Preparation and identification of antibodies targeting influenza A virus N1 subtype neuraminidase
10.3760/cma.j.cn112309-20220127-00033
- VernacularTitle:靶向甲型流感病毒N1亚型神经氨酸酶抗体的制备及鉴定
- 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;
Neuraminidase;
N1 subtype;
Pseudovirus release
- From:
Chinese Journal of Microbiology and Immunology
2022;42(11):851-856
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To prepare and identify a functional antibody FNA1 targeting the neuraminidase (NA) of influenza A virus N1 subtype.Methods:According to single-chain antibody fragment (scFv) sequence, the heavy chain and light chain variable region sequences of FNA1 were synthesized, and the recombinant expression plasmid pFRT-IgG1κ-FNA1 was constructed by linking the expression vector pFRT-IgG1κ. The FNA1 antibody was expressed in ExpiCHO cells and purified using affinity purification technique. The binding ability of FNA1 to the target proteins, influenza A virus N1 subtype NA antigens, was detected by ELISA. Flow cytometry was performed to analyze the binding ability of FNA1 to the NA antigens expressed on the surface of cell membrane. The in vitro activity of FNA1 against NA was evaluated by infecting 293T cells with pseudovirus. Results:Protein electrophoresis showed that FNA1 with high purity was obtained. FNA1 specifically recognized and bound to N1 subtype NA antigens in a concentration-dependent manner. FNA1 could effectively block NA activity by binding to N1 subtype NA protein expressed on the surface of cell membrane, thus inhibiting the release of packaged pseudovirus from cell surface and further inhibiting target cell infection.Conclusions:An antibody FNA1 targeting influenza A virus N1 subtype NA with in vitro functional activity was obtained.
