Generation of antibodies against COVID-19 virus for development of diagnostic tools

  1. Maohua Li
  2. Ronghua Jin
  3. Ya Peng
  4. Cuiyan Wang
  5. Wenlin Ren
  6. Fudong Lv
  7. Sitao Gong
  8. Feng Fang
  9. Qianyun Wang
  10. Jianli Li
  11. Tong Shen
  12. Hunter Sun
  13. Lei Zhou
  14. Yali Cui
  15. Hao Song
  16. Le Sun

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Abstract

The COVID-19 China coronavirus started in Dec 2019 was challenged by the lack of accurate serological diagnostic tool for this deadly disease to quickly identify and isolate the infected patients. The generation of COVID-19-specific antibodies is essential for such tasks. Here we report that polyclonal and monoclonal antibodies were generated by immunizing animals with synthetic peptides corresponding to different areas of Nucleoprotein (N) of COVID-19. The specificities of the COVID-19 antibodies were assessed by Western Blot analysis against NPs from COVID-19, MERS and SARS. Antibodies were used for immunohistochemistry staining of the tissue sections from COVID-19 infected patient, as a potential diagnostic tool. A Sandwich ELISA kit was quickly assembled for quantitation of the virus/NP of COVID-19 concentrations in the vaccine preparations. Development of POCT is also aggressively undergoing.

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  1. ScreenIT

    SciScore for 10.1101/2020.02.20.20025999: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board Statementnot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Experimental Models: Cell Lines
    SentencesResources
    Splenocytes from the immunized mice were fused with the mouse myeloma cell line SP2/0 and cultural supernatant from individual hybridoma clones were screened against NP by ELISA.
    SP2/0
    suggested: CLS Cat# 400481/p8829_Sp20-Ag14, RRID:CVCL_2199)
    Experimental Models: Organisms/Strains
    SentencesResources
    Balb/c mice and New Zealand White rabbits were immunized with KLH-conjugated synthetic peptides.
    Balb/c
    suggested: RRID:IMSR_ORNL:BALB/cRl)
    Software and Algorithms
    SentencesResources
    Peptides synthesis and antibody generation: Peptides were synthesized at DentriPro Bioscientific Ltd.
    DentriPro Bioscientific
    suggested: None

    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.

    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.

  2. Version published to 10.1101/2020.02.20.20025999 on medRxiv