Application of recombinant phage display antibody system in study of Codakia orbicularis gill proteins
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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.
Index EntriesAntigen capture Codakia orbicularis gill proteins phage antibodies single-chain fragment variable
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- 10.Kontermann, R. and Dubel, S. (2001), Antibody Engineering, Springer Lab Manual, Heidelberg.Google Scholar
- 15.Chae, J.-S., Choi, J.-K., Lim, H.-T., and Cha, S.-H. (2000), Mol. Cell 11, 7–12.Google Scholar
- 24.McCafferty, J. and Johnson, K. S. (1996), in Phage Display of Peptides and Proteins: A Laboratory Manual, Kay, B. K., Winter, J., and McAfferty, J., eds., Academic, San Diego, pp. 227–276.Google Scholar