anti sars cov 2 spike protein test results interpretation

wxWd~{Trru%m#97Z=}<8boK.3E@KT>1oW#!7q%7uJ?IC5 .iM!. PubMed CAS Lancet Infect Dis 21, 13521354 (2021). x2- p` ` \0`0e`X{StAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA^@%&D7(mylkv You should not interpret the results of this test as an indication or degree of immunity or The titers were determined in duplicate assays from 5 mice in each group. UW MedicineDepartment of Laboratory MedicineVirology- Covid Testing Lab1601 Lind Ave SWRenton, WA 980573356Tel: (206) 685-6656 opt 4. DW, and MGA are named on patents that describe lipid nanoparticles for delivery of nucleic acid therapeutics, including mRNA and the use of modified mRNA in lipid nanoparticles as a vaccine platform. Centrifuge GOLD SST tube and route to Eastlake Virology (EVIR rack 81). Samples from 69 patients were included in this study. The results demonstrated that IgG2a/IgG1 (or Th1/Th2) ratios were greater than 1 in all vaccinated mice (Fig. The use of a specific threshold for decision-making regarding PreP in immunocompromised patients must be taken with cautions due to limitations, mainly based on the nature and type of assay used to measure the antibody. Characteristics like the number of bursts per electrode, their duration, frequency, and the number of spikes per burst according to the treatment condition were also quantified. The SARS-CoV-2 Spike IgG test shows the level of COVID-19 antibodies you had in your blood when you gave the blood sample. The 5-fold serially diluted mice sera were added in duplicate. Nanomaterial Delivery Systems for mRNA Vaccines. b hACE-2 binding assay (merged): culture supernatant collected from ChulaCov19 transfected cells incubated with HEK293T- hACE-2 cells. Hirabara, S. M. et al. Results are reported as AU/mL. The study also noted that the RBD may be accountable for the suppression of neuronal signals. Laboratoire AlphabioBiogroup, Marseille, France, Affiliation: Vaccines (Basel) 9, 850 (2021). The ChAdOx1 vectored vaccine, AZD2816, induces strong immunogenicity against SARS-CoV-2 beta (B.1.351) and other variants of concern in preclinical studies. The vaccine was measured for its immunogenicity in BALB/c mice both using ChulaCov19 alone or as heterologous prime/boost regimens alongside the approved vaccines (Fig. SARS-CoV-2 is an enveloped positive-sense single-stranded RNA beta coronavirus with a 30 kb polycistronic genome that encodes non-structural proteins (ORF1a and ORF1b, that are processed into Nsp1-16) at the 5-end, and structural proteins (S, E, M and N), and several other accessory factors (ORF3a . Heat-inactivated mice sera that were diluted 1:100 was added in duplicates into RBD-coated wells and incubated at RT for 2h. Then, either goat-antimouse IgG-HRP (1:40,000 dilution, KPL, USA) or goat-anti-mouse IgA-HRP (1:10,000 dilution, KPL, USA) was added to each well (100l/well) and incubated at RT for 1h. The peroxidase reaction was visualized by adding Sureblue TMB solution (KPL, USA) and incubating in the dark at RT for 15 and 20min for IgG and IgA ELISAs, respectively. 4e). Each dot represents an individual animal. Antibody escape of SARS-CoV-2 Omicron BA.4 and BA.5 from vaccine and BA.1 serum. Using a serologic test in combination with a NAAT to detect IgG or total antibodies 3 to 4 weeks after symptom onset maximizes the sensitivity and specificity to detect past SARS-CoV-2 infection. Adv. Since the outbreak of COVID-19, the world has raced to understand and accurately diagnose infection caused by SARS-CoV-2. In conclusion, whether it is generally believed that a certain level of antibodies is necessary to confer protection against the virus, but the exact level required is not yet clear. 