Cloning of porcine scFv antibodies by phage display and expression in Escherichia coli.

Veterinary immunology and immunopathology

PubMedID: 14700536

Li F, Aitken R. Cloning of porcine scFv antibodies by phage display and expression in Escherichia coli. Vet Immunol Immunopathol. 2004;97(1-2):39-51.
Oligonucleotide primers were designed for recovery of Ig H, kappa and lambda transcripts from porcine splenic cDNA. The products were cloned and scFvs constructed in a phage display vector. E. coli HB2151 was transformed with the constructs and upon induction, scFvs of the predicted molecular weight could be detected in culture supernatants by Western blotting and immunoprecipitation with anti-pig IgG. Bacteriophage displaying a swine scFv were diluted into an excess of phage carrying a human anti-thyroglobulin scFv and then panned on plastic coated with anti-pig IgG. Rapid enrichment of phage carrying the porcine scFv through three rounds of selection demonstrated the successful display of an authentically folded pig Ig at the viral surface. The data demonstrate that phage display techniques can be applied successfully to porcine Igs and that expression of recombinant pig scFvs in bacteria can be achieved. These techniques offer the opportunity to generate authentic porcine monoclonal reagents for basic and applied studies.