Defining Factors that Influence vaccine-induced, cross-variant neutralizing antibodies for SARS-CoV-2 in Asians

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   ScreenIT

   Mar 11, 2022

   SciScore for 10.1101/2022.03.06.22271809: (What is this?)

   Please note, not all rigor criteria are appropriate for all manuscripts.

   Table 1: Rigor

   Ethics	Consent: All participants provided written informed consent in accordance with the Declaration of Helsinki for Human Research. IRB: Ethics committee of National Healthcare Group (NHG) Domain Specific Review Board (DSRB) Singapore gave ethical approval for this work.
   Sex as a biological variable	not detected.
   Randomization	All concentrations were tested three times at randomized orders.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Quantitative ELISA: Concentrations of anti-Spike, anti-RBD, and anti-Nucleocapsid IgG antibodies in plasma samples were estimated using quantitative ELISA39.	anti-Spike , anti-RBD , sugge…

   SciScore for 10.1101/2022.03.06.22271809: (What is this?)

   Please note, not all rigor criteria are appropriate for all manuscripts.

   Table 1: Rigor

   Ethics	Consent: All participants provided written informed consent in accordance with the Declaration of Helsinki for Human Research. IRB: Ethics committee of National Healthcare Group (NHG) Domain Specific Review Board (DSRB) Singapore gave ethical approval for this work.
   Sex as a biological variable	not detected.
   Randomization	All concentrations were tested three times at randomized orders.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Quantitative ELISA: Concentrations of anti-Spike, anti-RBD, and anti-Nucleocapsid IgG antibodies in plasma samples were estimated using quantitative ELISA39.	anti-Spike , anti-RBD , suggested: None anti-Nucleocapsid IgG suggested: (SICGEN Cat# AB0369, RRID:AB_2895400)
   To estimate the concentrations of anti-spike and anti-RBD antibodies, an RBD-specific human monoclonal IgG antibody named LSI-COVA-015 isolated from COVID-19 convalescent patient was diluted to a series of concentrations ranging from1 ng/mL to 1 µg/mL and added at 100 µL/well.	anti-RBD suggested: None
   Similarly, a nucleocapsid-specific monoclonal IgG antibody named LSI-COVANC-D generated from hybridoma cloning40,41 was added at the same concentrations to estimate the concentration of anti-nucleocapsid antibodies in the plasma samples.	anti-nucleocapsid suggested: None
   After 60-minutes incubation, plate was washed and incubated with goat anti-human IgG-HRP antibodies (Invitrogen #31413, diluted 10000-times in blocking buffer) at 100 µL/well for 50 minutes, protected from light.	anti-human IgG-HRP suggested: None
   Standard curves were constructed using the reference antibodies from 100 ng/mL to 1 ng/mL, and the concentration of antigen-specific IgG antibodies in plasma samples were calculated via interpolation.	antigen-specific IgG suggested: None
   Quantification of serological IgG antibodies against SARS-CoV-2 antigens by ELISA.	SARS-CoV-2 suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   A total of 36 ×106 HEK293T cells were transfected with 27 µg pMDLg/pRRE (Addgene, #12251), 13.5 µg pRSV-Rev (Addgene, #12253), 27 µg pTT5LnX-WHCoV-St19 (	HEK293T suggested: None
   Pseudovirus neutralization test (PVNT): ACE2 stably expressed CHO cells were seeded at a density of 5×104 cells in 100 µL of complete medium [DMEM/high glucose with sodium pyruvate (Gibco, #10569010), supplemented with 10% FBS (Hyclone, #SV301160.03),10	CHO suggested: None
   Recombinant DNA
   Sentences	Resources
   Gene encoding SARS-CoV-2 Nucleocapsid (Biobasic) was cloned into pNIC28 and expressed in BL21(	pNIC28 suggested: RRID:Addgene_125591)
   A total of 36 ×106 HEK293T cells were transfected with 27 µg pMDLg/pRRE (Addgene, #12251), 13.5 µg pRSV-Rev (Addgene, #12253), 27 µg pTT5LnX-WHCoV-St19 (	pMDLg/pRRE suggested: RRID:Addgene_12251) pRSV-Rev suggested: RRID:Addgene_12253) pTT5LnX-WHCoV-St19 suggested: None
   SARS-CoV-2 Spike) and 54 µg pHIV-Luc-ZsGreen (Addgene, #39196) using Lipofectamine 3000 transfection reagent (Invitrogen, #L3000-150) and cultured in a 37 °C, 5% CO2 incubator for three days.	pHIV-Luc-ZsGreen suggested: RRID:Addgene_39196)
   Software and Algorithms
   Sentences	Resources
   Ethics committee of National Healthcare Group (NHG) Domain Specific Review Board (DSRB) Singapore gave ethical approval for this work.	National Healthcare suggested: None
   All analyses were performed using Graphpad Prism 9.0 and can be found in Supplementary Data.	Graphpad Prism suggested: (GraphPad Prism, RRID:SCR_002798)

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   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).

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   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.

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   Results from TrialIdentifier: No clinical trial numbers were referenced.

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   Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

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   Results from JetFighter: We did not find any issues relating to colormaps.

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   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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   Results from scite Reference Check: We found no unreliable references.

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   Version published to 10.1101/2022.03.06.22271809v1 on medRxiv

   Mar 8, 2022