Production of monoclonal antibodies using a secretion capture report web.
We describe a novel method for the production of monoclonal antibodies using a secretion capture report web (SCRW). Following HAT selection in bulk culture, individual hybridomas are encapsulated in biotinylated agarose drops. Antibody secreted by the hybridoma is captured within the agarose drop using an avidin bridge and biotinylated anti-mouse immunoglobulin. The secreted antibody is detected by a fluorescent reporter which can be either a second anti-mouse antibody or an antigen. The binding of the reporter can be quantitated and the desired hybridoma directly cloned by flow cytometry. Multiparameter (i.e., two-color) reporter analysis can also be used to selectively enrich and clone rare hybridomas secreting antibodies directed to unique epitopes. The method allows the characterization of thousands of clones per second and the isolation of hundreds of clones per day.
Kenney
J S
JS
Department of Leukocyte Biology, Syntex Research, Palo Alto, California 94304, USA. john.kenney@syntex.com
Gray
F
F
Ancel
M H
MH
Dunne
J F
JF
D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D004339:Drug Compounding; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006801:Humans; D006825:Hybridomas / Q000276:immunology* / Q000378:metabolism; D007158:Immunologic Techniques; D011993:Recombinant Fusion Proteins / Q000276:immunology; D012685:Sepharose
D000911:Antibodies, Monoclonal; D011993:Recombinant Fusion Proteins; D012685:Sepharose