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

被引:8
作者
Chang, Alex [1 ]
Ting, Joey P. [2 ]
Espada, Alfonso [3 ]
Broughton, Howard [3 ]
Molina-Martin, Manuel [3 ]
Afshar, Sepideh [2 ]
机构
[1] Santa Clara Valley Med Ctr, Dept Pharm, San Jose, CA 95128 USA
[2] Eli Lilly Biotechnol Ctr, Prot Engn, San Diego, CA 92121 USA
[3] Ctr Invest Lilly, Dept Discovery Chem Res & Technol, Av Ind 30, Madrid 28108, Spain
关键词
HDX-MS; peptide; phage display; type 33 phage vector; PROLINE; SITE; FRAGMENTS; PROTEINS; PIII; FD;
D O I
10.1093/protein/gzaa023
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 30fold 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.
引用
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页数:8
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