Enhancing phage display of antibody fragments using gill-amber suppression

초록

The effect of utilizing Ex12 helper pliage, a mutant M13K07 helper having two amber codons at the gIII (gill-amber), in combination with Escherichia coli host strains belonging to the supE genotype on improving the phage display of antibody fragments was investigated. Because of an inefficient read-through of the UAG codons, Ex 12 helper phage produced approximately 10% of the intracellular wt pIII in the szrpE host cells compared to M13K07. The phage progenies rescued from the supE XL-I Blue MRF' strain carrying the recombinant phagemid, pCMTG-SP112, by Ex 12 helper phage displayed both antibody-Delta pIII fusion and wt pIII at a ratio of 1: 1.5, and achieved a 50-fold greater display of the antibody-Delta pIII compared to those obtained by a conventional phage rescue using M13K07. Additionally observed were a 100-fold increase in antigen-binding functionality and a drastic improvement on antigen-specific panning efficiency by the phage progenies. Our approach permits the display of at least one antibody fragment as well as more than one copy of wt pIII on the surface of recombinant phages, and this would make the phagemid-based phage display technology more practical and reliable. (c) 2006 Elsevier B.V. All rights reserved.

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