León Graciela, Cruz Carlos, Rojas Lola
Banco Municipal de Sangre del Distrito Capital, Esquina de Pirineos, San José, Caracas, Venezuela.
Invest Clin. 2004 Jun;45(2):145-57.
The purpose of this research was to produce murine monoclonal antibodies with anti B specificity to be used as reagents in red cells typing. Balb/c mice were immunized with substance B from saliva of a donor secreting B antigen, in order to obtain spleen cells or splenocytes producing antibodies with anti B specificity. These cells were fused with mouse mycloma cells P3-X63.Ag8.653 by using the PEG technique. After seeded, the hybridomas secreting the putative antibodies were cloned through the limiting dilution technique to obtain monoclonality. Afterwards, the antibodies were evaluated using immunological and immunochemical methods. BMS-4 clone secreting antibodies IgM-kappa with anti B specificity were obtained, which displayed high potency, excellent avidity, great sensibility and long time stability. We think that this clone can be used to produce a red cell typing reagent.
本研究的目的是制备具有抗B特异性的鼠单克隆抗体,用作红细胞分型试剂。用分泌B抗原供体的唾液中的物质B免疫Balb/c小鼠,以获得产生具有抗B特异性抗体的脾细胞或脾淋巴细胞。通过聚乙二醇(PEG)技术将这些细胞与小鼠骨髓瘤细胞P3-X63.Ag8.653融合。接种后,通过有限稀释技术对分泌假定抗体的杂交瘤进行克隆以获得单克隆性。之后,使用免疫学和免疫化学方法对抗体进行评估。获得了分泌具有抗B特异性的IgM-κ抗体的BMS-4克隆,其显示出高效能、优异的亲和力、高灵敏度和长期稳定性。我们认为该克隆可用于生产红细胞分型试剂。