Establishment of human monoclonal hybridoma cell lines specific for PP1Pk antigens from peripheral B lymphocytes of a p[Tj(a-)] donor

Abstract

Objective To establish lymphoblastoid cell line (LCL) and human monoclonal antibody cell lines against PP1Pk antigens from a donor with rare p[Tj(a-)] phenotype. Methods Peripheral blood mononuclear cells (PBMCs)of an O type p[Tj(a-)] donor were prepared and transformed with Epstein-Barr virus (EBV) in vitro. Normal AB and O type red blood cells treated with bromelain as well as p[Tj (a-)] were used to screen the culture supernatant. The transformed LCL secreting antibodies to PP1Pk antigens was picked up and then hybridized with the SHM-D33 cell line using electric fusion technique. The hybridomas were performed by HOTD selective culture, antibody screening and limited dilution subcloning. Results We established a lymphoblastoid cell line 5E1-E5 and several monoclonal hybridoms cell lines secreting anti-PP1Pk immunoglobulin M from EBV transformed B lymphocytes of a p[Tj(a-)] type donor with anti-PP1Pk successfully. Conclusion Our work made it possible for screening rare p[Tj(a-)] blood type in donor population in China using human monoclonal anti-PP1Pk antibodies, which was important for the establishment of rare blood group banks in China.

Keywords

• p[tj(a-)]
• anti-pp1pk
• anti-tja
• hybridomas technique
• human monoclonal antibody