Possible Cross-Reactivity of Feline and White-Tailed Deer Antibodies against the SARS-CoV-2 Receptor Binding Domain
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Abstract
We report cross-reactive antibodies from prepandemic cats and postpandemic South Carolina white-tailed deer that are specific for that SARS-CoV RBD. There are several potential explanations for this cross-reactivity, each with important implications to coronavirus disease surveillance.
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SciScore for 10.1101/2021.12.17.473265: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics not detected. Sex as a biological variable not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources After 1 hour, wells were washed 3x with PBS-T and a secondary antibody for that species was added (i.e., HRP goat-anti-human IgG, Rockland Immunochemicals, Pottstown, PA, USA; HRP goat-anti-dog IgG, Bethyl Laboratories, Montgomery, TX, USA; HRP goat-anti-cat IgG, Invitrogen, Waltham, MA, USA; HRP goat-anti-guinea pig IgG, Life Technologies Corp, Carlsbad, CA, USA; HRP rabbit-anti-deer IgG, KPL, Gaithersburg, MD, USA; HRP sheep-anti-cow IgG, Bethyl Laboratories, Montgomery, TX, USA) at … SciScore for 10.1101/2021.12.17.473265: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics not detected. Sex as a biological variable not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources After 1 hour, wells were washed 3x with PBS-T and a secondary antibody for that species was added (i.e., HRP goat-anti-human IgG, Rockland Immunochemicals, Pottstown, PA, USA; HRP goat-anti-dog IgG, Bethyl Laboratories, Montgomery, TX, USA; HRP goat-anti-cat IgG, Invitrogen, Waltham, MA, USA; HRP goat-anti-guinea pig IgG, Life Technologies Corp, Carlsbad, CA, USA; HRP rabbit-anti-deer IgG, KPL, Gaithersburg, MD, USA; HRP sheep-anti-cow IgG, Bethyl Laboratories, Montgomery, TX, USA) at dilutions of 1:10,000 (anti-human, cat, dog, tiger) or 1:250 ( IgGsuggested: Nonepig IgGsuggested: Noneanti-human , cat , dog , tigersuggested: NonePrimary antibodies of HRP conjugated mouse anti-6His (Proteintech, Rosemont, IL, USA) at 1:5,000 or polyclonal serum samples at 1:20 were incubated on blots at room temperature for 2 hours and subsequently washed 2x with TBS-T (Tris Buffered Saline with 0.1% Tween-20). anti-6Hissuggested: NoneExperimental Models: Cell Lines Sentences Resources Recombinant RBD purification: Supernatants from transfected HEK-293 T17 cells were collected into 50mL conical tubes and frozen at days 3 and 6 post-transfection. HEK-293 T17suggested: NoneRecombinant DNA Sentences Resources Vector pCAGGS contains the SARS-related coronavirus 2, Wuhan-Hu-1 pCAGGSsuggested: RRID:Addgene_127347)To produce recombinant RBD, the pCAGGS-RBD plasmid was transfected into ∼5×107 adherent HEK-293/T17 cells (ATCC CRL-11268) in a T-175 using PEI ( pCAGGS-RBDsuggested: NoneSoftware and Algorithms Sentences Resources OD450 values for each species and titration were graphed in GraphPad Prism ver. 8 GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)Sequences were viewed using 4Peaks software (Nucleobytes, Amsterdam, Netherlands) 4Peakssuggested: (4Peaks, RRID:SCR_000015)Sequences from fecal samples were Clustal W aligned to common coronaviruses, trimmed and a phylogenetic tree generated using Maximum-Likelihood method for each positive loci using MEGA X [58, 59]. MEGAsuggested: (Mega BLAST, RRID:SCR_011920)(GraphPad Software, San Diego, CA, USA). GraphPadsuggested: (GraphPad Prism, RRID:SCR_002798)Results from OddPub: Thank you for sharing your data.
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:- No conflict of interest statement was detected. If there are no conflicts, we encourage authors to explicit state so.
- No funding statement was detected.
- No protocol registration statement was detected.
Results from scite Reference Check: We found no unreliable references.
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