Steenbakkers P G, van Meel F C, Olijve W
Department of Immunology, Organon International B.V., Oss, Netherlands.
J Immunol Methods. 1992 Jul 31;152(1):69-77. doi: 10.1016/0022-1759(92)90090-g.
A new and very efficient method for the generation of human and murine monoclonal antibodies has been developed. The method is based on clonal expansion of single B cells in the presence of human T cell supernatant and irradiated murine thymoma helper cells. Subsequently, the clonally expanded B cells are immortalized by electric field mediated cell fusion. The high efficiency of the method permits the processing of small numbers of lymphocytes, e.g., obtained by preselection of specific B cells, small amounts of human donor material and murine PBL or lymph node cells. The method may be an alternative for the EBV-transformation technique used for the generation of human monoclonal antibodies, which immortalizes only a subset of B cells and frequently yields poorly growing or unstable cell lines. This report describes the generation of murine anti-HIV and human anti-rubella antibodies combining the clonal expansion of B cells and mini-electrofusion.
一种用于产生人源和鼠源单克隆抗体的全新且高效的方法已被开发出来。该方法基于在人T细胞上清液和经辐照的鼠胸腺瘤辅助细胞存在的情况下单个B细胞的克隆扩增。随后，通过电场介导的细胞融合使克隆扩增的B细胞永生化。该方法的高效性允许处理少量淋巴细胞，例如通过预选特定B细胞、少量人供体材料以及鼠外周血淋巴细胞（PBL）或淋巴结细胞获得的淋巴细胞。该方法可能是用于产生人单克隆抗体的EBV转化技术的一种替代方法，EBV转化技术仅使一部分B细胞永生化，并且经常产生生长不良或不稳定的细胞系。本报告描述了结合B细胞克隆扩增和微量电融合产生鼠抗HIV抗体和人抗风疹抗体的过程。