6b). Recombinant S protein with S1/S2 cleavage site abolished (ACROBioSystems, China) was used as positive control both in HEK293T-hACE-2 binding assay and western blot. If testing will be delayed more than 7 days store at -20C or colder. Voysey, M. et al. [ view less ], Affiliations: the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Mean spike-specific IFN- positive T cells for 0.2, 1, 10 and 30g were 166, 429, 1913, and 1378 SFC/106 splenocytes, respectively. So there is not enough data available to comment on the uptake of this therapy yet and raises the question in cases of previous infection or vaccination, the need to assess the SARS-CoV-2 antibody level for therapy decision making [1820]. Using the Youden index maximization approach, optimal cutoffs were consistently lower than the reference cutoff of 264 BAU/ml for the Abbott and Beckman assays (148 and 48 BAU/ml, respectively). 2023. The S-specific total IgG after 1 or 2 doses of ChulaCov19 was analyzed in mice sera from experiment 1. INSTRUCTIONS FOR USE CoV2T Image Credit: whitehoune/Shutterstock.com. Baiersdorfer, M. et al. Immunogenicity and protective efficacy of SARS-CoV-2 mRNA vaccine Kim, H. W. et al. All assays showed a high AUC for prediction of positive and negative results of Genscript sVNT (AUC > 0.90 for all) (Fig 2). CAS With such promising results from animal studies, the same formulation of ChulaCov19 vaccine that had been tested in animals is currently in phase 1-2 of clinical trials and can be manufactured locally for later clinical development. Sci Transl Med 14, eabm3302 (2022). All p values <0.05 were defined as statistically significant. Neurological phenotypes induced by SARS-CoV-2 spike protein in neurons. Bars represent the GMTs and 95% CI for each group. As expected, Omicron subvariants, especially BA.4/5, showed the largest drop in micro-VNT50 titers (Fig. e0281257. Here we demonstrated that an LNP-encapsulated mRNA encoding a secreted form of prefusion nonstabilized ectodomain of SARS-CoV-2 spike protein ChulaCov19 was able to elicit robust, specific antibody and T-cell responses. On the basis of these data at present anti-SARS CoV-2 serological assays' results are not interchangeable, and, more importantly, individual immune monitoring should be performed with the same method. When compared with psVNT50 titers against BA.1, the GMT reduction against BA.4/5 in 10 and 30g dosed groups were 48 and 2.3 folds, respectively. Bloomberg. PubMed Evaluation of COVID-19 vaccine effectiveness in a changing landscape of COVID-19 epidemiology and vaccination: interim guidance, 1 October 2022: second addendum to Evaluation of COVID-19 vaccine effectiveness: interim guidance. Google Scholar. PubMed Central Molecular-based testing is used to diagnose COVID-19, and serologic testing of antibodies specific to SARS-CoV-2 is used to detect past infection. Watanabe, Y. et al. Biomedicines 10, 1464 (2022). 2a). The WHO International Standard for COVID-19 serological tests: towards In the immunogenicity dose-response and prime/boost studies (Experiment 1 and 2), NAb measurement was carried out as previously described56,68 based on live-virus micro-VNT50 against WT (Wuhan-Hu1), Alpha (B.1.1.7), Beta (B.1.351), Delta (B1.617.2) variants in VERO E6 cells with positive cut-off of 1:20. The encapsulated mRNA-LNP was characterized by various parameters including size, polydispersity (PDI) and mRNA encapsulation efficiency at 1, 6, and 12 months after manufacture. For example, for 10g dose group, the GMTs of psVNT50 for Delta (B.1.617.2) and Omicron (BA.1) variants decreased 5.9 and 14.3 folds when compared against WT (Wuhan-Hu1) strain, respectively (Fig. In all vaccinated groups, the number of spots that were detected after peptide pool #3-5 and pool #9 stimulation were 7484% and 810%, respectively (Fig. S0 was used to depict unprocessed S protein. Interim Guidelines for COVID-19 Antibody Testing | CDC In the present study, researchers quantified the neurological phenotypes induced in neurons by the SARS-CoV-2 S protein. SARS2Mutant: SARS-CoV-2 amino-acid mutation atlas database This test should