Sequence analysis of variable regions of human monoclonal anti-P immunoglobulin
10.13303/j.cjbt.issn.1004-549x.2026.01.004
- VernacularTitle:人源单克隆抗-P免疫球蛋白基因可变区序列分析
- Author:
Zhonghui GUO
1
;
Dong XIANG
1
;
Qin LI
1
;
Ziyan ZHU
1
Author Information
1. Shanghai Blood Center, Shanghai 200051, China
- Publication Type:Journal Article
- Keywords:
human monoclonal anti-P antibody;
complementarity determining region (CDR);
germline gene;
gene rearrangement
- From:
Chinese Journal of Blood Transfusion
2026;39(1):24-30
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To identify the structure of the complementarity determining region (CDRs), the V(D)J rearrangement and somatic hypermutational characteristics of the heavy and light chains of a red blood cell blood group-specific monoclonal antibody. Methods: The hybridoma cell line secreting human IgM κ monoclonal anti-P antibody was used as the research object. Total RNA was extracted from cultured monoclonal cell line, and cDNA was obtained by reverse transcription PCR (RT-PCR) using random hexamers primers. It was then amplified and sequenced using primers specific for variable regions of the immunoglobulin heavy and light chains encoding the anti-P antibody. The sequences were aligned against the NCBI database using online Immunoglobulin BLAST (Ig-BLAST) tool. Results: The study determined the structure of the CDRs and framework regions (FRs) of the variable regions of human monoclonal anti-P immunoglobulin, as well as the characteristics of V(D)J rearrangement. Moreover, the closest VH, VD, and VJ germline alleles for the heavy chain and VL and VJ germline alleles for the light chain were also identified. The IgH gene rearrangment pattern of the monoclonal anti-P was IGHV6-1
* 01—IGHD5-18
02—IGHJ4
02 and IgL gene was IGκV1-12
01—IGκJ3
01. Nine base mutations occurred within the germline gene IGHV6-1
01 in variable region of heavy chain, whereas 5 base mutations were found in the germline gene IGκV1-12
01 in variable region of light chain, respectively. Conclusion: This study characterized the CDR structure in monoclonal antibody cell line targeting the high-frequency red blood cell P antigen, and provided a foundation for the construction of recombinant antibody expressing plasmids and transfomation of the immunoglobulin type.
