Monoclonal antibodies for the S2 subunit of spike of SARS-CoV-1 cross-react with the newly-emerged SARS-CoV-2
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Abstract
A novel coronavirus, SARS-CoV-2, which emerged at the end of 2019 and causes COVID-19, has resulted in worldwide human infections. While genetically distinct, SARS-CoV-1, the aetiological agent responsible for an outbreak of severe acute respiratory syndrome (SARS) in 2002–2003, utilises the same host cell receptor as SARS-CoV-2 for entry: angiotensin-converting enzyme 2 (ACE2). Parts of the SARS-CoV-1 spike glycoprotein (S protein), which interacts with ACE2, appear conserved in SARS-CoV-2.
Aim
The cross-reactivity with SARS-CoV-2 of monoclonal antibodies (mAbs) previously generated against the S protein of SARS-CoV-1 was assessed.
Methods
The SARS-CoV-2 S protein sequence was aligned to those of SARS-CoV-1, Middle East respiratory syndrome (MERS) and common-cold coronaviruses. Abilities of mAbs generated against SARS-CoV-1 S protein to bind SARS-CoV-2 or its S protein were tested with SARS-CoV-2 infected cells as well as cells expressing either the full length protein or a fragment of its S2 subunit. Quantitative ELISA was also performed to compare binding of mAbs to recombinant S protein.
Results
An immunogenic domain in the S2 subunit of SARS-CoV-1 S protein is highly conserved in SARS-CoV-2 but not in MERS and human common-cold coronaviruses. Four murine mAbs raised against this immunogenic fragment could recognise SARS-CoV-2 S protein expressed in mammalian cell lines. In particular, mAb 1A9 was demonstrated to detect S protein in SARS-CoV-2-infected cells and is suitable for use in a sandwich ELISA format.
Conclusion
The cross-reactive mAbs may serve as useful tools for SARS-CoV-2 research and for the development of diagnostic assays for COVID-19.
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SciScore for 10.1101/2020.03.06.980037: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. 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 Cells were immunolabelled for 1 hour at RT with the indicated murine mAb and 45 min with Alexa Fluor 488-conjugated goat anti-mouse IgG antibody (Life Technologies). anti-mouse IgGsuggested: NoneThey were then washed 3 times with PBS and incubated with goat anti-mouse conjugated to Alexa Fluor 488-conjugated goat anti-mouse IgG antibody (Thermo Fisher) containing DAPI in immunofluorescence buffer for an additional incubation of 30 minutes. anti-mousesuggeste…SciScore for 10.1101/2020.03.06.980037: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. 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 Cells were immunolabelled for 1 hour at RT with the indicated murine mAb and 45 min with Alexa Fluor 488-conjugated goat anti-mouse IgG antibody (Life Technologies). anti-mouse IgGsuggested: NoneThey were then washed 3 times with PBS and incubated with goat anti-mouse conjugated to Alexa Fluor 488-conjugated goat anti-mouse IgG antibody (Thermo Fisher) containing DAPI in immunofluorescence buffer for an additional incubation of 30 minutes. anti-mousesuggested: NoneExperimental Models: Cell Lines Sentences Resources Cells: Vero E6 and COS-7 cells were purchased from American Type Culture Collection (Manassas, VA, USA) and cultured in Dulbecco’s Modified Eagle’s Medium (DMEM; Thermo Scientific) supplemented with 10% foetal bovine serum (FBS; HyClone) and penicillin-streptomycin solution (Thermo Fisher Scientific). Vero E6suggested: NoneCOS-7suggested: NoneVero-E6 cells were infected with SARS-CoV-2 (SARS-CoV-2-Iso/01/human/2020/SWE, GenBank accession no. MT093571) at a multiplicity of infection (MOI) of 1 in DMEM 2% FBS (Thermo Fisher). Vero-E6suggested: NoneSoftware and Algorithms Sentences Resources A multiple sequence alignment was created with MAFFT using the slow but accurate L-INS-I parameter settings [21] and the alignment curated, cut to the target region 1029-1192 (SARS-CoV numbering) and visualized with Jalview [22]. MAFFTsuggested: (MAFFT, RRID:SCR_011811)Jalviewsuggested: (Jalview, RRID:SCR_006459)The nucleotide sequences were searched with BLASTX against the reference spike glycoprotein. BLASTXsuggested: (BLASTX, RRID:SCR_001653)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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