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Production
of Monoclonal Antibodies Using a Secretion Capture
Report Web
Biotechnology, August 1995, vol. 13,
pp.787-790
John S. Kenney, Forest Gray, Marie-Helene
Ancel and John F. Dunne
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.
Multi-parameter (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.
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