Single B cell sorting-based amplification and functional identification of HIV-1 monoclonal antibody genes

Bin JU; Yanling Hao; Dan LI; Li Ren; Hua Liang; Shuo Wang; Jiali Hou; Min Wei; Yiming Shao

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Single B cell sorting-based amplification and functional identification of HIV-1 monoclonal antibody genes

VernacularTitle: 利用单个B细胞分选方法获得HIV-1单克隆抗体基因及功能鉴定
Author: Bin JU ^{1
,

2} ; Yanling HAO ³ ; Dan LI ³ ; Li REN ³ ; Hua LIANG ³ ; Shuo WANG ³ ; Jiali HOU ³ ; Min WEI ¹ ; Yiming SHAO ^{1
,

2}
Author Information

1. School of Medicine, Nankai University, Tianjin 300071, China
2. State Key Laboratory of Infectious Disease Prevention and Control, National Center for AIDS/STD Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
3. State Key Laboratory of Infectious Disease Prevention and Control, National Center for AIDS/STD Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
Publication Type:Journal Article
Keywords: Single B cell sorting; Antibody gene; Sequence analysis; Binding ability; Neutralizing activity
From: Chinese Journal of Microbiology and Immunology 2018;38(7):481-487
CountryChina
Language:Chinese
Abstract: Objective:To amplify and identify monoclonal antibody genes from HIV-1-infected patients.
Methods:Single cell sorting was used to isolate antigen-specific single B cells. Sequence Identity Matrix and the international ImMunoGeneTics information system were used to analyze antibody variable region genes. Binding abilities were detected by enzyme linked immunosorbent assay. Neutralizing activities were tested by TZM-bl/pseudovirus assay.
Results:The heavy and light chain genes of four, seven, and eleven antibodies were amplified and sequenced from three HIV-1-infected patients, respectively. They were derived from various germline genes with flexible CDR3 lengths and somatic mutations. A1 and B3 antibodies bound to HIV-1 clade B, CRF01_AE, and CRF07_BC antigens. The half maximal inhibitory concentration values of A1 and B3 against MW965 virus were 0.04 μg/ml and 37.34 μg/ml.
Conclusion:In this study, we acquired a lot of monoclonal antibody genes and two HIV-1 monoclonal binding and neutralizing antibodies, which would provide basic data for further research on monoclonal antibody identification.