High throughput production of mouse monoclonal antibodies using antigen microarrays

被引:64
作者
De Masi, F
Chiarella, P
Wilhelm, H
Massimi, M
Belinda, B
Ansorge, W
Sawyer, A
机构
[1] European Mol Biol Lab, Monoclonal Antibody Core Facil, Mouse Biol Programme, I-00016 Monterotondo, Italy
[2] European Mol Biol Lab, Biochem Instrumentat Programme, Heidelberg, Germany
[3] European Mol Biol Lab, Struct Biol Programme, Heidelberg, Germany
[4] DKFZ, Heidelberg, Germany
关键词
hybridoma production; microarrays; monoclonal antibodies; multiplexing;
D O I
10.1002/pmic.200401279
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:4070 / 4081
页数:12
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