not be used to diagnose or exclude acute SARS-CoV-2 infection. Labcorp test details for SARS-CoV-2 Semi-Quantitative Total Antibody, Spike . Cannabis users with a genetic predisposition to schizophrenia more likely to experience psychotic symptoms. Further information on research design is available in theNature Portfolio Reporting Summary linked to this article. Kunkalikar, Bhavana. Objectives: The aim was to determine the antibody response against SARS-CoV-2 spike protein and nucleoprotein using four automated immunoassays and three ELISAs for the detection of total Ig antibodies (Roche) or IgG (Abbott, Diasorin, Snibe, Euroimmun, Mikrogen) in COVID-19 patients. In brief, 100ng of recombinant S-trimer (ACROBioSystems, China) were coated to the 96-well plates. An mRNA Vaccine against SARS-CoV-2 - Preliminary Report. The team assessed the data using an algorithm devised in-house. Ying, B. et al. Moreover, anamnestic NAb response is undetectable in challenge animals. The National Library of Medicine is running a pilotto include preprints that result from research funded by NIH in PMC and PubMed. Nature 589, 603607 (2021). a Kinetics of total IgG at 2 weeks after receiving 1 or 2 doses of 0.2, 1, 10, and 30g of ChulaCov19. Follow-up testing with a molecular diagnostic should be considered to rule out infection in these individuals. Google Scholar. World Health Organization. World Health Organization (2022). However, this was still far lower than using homologous ChulaCov19 or AZD1222-prime/ChulaCov19-boost immunization regimens (Fig. Inclusion criteria were data from immunocompromised patients undergoing chemotherapy and/or biotherapy, aged over 18, who received at least one dose of the Pfizer/BioNTech BNT162b2 or Moderna COVID-19 vaccine (Spikevax) from three to six months before sampling collection. A. In a recent study posted to the bioRxiv* preprint server, researchers explored the association between severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein and burst activities in neurons. In addition, the pseudovirus neutralization test (psVNT50) against lentiviral pseudovirus bearing a codon-optimized spike gene, described previously69,70, was also used for determination of the neutralizing activity against WT, (Wuhan-Hu1), Alpha (B.1.1.7), Beta (B.1.351), Delta (B1.617.2), and Omicron (B1.1.529; BA.1 and BA.4/5 subvariants) variants. If material is not included in the articles Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. This contrasts with SARS CoV-1 where peak viral shedding occurs after patients were already quite ill5,6. Then, HRP-conjugated secondary antibodies, including rabbit anti-mouse IgG, dilution 1:10,000 (KPL, MD, USA), -IgG1 (dilution 1:5000), or -IgG2a dilution 1:5000 (both were from Southern Biotech, AL, USA) were added for an additional 1h. After washing, the signals were detected by adding tetramethylbenzidine (TMB) substrate (BioLegend, San Diego, CA, USA). Bleeding was performed at 2 weeks following each dose (and at week 18 for Experiment 3). This study aimed to describe serum-IgG responses to SARS-CoV-2 in a cohort of patients with both severe and mild COVID-19, including extended studies of patients who remained seronegative more than 90 . & Liu, J. Immunogenicity and safety of heterologous versus homologous prime-boost schedules with an adenoviral vectored and mRNA COVID-19 vaccine: a systematic review. All samples were collected at the Alphabio Laboratory in Marseille, France (European Hospital, AlphabioBiogroup). mRNA capping was performed by the trinucleotide cap1 analog, CleanCap (TriLink Biotechnologies, San Diego, CA, USA). Beckman assay showed lower values as compared to all other assays (P< 0.008 for all paired comparisons); and lower values was observed for Siemens assay compared with Roche assay (P = 0.0033). The procedure of mouse IFN- ELISPOT used in this study was described in our previous reports56,72. This research focuses on the impacts of the S protein. with these terms and conditions. PLOS ONE promises fair, rigorous peer review, However, there was no discernible difference in burst activity between S1-treated and the control wells. Comparative immunogenicity and reactogenicity of heterologous ChAdOx1-nCoV-19-priming and BNT162b2 or mRNA-1273-boosting with homologous COVID-19 vaccine regimens. 