title: High throughput production of mouse monoclonal antibodies using antigen microarrays.
abstract: Recent advances in proteomics 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 6-10 wk 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.
authors.affiliation: Biochemical Instrumentation Programme, European Molecular Biology Laboratory, Heidelberg, Germany.
authors.forename: Federico
authors.initials: F
authors.lastname: De Masi
authors.forename: Pieranna
authors.initials: P
authors.lastname: Chiarella
authors.forename: Heike
authors.initials: H
authors.lastname: Wilhelm
authors.forename: Marzia
authors.initials: M
authors.lastname: Massimi
authors.forename: Belinda
authors.initials: B
authors.lastname: Bullard
authors.forename: Wilhelm
authors.initials: W
authors.lastname: Ansorge
authors.forename: Alan
authors.initials: A
authors.lastname: Sawyer
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification*; D000941:Antigens / Q000737:chemistry* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D006825:Hybridomas / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D040081:Protein Array Analysis
chemical_list: D000911:Antibodies, Monoclonal; D000941:Antigens