High throughput production of mouse monoclonal antibodies using antigen microarrays.
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.
De Masi
Federico
F
Biochemical Instrumentation Programme, European Molecular Biology Laboratory, Heidelberg, Germany.
Chiarella
Pieranna
P
Wilhelm
Heike
H
Massimi
Marzia
M
Bullard
Belinda
B
Ansorge
Wilhelm
W
Sawyer
Alan
A
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
D000911:Antibodies, Monoclonal; D000941:Antigens