1a). If testing will be delayed more than 7 days store at -20C or colder. Signals of S protein stained by RBD-, S1-, S2-specific antibodies or PCS were detected on unpermeabilized HEK293T-hACE-2 cell after incubation with transfected supernatant. CK was also funded by emerging Infectious Diseases and Vaccines Cluster, Ratchadapisek Sompoch Endowment Fund (2021), Chulalongkorn University (764002-HE04), and the Second Century Fund (C2F), Chulalongkorn University and Ratchadapiseksompotch Fund. Lipid nanoparticles). Li, R. et al. This study was performed using sera collected between October 2021 and December 2021 from a real life cohort of 69 individuals attending internal medicine and infectious diseases department of the European Hospital (Marseille). Bars represent the meanSD of S-specific IFN- positive T cells after stimulated with overlapping peptide pools spanning the SARS-CoV-2 S1 (pooled #1-5) and S2 (pooled #6-10). It is notable that while all mice, except for one, dosed with 10-g and 1-g ChulaCov19 showed no detectable SARS-CoV-2 viral RNA in tested tissues. KR, DW, MGA, CK, EP, and SB are co-inventors of the submitted ChulaCov19 mRNA vaccines Patent. The bands corresponding to S1, S2 and intact S (S0) were detected. van Doremalen, N. et al. In contrast, CoronaVac immunization showed the lowest T cells responses (42 SFC/106 splenocytes). In this interview conducted at Pittcon 2023 in Philadelphia, Pennsylvania, we spoke to Dr. Chad Merkin, Director of the International Institute for Nanotechnology, about his work developing next-generation nanomaterials for medical applications. The geometric mean titers (GMTs) of NAb against wild-type (WT, Wuhan-Hu1) virus are 1,280, 11,762, 54,047, and 62,084, respectively. The comparable molecular weight of S0 expressed by ChulaCov19 was also observed when using commercial recombinant S with S1/S2 cleavage site abolished as control (Fig. SARS-CoV-2 Antibodies (NCVIGG, NCVIGQ)[NCVIGB], The qualitative detection of anti-Nucleocapsid IgG (NCVIGG) and the quantitative detection of anti-Spike IgG (NCVIGQ) antibodies. It is still being studied how does the immune system react in immunocompromised individuals, and how these observations translate into protection. These tests should not be used to diagnosis or exclude acute SARS-CoV-2 infection. Before administering S1 to neurons on day zero, a human monoclonal anti-S1 antibody was sampled and neutralized using the antibody. https://www.biorxiv.org/content/10.1101/2023.04.24.538161v1, Pregnant women show unique immune response to COVID-19, Study indicates that SARS-CoV-2 has evolved to gain increased replicative fitness and become well-adapted in epithelial cells of human airways, High-protein diet counters adaptive thermogenesis in prediabetic individuals. 5a). Competing interests: The authors have declared that no competing interests exist. Source data are provided as a source data file. Beyond the techniques used for the viral detection, the inverse correlation between vaccine dosage and tissue viremia might be the results of the quality of T cell response induced by the high vaccine dosage. PubMed All patients had received at least one dose of either Pfizer/BioNTech BNT162b2 or Moderna COVID-19 vaccine (Spikevax): 60 patients received Pfizer vaccine (87%) and 9 received Moderna vaccine (13%). Source data are provided with this paper. World Health Organization. Optimal cutoffs for distinguishing positivity were calculated using logistic regression on Genscript sVNT binary results (negative/positive), prior to the Youden index maximization approach on receiver operating characteristic curve results. Bar-On, L. et al. (2023) Anti-spike protein to determine SARS-CoV-2 antibody levels: Is there a specific threshold conferring protection in immunocompromised patients? Retrieved on May 01, 2023 from https://www.news-medical.net/news/20230427/Neurological-phenotypes-induced-by-SARS-CoV-2-spike-protein-in-neurons.aspx. In response to the COVID19 pandemic and in preparation for future pandemics, Thailand has funded this mRNA vaccine development program from preclinical to manufacturing and clinical development. Overall, the rescue experiment provided compelling evidence that S1 was able to suppress burst activities when exposed to cells early in their developmental course. sample is stable for 7 days at 2-8C once separated from a clot or red blood cells, or in a gel separator tube. By Day 4 after challenge, two mice in PBS-receiving group (control) began to show clinical signs of anorexia, lethargy, and rough hair coat. Antibody Testing After the COVID-19 Vaccine: What to - CreakyJoints Alene, M. et al. Liu, L. et al. Cellular and humoral responses after second and third SARS-CoV-2 Helmy, Y. Statistical significance was determined by two-sided MannWhitney test. Guilhem Cavaille, During the experiments, mice were maintained at 2022C and a relative humidity of 4510% on a 12h light/dark cycle. Previous study by Eichinger KM, et al. Lancet 396, 19791993 (2021). Comparing the clinical efficacy of COVID-19 vaccines: a systematic review and network meta-analysis. The OD450 of blanks were subtracted from OD450 of each sample before calculating antibody titer. Prompetchara, E. et al. Additional group (Experiment 3) immunized with 5g of ChulaCov19 was included for evaluation of NAb durability as measured at week 18 (15 weeks after received the 2nd dose) and the boosting effect of 3rd ChulaCov19 dose administered at week 20. JAMA Netw Open 4, e2137257 (2021). SARS-CoV-2 Testing | COVID-19 Treatment Guidelines The Abbott AdviseDx SARS-CoV-2 IgG II immunoassay detects antibodies to the viral spike protein (S). News-Medical, viewed 01 May 2023, https://www.news-medical.net/news/20230427/Neurological-phenotypes-induced-by-SARS-CoV-2-spike-protein-in-neurons.aspx. Five microliters of each RNA sample was used in quantitative RT-PCR that was performed using CDC procedure73 and AFRIMS SOPs in vitro SARS-CoV-2 RNA transcripts (IVTs). Your Spike Protein Antibody results will be reported as a reference range: >/= 0.80 U/mL: This is a positive result for anti-SARS CoV-2S. Chutitorn Ketloy. A subgroup analysis was performed according to vaccination status (complete or incomplete). This is consistent with a previous report46. Tian, J. H. et al. ];V^srE]DwCyTPre_fyG;Cb@*\d$ j '-|,!]jF/J9r\s%3! S-specific IgG measurement was performed employing indirect ELISA as described previously56,67. 2a). ChAdOx1 nCoV-19 (AZD1222) or nCoV-19-Beta (AZD2816) protect Syrian hamsters against Beta Delta and Omicron variants. This was followed by exposure to the same S1 concentration for seven consecutive days. The proprietary lipid and LNP composition are described in patent application WO2020097540A161,62. IgG2a and IgG1 subclasses were also assessed to determine Th1 and Th2 responses, respectively. In Experiment 3, the durability of NAb induced by ChulaCov19 was monitored until week 18 (15 weeks after the 2nd dose). ACS Cent Sci 7, 594602 (2021). 