Journal
PROTEOMICS
Volume 5, Issue 16, Pages 4070-4081Publisher
WILEY
DOI: 10.1002/pmic.200401279
Keywords
hybridoma production; microarrays; monoclonal antibodies; multiplexing
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Recent advances in proteornics research underscore the increasing need for high-affinity monoclonal antibodies, which are still generated with lengthy, low-throughput antibody production techniques. Here we present a semi-automated, high-throughput method of hybridoma generation and identification. Monoclonal antibodies were raised to different targets in single batch runs of 6-10wk using multiplexed immunisations, automated fusion and cell-culture, and a novel antigen-coated microarray-screening assay. In a large-scale experiment, where eight mice were immunized with ten antigens each, we generated monoclonal antibodies against 68 of the targets (85%), within 6 wk of the primary immunization.
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