Monoclonal antibodies isolated by large-scale screening are suitable for labeling adult zebrafish (Danio rerio) tissues and cell structures.
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 tissue and cell 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.
Deflorian
Gianluca
G
Fondazione IFOM (Istituto FIRC di Oncologia Molecolare), Milan, Italy.
Cinquanta
Mario
M
Beretta
Carlo
C
Venuto
Annunziata
A
Santoriello
Cristina
C
Baldessari
Danila
D
Pezzimenti
Federica
F
Aliprandi
Marisa
M
Mione
Marina
M
de Marco
Ario
A
D000367:Age Factors; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D015153:Blotting, Western; D018604:Epitope Mapping; D006825:Hybridomas / Q000378:metabolism; D007114:Immunization; D007143:Immunoglobulin Heavy Chains / Q000235:genetics / Q000302:isolation & purification; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000302:isolation & purification; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D051379:Mice; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence; D019151:Peptide Library; D011994:Recombinant Proteins / Q000302:isolation & purification; D015027:Zebrafish; D029961:Zebrafish Proteins / Q000032:analysis / Q000276:immunology*
D000911:Antibodies, Monoclonal; D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region; D019151:Peptide Library; D011994:Recombinant Proteins; D029961:Zebrafish Proteins