|ZFIN ID: ZDB-PUB-100614-14|
A rapid and scalable method for selecting recombinant mouse monoclonal antibodies
Crosnier, C., Staudt, N., and Wright, G.J.
|Source:||BMC Biology 8: 76 (Journal)|
|Registered Authors:||Staudt, Nicole, Wright, Gavin J.|
|PubMed:||20525357 Full text @ BMC Biol.|
Crosnier, C., Staudt, N., and Wright, G.J. (2010) A rapid and scalable method for selecting recombinant mouse monoclonal antibodies. BMC Biology. 8:76.
ABSTRACTBACKGROUND: 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.