11 Antibody tests may help identify past SARS-CoV-2 infection if performed two to four. Sera were collected at weeks 0, 2, 3, 4+6 days, and 5+6 days for NAb measurements. Her college project work based on The manifestations and causes of sickle cell anemia formed the stepping stone to a life-long fascination with human pathophysiology. Available from: https://www.science.org/content/article/new-crop-covid-19-mrna-vaccines-could-be-easier-store-cheaper-use (2022). S protein on HEK293T-hACE-2 cell surface was stained with the same antibodies used in 2a. Note: tissues from 3/5 animals in control group were collected at day 5. Goat-anti-mouse IgG-FITC, donkey-anti-rabbit IgG-FITC (both were from BioLegend, CA, USA) or goat-anti-human AlexaFluor647 (Southern Biotech, AL, USA), at dilution of 1:5,000 was used as secondary antibodies following anti-RBD, -S1, -S2 or PCS staining. A. 1b. : draft manuscript preparation. SARS-COV-2 Variants: Differences and Potential of Immune Evasion. At week 18, the NAb against WT (Wuhan-Hu1) and Delta (B.1.617.2) decreased approximately 2-fold but not statistically significant when compare with week 5 titers. https://ClinicalTrials.gov/show/NCT05231369 (2022). S protein on HEK293T-hACE-2 was stained with anti-RBD, -S1, -S2 or PCS and detected using the same procedure described above. Protein OCLN found to play crucial role in SARS-CoV-2 cell-to-cell transmission, Study reveals survival time of SARS-CoV-2 in wastewater: Implications for public health, The BCG vaccine does not decrease the risk of COVID-19 in healthcare workers. In this study, the S1 and S2 subunits of the spike protein were evaluated separately to determine if they elicited any neurological phenotypes as estimated by the micro-electrode arrays (MEAs). Prefusion RSV F Immunization Elicits Th2-Mediated Lung Pathology in Mice When Formulated With a Th2 (but Not a Th1/Th2-Balanced) Adjuvant Despite Complete Viral Protection. S1 neutralized by antibodies did not result in a significant decrease in burst activity compared to the control, whereas the conventional S1 treatment on day zero did reduce burst activity. Klemis, V. et al. Stability: Sample stable off the clot, red blood cells, or separator gel for 7 days at 2-8C. Global Percentage of Asymptomatic SARS-CoV-2 Infections Among the Tested Population and Individuals With Confirmed COVID-19 Diagnosis: A Systematic Review and Meta-analysis. In mice, ChulaCov19 was highly immunogenic as a booster in settings primed with either inactivated or viral vector vaccine. For SARS-CoV-2, tests to neutralize live viruses are performed only in specialized laboratories and are not standardized, making it difficult to compare and justify the use of a well-characterized sVNT as a functional reference [24,25].Additionally, neutralizing antibodies were not investigated, which could have helped in determining whether the anti-RBD or the anti-spike assays had the strongest correlation with virus neutralization. Article Six-day post challenge, wk5+6 days, mice were sacrificed to determine virus titers in different tissues (nasal turbinate, brain, lung, and kidney) and for histopathology. At 2104 PFU of SARS-CoV-2 inoculum, PBS-vaccinated mice displayed clinical symptoms or weight loss within 1 day and all mice succumbed by day 6. The Abbott SARS-CoV-2 IgG immunoassay detects antibodies to the viral nucleocapsid protein (NP). The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of coronavirus disease 2019 (COVID-19). Available from: https://covid19.trackvaccines.org/agency/who (2022). They concluded that higher levels of all immune markers were correlated with a reduced risk of symptomatic infection. PLoS One 16, e0248007 (2021). At week 5 (2 weeks after the second dose), all mice in both vaccinated groups showed increased NAb levels. Correspondence to Sign up for the Nature Briefing newsletter what matters in science, free to your inbox daily. "Neurological phenotypes induced by SARS-CoV-2 spike protein in neurons". The differences in the commercial assays used in this study are related to the components of the tests (the spike antigen epitopes used, the different isolates of the SARS-CoV-2, and the quantification of either total antibodies or only IgG) [2123]. Experiment 1: (a) Live-virus microneutralization (micro-VNT50) titers against WT (Wuhan-Hu1) live-virus at two weeks after receiving