Efficient display of scFv antibodies on bacteriophage lambda.
In the present work we demonstrate the efficient display of functional scFv antibodies on the bacteriophage lambda capsid. A single-chain (scFv) anti-CEA antibody gene was cloned in two different vectors to obtain fusion of the scFv antibody to the N- or C-terminus of the bacteriophage lambda capsid protein D (gpD). Lambda bacteriophage assembly occurs in the reducing environment of the cytoplasm; despite this the lambda-displayed anti-CEA antibody fragments retain the capacity to recognize the antigen, indicating correct single-chain antibody folding. Efficient production of functional scFv exposed on lambda capsid with viable antigen binding specificity allowed us to study and compare the capacity of display, the stability of recombinant antibody expression and the assembly efficiency of bacteriophage particles decorated with recombinant antibody fused to the amino- or carboxy-terminus of lambda D protein.
Vaccaro
Paola
P
Kenton Labs, c/o Sigma-Tau, Pomezia (Rome), Italy.
Pavoni
Emiliano
E
Monteriù
Giorgia
G
Andrea
Pucci
P
Felici
Franco
F
Minenkova
Olga
O
D010582:Bacteriophage lambda / Q000276:immunology*; D015153:Blotting, Western; D036022:Capsid Proteins / Q000276:immunology; D002272:Carcinoembryonic Antigen / Q000276:immunology; D002846:Chromatography, Affinity; D006801:Humans; D007128:Immunoglobulin Fragments / Q000276:immunology*; D007135:Immunoglobulin Variable Region / Q000276:immunology; D009842:Oligopeptides; D019151:Peptide Library; D010455:Peptides / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology
D036022:Capsid Proteins; D002272:Carcinoembryonic Antigen; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D009842:Oligopeptides; D019151:Peptide Library; D010455:Peptides; D011994:Recombinant Proteins; C055966:immunoglobulin Fv; C085706:FLAG peptide