EVALUATION OF ANTIBODIES FUSED TO MINOR COAT PROTEIN-III AND MAJOR COAT PROTEIN-VIII OF BACTERIOPHAGE M13

A gene coding for an anti-(2-phenyl-5-oxazolone) single-chain Fv antib ody (Ab) fragment (anti-phOx scFv) was cloned in-frame into phagemid v ectors upstream from genes encoding (i) the wild-type (wt) minor coat protein (cp) III of the filamentous bacteriophage M13 of Escherichia c oli, (ii) a truncated version of cpIII (amino acid (aa) positions 198- 406), (iii) the wt major cpVIII, or (iv) a hybrid of interleukin-1 bet a (IL-1 beta; aa 10-152) and wt cpVIII. Recombinant (re-) phage obtain ed by phagemid rescue were examined for the efficiency of displaying t hese various anti-phOx scFv::cp hybrids with commercially available an ti-M13 enzyme-linked immunosorbent assays (ELISA), by immunoblotting w ith an anti-c-myc Ab, and by selection experiments. We found that the highest ELISA signals were obtained with the cpIII constructs and also that more immunoreactive material was detected by blotting than with Ab::cpVIII fusions. Consequently, more scFv::cpIII than scFv::cpVIII p hage could be recovered in micropanning experiments with the antigen p hOx as target.