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Volume 39 Issue 3
Mar.  2023
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WU Fang-hui, YIN Yi-feng, LIU Yan-li, . Preparation of nanobody against N protein of SARS-CoV-2 and assembly of colloidal gold immunochromatography card[J]. Chinese Journal of Public Health, 2023, 39(3): 389-393. doi: 10.11847/zgggws1140589
Citation: WU Fang-hui, YIN Yi-feng, LIU Yan-li, . Preparation of nanobody against N protein of SARS-CoV-2 and assembly of colloidal gold immunochromatography card[J]. Chinese Journal of Public Health, 2023, 39(3): 389-393. doi: 10.11847/zgggws1140589

Preparation of nanobody against N protein of SARS-CoV-2 and assembly of colloidal gold immunochromatography card

doi: 10.11847/zgggws1140589
  • Received Date: 2022-10-13
    Available Online: 2022-12-02
  • Publish Date: 2023-03-10
  •   Objective  To prepare nanobodies against N protein of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) and apply them to the detection of N protein antigen of SARS-COV-2.   Methods  The alpaca was immunized with N protein to construct a gene library of nanobody to N protein. The nanobody with affinity to N protein was screened with phage display technology; nanobody genes were cloned to yeast plasmids; positive clone strains were picked to induce expression; the secreted proteins were purified with affinity chromatography, desalted with dialysis and concentrated, then screened with enzyme-linked immunosorbent assay (ELISA) for the collection of high affinity nanobodies to N protein. A pair of nanobodies was screened with cross pairing experiment for the preparation of colloidal gold immunochromatography detection cards, and N-protein solution was dropwise added to the sample wells to observe the detection line and quality control line.   Results  Totally 46 nanobodies with different sequences were screened out and 6 of them were selected to express nanobodies with the concentration > 1.0 mg/ml and high affinity to N protein after purification, desalting and enrichment. Finally, the selected pair of antibodies was assembled into colloidal gold N protein immunodetection card to detect N protein with sandwich method. Obvious detection line and quality control line were observed in the test of SARS-CoV-2 N protein antigen using the detection card and the detection limit was ≥ 10 μg/mL for the test.   Conclusion  The prepared nanobody can be used in the colloidal gold nanobody immunochromatography detection card to detect SARS-CoV-2 N protein.
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    • Receive:  2022-10-13
    • Online:  2022-12-02
    • Published:  2023-03-10

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