Isolation and functional identification of HIV-1 envelope specific monoclonal antibodies
10.3760/cma.j.cn112866-20210114-00010
- VernacularTitle:一组靶向HIV膜蛋白的单克隆抗体的分离和功能鉴定
- Author:
Yuanyuan HU
1
;
Jiaxing KANG
;
Yanling HAO
;
Li REN
;
Shuo WANG
;
Kunxue HONG
;
Yiming SHAO
;
Zheng WANG
;
Dan LI
Author Information
1. 中国疾病预防控制中心性病艾滋病预防控制中心 传染病预防控制国家重点实验室，北京 102206
- Keywords:
HIV-1;
neutralizing activity;
epitope;
monoclonal antibody
- From:
Chinese Journal of Experimental and Clinical Virology
2021;35(2):140-146
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To isolate HIV envelope specific monoclonal antibodies (mAbs) from a chronic HIV-1 subtype B′-infected individual with broadly neutralizing activity, and identify their binding activity, epitopes and neutralizing activity.Methods:Single B cell sorting were used to isolate antigen-specific B cells from an HIV-1 infected individual. Variable regions of heavy chain and light chain gene of mAbs were obtained by PCR amplification, and were ligated into the expression vector subsequently. After expression and purification of mAbs, the binding activity and neutralizing ability of mAbs were identified by ELISA and neutralization assay respectively.Results:Seven mAbs were obtained from the HIV-1 infected individual. The heavy chain genes of the mAbs are derived from IGHV1-69*09, IGHV1-08*01, IGHV1-46*01, IGHV4-34*01, IGHV4-61*01 and IGHV3-43*02 germline, SHM rates range from 10.76% to 21.05%. The light chain genes of the mAbs originate from IGKV3-20*01, IGKV1-39*01, IGKV1-NL1*01, IGKV3-15*01 and IGKV1-9*01 germline, SHM rates cover between 6.03% and 18.64%. Gp140 was well recognized by all seven antibodies. 508-B1, 508-C1, 508-C5, 508-D4 and 508-E5 are gp41-directed mAbs. 508-D6 and 508-F1 bind with gp120. 508-D4 and 508-F1 could neutralize pseudoviruses CNE8 and X2278 in Global Panel with IC 50 values of 38.1μg/ml and 41.7 μg/ml, respectively. Conclusions:This study successfully identified seven envelope specific mAbs, among them five mAbs are directed to gp41 region and two mAbs are targeted to gp120. Two mAbs showed limited neutralization spectrum. These mAbs may provide technical reserves for developing HIV-1 diagnostics kit, antibody-related drugs and vaccines.