each vaccine dose. Posted in: Medical Science News | Medical Research News | Disease/Infection News, Tags: ACE2, Angiotensin, Angiotensin-Converting Enzyme 2, Antibodies, Antibody, Blood, Blood Pressure, Brain, Cell, Coronavirus, Coronavirus Disease COVID-19, covid-19, Electrode, Enzyme, Frequency, Membrane, micro, Neurons, Newborn, Phenotype, Protein, Receptor, Research, Respiratory, SARS, SARS-CoV-2, Severe Acute Respiratory, Severe Acute Respiratory Syndrome, Spike Protein, Syndrome, Vaccine, Virus. Derby, M., Alexander-Miller, M., Tse, R. & Berzofsky, J. High-avidity CTL exploit two complementary mechanisms to provide better protection against viral infection than low-avidity CTL. Cell 182, 5058.e58 (2020). Slider with three articles shown per slide. All studies were conducted under protocols approved by the Committees on Care of Laboratory Animal Faculty of Medicine, Chulalongkorn University (IACUC approval no. Thus, most recently approved vaccines employ full-length S (with or without modification) or whole virus (inactivated) as a target antigen19. In the same study, two doses of AZD1222 could protect rhesus macaque form viral challenge. showed time-dependent changes in the comparability of different antibody tests with samples collected at different time points [26]. as a booster dose in mice that had been primed with CoronaVac or AZD1222 (Experiment 2). ChulaCov19 significantly enhanced the magnitude of both NAb and T cell responses compared to homologous 2-dose regimens of either CoronaVac or AZD1222. Substantial undocumented infection facilitates the rapid dissemination of novel coronavirus (SARS-CoV-2). 399, 924944 (2022). Science 368, 489493 (2020). Goat-anti-human IgG, goat-anti-mouse IgG, or goat-anti-rabbit IgG antibodies (all were diluted 1:10,000) conjugated with horseradish peroxidase (HRP) were used as secondary antibodies (all were from KPL, MD, USA) and detected by chemiluminescence substrate (Immobilon western, Millipore, CA, USA) then exposed to an X-ray film. S-specific total IgG analyzed at week 2 revealed that all ChulaCov19-immunized mice, either with 1 or 2 doses, elicited anti-S-specific IgG response from the lowest dose of 0.2g with a dose-dependent response pattern. Safety and Efficacy of the BNT162b2 mRNA Covid-19 Vaccine. The second dose of ChulaCov19 strongly augmented the IgG antibody levels with an increase of 10-19 folds, p<0.01 for all dose ranges (Fig. Two were quantitative: Abbott SARS-CoV-2 IgG II Quant-test (Abbott) (Abbott France, Rungis, France) with 50 arbitrary units (AU)/ml as a threshold for positivity, and Roche Elecsys anti-SARS-CoV-2 S (Roche Diagnostics France, Meylan, France) with 0.8 AU/ml used as a threshold for positivity. Note; 4 mice in 10g group were analyzed for psVNT50 against BA.4/5 due to the limited volume of serum samples. Vaccine inequity issue remains a major global challenge. On Day 5, significant weight reduction (p<0.05) was observed in control group when compared with the vaccinated groups. Pharmaceutics 14, 1427 (2022). SARS-CoV-2 neutralizing antibodies decline over one year and patients with severe COVID-19 pneumonia display a unique cytokine profile. Briefly, 100ng/well of RBD recombinant proteins (Abcam, UK) were coated overnight to the 96-well plates. (accessed May 01, 2023). Experiment 1: dose-response of homologous ChulaCov19 prime/boost study, mice were immunized twice intramuscularly at 3 weeks interval of ChulaCov19 with dosage ranging from 0.2, 1, 10, to 30g. Three out of five mice reached euthanasia criteria on Day 5, and symptoms progressed for the remaining two mice which met the criteria on Day 6. Discover a faster, simpler path to publishing in a high-quality journal. The authors would like to thanks Dr.Navapon Techakriengkrai, Faculty of Veterinary Science, Chulalongkorn University for providing HEK293T-hACE-2 cells.

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