Preparation and identification of a broad-spectrum antibody targeting hemagglutinin protein of influenza A virus
10.3760/cma.j.cn112309-20211119-00393
- VernacularTitle:靶向甲型流感病毒血凝素蛋白的广谱抗体制备及鉴定
- Author:
Nan CHEN
1
;
Lihui CHAI
;
Guojiang CHEN
;
Jing WANG
;
Chunxia QIAO
;
Xinying LI
;
Chenghua LIU
;
Beifen SHEN
;
Jiannan FENG
;
He XIAO
Author Information
1. 军事科学院军事医学研究院毒物药物研究所，北京 100850
- Keywords:
Influenza A virus;
Hemagglutinin;
Broad spectrum;
Neutralizing activity
- From:
Chinese Journal of Microbiology and Immunology
2022;42(8):636-641
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To prepare and identify a broad-spectrum antibody FHA3 targeting influenza A virus hemagglutinin (HA).Methods:According to the single-chain antibody fragment (scFv) sequence, the heavy chain (VH) and light chain (VL) variable regions of FHA3 were amplified by PCR and a recombinant plasmid pFRT-IgG1κ-FHA3 was constructed by linking the expression vector pFRT-IgG1κ. FHA3 was expressed in the ExpiCHO system and purified by affinity purification. The binding activity of FHA3 to influenza A virus HA was detected by ELISA. The neutralizing activity of FHA3 was detected in vitro by infecting host cells with pseudovirus. Results:SDS-PAGE showed that high-purity FHA3 was obtained. FHA3 could bind to H1N1 HA, H2N2 HA, H3N2 HA, H5N1 HA, H7N9 HA and H9N2 HA in a concentration-dependent manner. FHA3 had good neutralizing activity in vitro that was it could effectively block the invasion of H5N1 and H7N9 pseudoviruses into target cells at a low concentration of 5 μg/ml and H1N1 pseudovirus at 0.012 5 μg/ml. Conclusions:A broad spectrum antibody targeting HA protein of influenza A virus with neutralizing activity in vitro was obtained.
