Identification of IgG antibody response to SARS-CoV-2 spike protein and its receptor binding domain does not predict rapid recovery from COVID-19
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Abstract
Diagnostic testing and evaluation of patient immunity against the novel severe acute respiratory syndrome (SARS) corona virus that emerged last year (SARS-CoV-2) are essential for health and economic crisis recovery of the world. It is suggested that potential acquired immunity against SARS-CoV-2 from prior exposure may be determined by detecting the presence of circulating IgG antibodies against viral antigens, such as the spike glycoprotein and its receptor binding domain (RBD). Testing our asymptomatic population for evidence of COVID-19 immunity would also offer valuable epidemiologic data to aid health care policies and health care management. Currently, there are over 100 antibody tests that are being used around the world without approval from the FDA or similar regulatory bodies, and they are mostly for rapid and qualitative assessment, with different degrees of error rates. ELISA-based testing for sensitive and rigorous quantitative assessment of SARS-CoV-2 antibodies can potentially offer mechanistic insights into the COVID-19 disease and aid communities uniquely challenged by limited financial resources and access to commercial testing products. Employing recombinant SARS-CoV-2 RBD and spike protein generated in the laboratory, we devised a quantitative ELISA for the detection of circulating serum antibodies. Serum from twenty SARS-CoV-2 RT-PCR confirmed COVID-19 hospitalized patients were used to detect circulating IgG titers against SARS-CoV-2 spike protein and RBD. Quantitative detection of IgG antibodies to the spike glycoprotein or the RBD in patient samples was not always associated with faster recovery, compared to patients with borderline antibody response to the RBD. One patient who did not develop antibodies to the RBD completely recovered from COVID-19. In surveying 99 healthy donor samples (procured between 2017-February 2020), we detected RBD antibodies in one donor from February 2020 collection with three others exhibiting antibodies to the spike protein but not the RBD. Collectively, our study suggests that more rigorous and quantitative analysis, employing large scale samples sets, is required to determine whether antibodies to SARS-CoV-2 spike protein or RBD is associated with protection from COVID-19 disease. It is also conceivable that humoral response to SARS-CoV-2 spike protein or RBD works in association with adaptive T cell response to determine clinical sequela and severity of COVID-19 disease.
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SciScore for 10.1101/2020.05.01.20087684: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IRB: Serum from SARS-CoV-2 patients were obtained from the Memorial Hermann Hospital-Texas Medical Center (n=20) under the approval of Institutional Review board at the University of Texas McGovern Medical School at Houston. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Secondary antibodies used were goat anti-mouse HRP (W4028, Promega, 1:2000) and donkey anti-rabbit HRP (Ab16284, Abcam, 1:2000) in 2% BSA TBS/T, respectively. anti-mouse HRP ( W4028suggested: Noneanti-rabbit HRPsuggested: (Abcam Cat# ab16284, …SciScore for 10.1101/2020.05.01.20087684: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IRB: Serum from SARS-CoV-2 patients were obtained from the Memorial Hermann Hospital-Texas Medical Center (n=20) under the approval of Institutional Review board at the University of Texas McGovern Medical School at Houston. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Secondary antibodies used were goat anti-mouse HRP (W4028, Promega, 1:2000) and donkey anti-rabbit HRP (Ab16284, Abcam, 1:2000) in 2% BSA TBS/T, respectively. anti-mouse HRP ( W4028suggested: Noneanti-rabbit HRPsuggested: (Abcam Cat# ab16284, RRID:AB_955387)Ab16284suggested: (Abcam Cat# ab16284, RRID:AB_955387)For the performance assays shown in Figure 3c-d, 50 microliters of anti-StrepTagll antibody (Table S1, 1:500) in 2% BSA was added to the wells. anti-StrepTagllsuggested: NoneExperimental Models: Cell Lines Sentences Resources Cell culture and transfection: 293T/17 (HEK 293T/17) cells from ATCC (CRL-11268) were cultured in DMEM (Corning) with 10% fetal bovine serum (FBS, Gemini). HEK 293T/17suggested: ATCC Cat# CRL-11268, RRID:CVCL_1926)Software and Algorithms Sentences Resources Data presentation and statistical analyses: Statistical analysis and generation of the graphs were carried out using GraphPad Prism. GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).
Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.Results from TrialIdentifier: No clinical trial numbers were referenced.
Results from Barzooka: We did not find any issues relating to the usage of bar graphs.
Results from JetFighter: We did not find any issues relating to colormaps.
Results from rtransparent:- Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
- Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
- No protocol registration statement was detected.
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