ZFIN ID: ZDB-PUB-090505-19
Monoclonal antibodies isolated by large-scale screening are suitable for labeling adult zebrafish (Danio rerio) tissues and cell structures
Deflorian, G., Cinquanta, M., Beretta, C., Venuto, A., Santoriello, C., Baldessari, D., Pezzimenti, F., Aliprandi, M., Mione, M., and de Marco, A.
Date: 2009
Source: Journal of immunological methods   346(1-2): 9-17 (Journal)
Registered Authors: Baldessari, Danila, Deflorian, Gianluca, Mione, Marina, Pezzimenti, Federica, Santoriello, Cristina
Keywords: tissue biopanning, hybridoma cells, immunohistochemistry, recombinant antibodies, zebrafish histology
MeSH Terms:
  • Age Factors
  • Animals
  • Antibodies, Monoclonal/biosynthesis
  • Antibodies, Monoclonal/genetics
  • Antibodies, Monoclonal/isolation & purification*
  • Blotting, Western
  • Epitope Mapping
  • Hybridomas/metabolism
  • Immunization*
  • Immunoglobulin Heavy Chains/genetics
  • Immunoglobulin Heavy Chains/isolation & purification
  • Immunoglobulin Variable Region/genetics
  • Immunoglobulin Variable Region/isolation & purification
  • Immunohistochemistry
  • Immunoprecipitation
  • Mice
  • Mice, Inbred BALB C
  • Microscopy, Fluorescence
  • Peptide Library*
  • Recombinant Proteins/isolation & purification
  • Zebrafish
  • Zebrafish Proteins/analysis
  • Zebrafish Proteins/immunology*
PubMed: 19410577 Full text @ J. Immunol. Methods
The lack of a sufficient number of antibodies represents an obstacle in the research performed using the zebrafish (Danio rerio) as a model organism. On the other hand, high throughput generation of antibodies, especially those suitable for immunohistochemistry, is not an established methodology. Here we present the results of an immunization experiment with a zebrafish tissue lysate that allowed for the isolation of hundreds of monoclonal antibodies suitable for labeling of a large variety of zebrafish tissues and cells structures. Some of them were further characterized in terms of detailed localization and age-dependent expression. In addition, the antigen recognized by one of them was first immunoprecipitated and then identified by mass spectrometry. Furthermore, immunofluorescence-competent recombinant antibodies were also isolated by panning large repertoire phage display libraries, in both single chain (scFv) and single domain (VHH) format. Such selection alternative is simpler to organize and could contribute to limit the costs of antibody screening and production.