A novel phage display vector for selection of target-specific peptides

Author:

Chang Alex1,Ting Joey P2,Espada Alfonso3,Broughton Howard3,Molina-Martin Manuel3,Afshar Sepideh2

Affiliation:

1. Department of Pharmacy, Santa Clara Valley Medical Center, San Jose CA 95128, USA

2. Protein Engineering, Eli Lilly Biotechnology Center, San Diego, CA 92121, USA

3. Department of Discovery Chemistry Research & Technology, Centro de Investigacion Lilly, Av. de la Industria, 30, 28108 Alcobendas, Madrid, Spain

Abstract

Abstract Intrinsic low display level of polypeptides on phage is a fundamental and limiting hurdle in successful isolation of target-specific binders by phage display technology. To circumvent this challenge, we optimized the copy number of peptides displayed on the phage surface using type 33 phage vector. We randomized the first 67 amino acids of the wild type PIII to identify mutants that would result in its reduced expression. Consequently, the display level was improved by 30-fold due to higher incorporation of the synthetic PIII–peptide fusion protein on the phage surface. Utilization of this novel phage vector should provide a solid basis for the discovery of therapeutic peptides.

Funder

Eli Lilly & Company

Publisher

Oxford University Press (OUP)

Subject

Molecular Biology,Biochemistry,Bioengineering,Biotechnology

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