PUBLICATION
A rapid and scalable method for selecting recombinant mouse monoclonal antibodies
- Authors
- Crosnier, C., Staudt, N., and Wright, G.J.
- ID
- ZDB-PUB-100614-14
- Date
- 2010
- Source
- BMC Biology 8: 76 (Journal)
- Registered Authors
- Staudt, Nicole, Wright, Gavin J.
- Keywords
- none
- MeSH Terms
-
- Animals
- Antibodies, Monoclonal/genetics*
- Antibodies, Monoclonal/immunology
- Antibodies, Monoclonal/isolation & purification
- Cell Line
- Cell Line, Tumor
- Cloning, Molecular/methods*
- Gene Expression
- Humans
- Hybridomas/immunology
- Immunoglobulin G/immunology
- Male
- Mice
- Mice, Inbred BALB C
- Recombinant Proteins/genetics*
- Recombinant Proteins/immunology
- Recombinant Proteins/isolation & purification
- Zebrafish/embryology
- Zebrafish/immunology
- PubMed
- 20525357 Full text @ BMC Biol.
Citation
Crosnier, C., Staudt, N., and Wright, G.J. (2010) A rapid and scalable method for selecting recombinant mouse monoclonal antibodies. BMC Biology. 8:76.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping