Application of recombinant phage display antibody system in study of Codakia orbicularis gill proteins.
We used the recombinant phage display antibody system (RPAS) to obtain chimeric single-chain fragment variable (ScFv) antibodies to gill proteins of the white clam Codakia orbicularis (Linné, 1758). After three rounds of selection on immunotubes loaded with total gill protein extract, recombinant phages exhibiting antibodies to gill proteins were isolated and tested by enzyme-linked immunosorbent assay (ELISA). Clones exhibiting a high affinity for the mollusk proteins were selected for production of soluble ScFv antibodies, which were purified for subsequent analysis. ScFv antibodies exhibited a reaction specific for a protein whose molecular mass was about 15,000 Daltons and that was detected by the antigen capture technique followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis and Western blotting.
Gourdine
Jean-Philippe
JP
Département de Biologie, Equipe DYNECAR EA 926, UFR Sciences Exactes et Naturelles, Université des Antilles et de la Guyane, 97 159 Pointe-à-Pitre, Guadeloupe.
Greenwell
Pamela
P
Smith-Ravin
Emilie
E
D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology / Q000302:isolation & purification; D001435:Bacteriophages / Q000276:immunology*; D049872:Bivalvia / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D015698:Genomic Library; D005880:Gills / Q000737:chemistry*; D007118:Immunoassay / Q000379:methods; D007135:Immunoglobulin Variable Region / Q000276:immunology; D011506:Proteins / Q000032:analysis*; D011995:Recombination, Genetic
D000914:Antibodies, Viral; D007135:Immunoglobulin Variable Region; D011506:Proteins