Efficient generation in vitro, from human peripheral blood cells, of monoclonal Epstein-Barr virus transformants producing specific antibody to a variety of antigens without prior deliberate immunization.
This paper describes a simple protocol for the efficient generation of large numbers of human monoclonal antibody-producing cells. This system is based on initial limiting-dilution culture after Epstein-Barr virus exposure of highly enriched precursors selected from peripheral blood mononuclear cells. Precursors can be enriched by using rosetting or panning approaches. Antibodies to erythrocytes, a mouse mammary carcinoma, DNA, and sperm antigens, produced without any deliberate immunization, are described. Large-scale human monoclonal antibody production may be facilitated by a combination of this protocol with a human cellular fusion system. For efficient precursor analysis and short-term (2 months or more) monoclonal antibody production, however, the system described here may be sufficient.
D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology*; D000936:Antigen-Antibody Complex; D000956:Antigens, Viral / Q000276:immunology*; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D002645:Chickens; D004247:DNA / Q000276:immunology; D005455:Fluorescent Antibody Technique; D005838:Genotype; D006386:Hemagglutination Tests; D004854:Herpesvirus 4, Human / Q000276:immunology*; D006801:Humans; D008297:Male; D009000:Monocytes / Q000276:immunology*; D011863:Radioimmunoassay; D012397:Rosette Formation; D012617:Sea Urchins; D013094:Spermatozoa / Q000276:immunology
D000914:Antibodies, Viral; D000936:Antigen-Antibody Complex; D000956:Antigens, Viral; D004247:DNA