Antibodies to detect novel coronavirus
A coronavirus and antibody technology, applied in the fields of molecular biology and cellular immunology, can solve the problems of missed diagnosis, false positive and false negative, and large impact
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Embodiment 1
[0086] Example 1 Antibody Screening
[0087] 1. Expression of recombinant SARS-CoV2 nucleoprotein (NP)
[0088] 1.1 Main reagents
[0089] The SARS-CoV2 NP gene sequence (GenBank serial number: MT066176.1) and related primers were synthesized and sequenced by General Biosystems (Anhui) Co., Ltd.; E.coli DH5α, BL21(DE3) competent cells were purchased from General Biosystems (Anhui) Co., Ltd.; BamHI and NotI endonucleases were purchased from New England Biolabs (NEB); EXTaq enzyme was purchased from TaKaRa; HRP-labeled anti-human Fc antibody was purchased from Sigma; other chemical reagents were domestic analytical reagents ;The serum of patients infected with 2019-nCoV is collected and preserved by our center.
[0090] 1.2 Prokaryotic expression plasmid construction
[0091] The primers for prokaryotic expression of NP gene were designed, the upstream primer had a BamHI restriction site, and the downstream primer had a NotⅠ restriction site. The primer sequences are: Cov2-N...
Embodiment 2
[0142] Example 2 Expression of whole antibody and related functional verification
[0143] From the obtained 159 antibodies, 16 antibodies were finally selected for the expression of the whole antibody and related functional verification, and these 16 antibodies were named JS01-JS16.
[0144] Among them, the JS05 antibody sequence is as follows:
[0145] The amino acid sequence of the heavy chain variable region CDR1 is shown in SEQ ID NO.1;
[0146] The amino acid sequence of the heavy chain variable region CDR2 is shown in SEQ ID NO.2;
[0147] The amino acid sequence of the heavy chain variable region CDR3 is shown in SEQ ID NO.3;
[0148] The amino acid sequence of the light chain variable region CDR1 is shown in SEQ ID NO.5;
[0149] The amino acid sequence of the light chain variable region CDR2 is shown in SEQ ID NO.6;
[0150] The amino acid sequence of the light chain variable region CDR3 is shown in SEQ ID NO.7;
[0151] The amino acid sequence of the heavy chai...
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