Preparation of human monoclonal anti-C cell line from peripheral blood B lymphocytes of D--donor
10.13303/j.cjbt.issn.1004-549x.2022.04.011
- VernacularTitle:利用D--外周血B细胞构建单克隆抗-C杂交瘤细胞株
- Author:
Zhonghui GUO
1
;
Fengyong ZHAO
1
;
Demei ZHANG
2
;
Dong XIANG
1
;
Jiamin ZHANG
1
;
Ying YANG
1
;
Qin LI
1
;
Qixiu YANG
1
;
Chen WANG
1
;
Ziyan ZHU
1
Author Information
1. Shanghai Blood Center, Shanghai 200051, China
2. Taiyuan Blood Center
- Publication Type:Journal Article
- Keywords:
D--phenotype;
human monoclonal antibody;
anti-C;
hybridoma technique
- From:
Chinese Journal of Blood Transfusion
2022;35(4):400-404
- CountryChina
- Language:Chinese
-
Abstract:
【Objective】 To establish human hybridoma cell lines, secreting monoclonal antibody against antigens of Rh blood system, from a donor with rare D--phenotype. 【Methods】 Peripheral blood B lymphocytes of an O type female donor, lacking C/c/E/e antigens on her erythrocyte, were transformed with Epstein-Barr virus (EBVs). EBVs were harvested from the cultural supernatant of B95-8 cells. The transformed lymphoblastoid cell line (LCL) secreting antibodies to C antigens were picked up and then hybridized with the myeloma SHM-D33 using electric fusion technique. Hybridoma cells were selected by HATD-Ouabain(HOTD)(Hypoxantine, Aminopterin, Thymidine, 2-Deoxycytide, and Ouabain)culture medium, microplate antibody screening and limited dilution subcloning. The monoclonal antibody was assayed by serological test and was confirmed by flow cytometry (FCM). 【Results】 From the cultural supernatant of D--peripheral blood transformed B lymphocytes, 3A6-C6, which agglutinated with R1R1(DCe/DCe)O-type RBCs but not with R2R2(DcE/DcE)O-type RBCs, was screened and preliminarily identified as anti-C. We established a hybridoma cell line secreting anti-C immunoglobulin M from B cells of D--individual successfully after hybridization with SHM-D33 myeloma cells. 【Conclusion】 The study had laid the groundwork for future research and development of human monoclonal antibodies against Rh antigens of RBC in future for diagnosis and screening purpose.
