d41586-021-03470-x Universo Online (2021). Cuba Inicia Nova Fase De Testes Com Vacina Que
Richmond, P. C., Hatchuel, L., Pacciarini, F., Hu, B., Smolenov, I., Li, P., et al. Desenvolve Contra Covid-19 (in Portuguese). São Paulo: Universo Online.
(2021). Persistence of the immune responses and cross-neutralizing activity Valdes-Balbin, Y., Santana-Mederos, D., Quintero, L., Fernandez, S., Rodriguez, L.,
with Variants of Concern following two doses of adjuvanted SCB-2019 COVID- Sanchez-Ramirez, B., et al. (2021). SARS-CoV-2 RBD-Tetanus toxoid conjugate
19 vaccine. J. Infect. Dis. 4:jiab447. vaccine induces a strong neutralizing immunity in preclinical studies. bioRxiv
Richmond, P., Hatchuel, L., Dong, M., Ma, B., Hu, B., Smolenov, I., et al. [Preprint]. doi: 10.1101/2021.02.08.430146
(2021). Safety and immunogenicity of S-Trimer (SCB-2019), a protein subunit Verma, N. (2021). Reassess, Reorganize & Realign Vision For Effective Inoculation:
vaccine candidate for COVID-19 in healthy adults: a phase 1, randomised, A Holistic Review Of Sars Covid-19 Vaccination Drive In India. Pandedih:
double-blind, placebo-controlled trial. Lancet 397, 682–694. doi: 10.1016/ Supremo Amicus.
S0140-6736(21)00241-5 Wang, N., Shang, J., Jiang, S., and Du, L. (2020). Subunit vaccines against emerging
Ryzhikov, A. B., Ryzhikov, E. À, Bogryantseva, M. P., Danilenko, E. D., Imatdinov, pathogenic human coronaviruses. Front. Microbiol. 11:298. doi: 10.3389/fmicb.
I. R., Nechaeva, E. A., et al. (2021). Immunogenicity and protectivity of the 2020.00298
peptide vaccine against SARS-CoV-2. Ann. Russ. Acad. Med. Sci. 76, 5–19. Wang, Q., Zhang, Y., Wu, L., Niu, S., Song, C., Zhang, Z., et al. (2020). Structural
doi: 10.15690/vramn1528 and functional basis of SARS-CoV-2 entry by using human ACE2. Cell 181,
Ryzhikov, A., Ryzhikov, E., Bogryantseva, M., Usova, S., Danilenko, E., Nechaeva, 894–904.e9. doi: 10.1016/j.cell.2020.03.045
E., et al. (2021). A single blind, placebo-controlled randomized study of the Yang, S., Li, Y., Dai, L., Wang, J., He, P., Li, C., et al. (2021). Safety and
safety, reactogenicity and immunogenicity of the “Epivaccorona” vaccine for immunogenicity of a recombinant tandem-repeat dimeric RBD-based protein

Frontiers in Microbiology | www.frontiersin.org 13 July 2022 | Volume 13 | Article 927306

Heidary et al. Protein Subunit Vaccines Against COVID-19

subunit vaccine (ZF2001) against COVID-19 in adults: two randomised, Publisher’s Note: All claims expressed in this article are solely those of the authors
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