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PMID: 16254927 Published · ppublish English Journal Article

High throughput production of mouse monoclonal antibodies using antigen microarrays.

Proteomics ·Vol. 5 ·No. 16 ·2005-11-00 ·Pages 4070-81

De Masi F, Chiarella P, Wilhelm H, Massimi M, Bullard B, Ansorge W, Sawyer A

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.

MeSH Terms
Animals Antibodies, Monoclonal/immunology,isolation & purification Antigens/chemistry,immunology Enzyme-Linked Immunosorbent Assay Hybridomas/immunology Mice Mice, Inbred BALB C Protein Array Analysis
Chemicals
Antibodies, Monoclonal Antigens
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
De Masi Federico
Biochemical Instrumentation Programme, European Molecular Biology Laboratory, Heidelberg, Germany.
Chiarella Pieranna
Wilhelm Heike
Massimi Marzia
Bullard Belinda
Ansorge Wilhelm
Sawyer Alan
Article Info
Journal
Proteomics
Abbr.
Proteomics
ISSN
1615-9853
Published
2005-11-00
Pages
4070-81
Language
English
Region
Germany
NLM ID
101092707
Subset
IM
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