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BMC Biol. 2010 Jun 4;8:76. doi: 10.1186/1741-7007-8-76.

A rapid and scalable method for selecting recombinant mouse monoclonal antibodies.

Author information

1
Cell Surface Signalling Laboratory, Wellcome Trust Sanger Institute, Cambridge CB10 1HH, UK.

Abstract

BACKGROUND:

Monoclonal antibodies with high affinity and selectivity that work on wholemount fixed tissues are valuable reagents to the cell and developmental biologist, and yet isolating them remains a long and unpredictable process. Here we report a rapid and scalable method to select and express recombinant mouse monoclonal antibodies that are essentially equivalent to those secreted by parental IgG-isotype hybridomas.

RESULTS:

Increased throughput was achieved by immunizing mice with pools of antigens and cloning - from small numbers of hybridoma cells - the functionally rearranged light and heavy chains into a single expression plasmid. By immunizing with the ectodomains of zebrafish cell surface receptor proteins expressed in mammalian cells and screening for formalin-resistant epitopes, we selected antibodies that gave expected staining patterns on wholemount fixed zebrafish embryos.

CONCLUSIONS:

This method can be used to quickly select several high quality monoclonal antibodies from a single immunized mouse and facilitates their distribution using plasmids.

PMID:
20525357
PMCID:
PMC2898661
DOI:
10.1186/1741-7007-8-76
[Indexed for MEDLINE]
Free PMC Article

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