A new approach to the generation of human or murine antibody producing hybridomas.
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.
Steenbakkers
P G
PG
Department of Immunology, Organon International B.V., Oss, Netherlands.
van Meel
F C
FC
Olijve
W
W
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D000914:Antibodies, Viral / Q000096:biosynthesis; D000921:Antibody-Producing Cells / Q000378:metabolism*; D002459:Cell Fusion; D002460:Cell Line; D002461:Cell Line, Transformed; D015483:HIV Antibodies / Q000096:biosynthesis; D006801:Humans; D006825:Hybridomas / Q000378:metabolism*; D007158:Immunologic Techniques; D008213:Lymphocyte Activation; D051379:Mice; D013154:Spleen; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D015483:HIV Antibodies; C040901:rubella antibodies