Basic characteristics of a human monoclonal antibody against rabies virus
10.3760/cma.j.cn112866-20200326-00092
- VernacularTitle:1株人源抗狂犬病病毒单克隆抗体的基本特征
- Author:
Jijun CHEN
1
;
Jianjun NAN
;
Xiaorui ZHAO
;
Ying XIONG
;
Chen AN
;
Xiaoyan MAO
Author Information
1. 兰州生物制品研究所有限责任公司第四研究室 730046
- Keywords:
Monoclonal;
Lyssavirus;
Epitope;
Neutralizing index;
Escape mutant virus;
Thermal stability
- From:
Chinese Journal of Experimental and Clinical Virology
2020;34(5):500-505
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To evaluate properties of monoclonal anti-rabies virus antibodies obtained from phage library.Methods:A positive clone was obtained from the phage library of anti-rabies virus, and the variable region of light and heavy chain of the antibody was amplified by PCR. The transient expression plasmid of the fully human antibody was constructed and then transiently expressed in HEK293 EBNA1 cells. The purified antibody was analyzed by in vitro anti-rabies virus neutralization activity, neutralization index, escape virus glycoprotein sequencing and thermal stability by differential scanning fluorimetry. Glycoprotein sequences of escape virus were compared with lyssavirus glycoprotein sequence in the database.Results:In vitro neutralizing activity of the antibody was 691 IU/mg; the neutralization index of the antibody was 8.52 with CVS-11 and 7.05 with CTN. Sequence analysis of the escape mutant virus glycoprotein showed that the key amino acids of the antibody were N37 K, N194 K and K205 N. The amino acid sequence of mutated virus glycoprotein was compared with that of 1 338 lyssavirus glycoprotein. The result showed that only one strain of lyssavirus had the same amino acid type at position 194 as that of mutated escape strain. Tm (melting temperature) of the antibody was 70.58±0.31 ℃.Conclusions:A recombinant human neutralizing antibody with high neutralization index was obtained, which has high thermal stability.
