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.
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.
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.