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Article  February 1, 2015

Introduction of Plasmid Encoding for Rare tRNAs Reduces Amplification Bias in Phage Display Biopanning

SRI Authors Michael McGuire, Kathlynn Brown

Citation

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Umlauf, B. J., McGuire, M. J., & Brown, K. C. (2015). Introduction of plasmid encoding for rare tRNAs reduces amplification bias in phage display biopanning. Biotechniques, 58(2), 81-84. doi: 10.2144/000114256

Abstract

Amplification bias is a major hurdle in phage display protocols because it imparts additional, unintended selection pressure beyond binding to the desired target. One potential source of amplification bias is the inherent lack of codon optimization that occurs within phage display libraries. Here we present a method that reduces amplification bias by addition of a plasmid that encodes six low abundance tRNAs into K91 Escherichia coli. This new strain, termed K91+, is used to amplify phage during the selection process. We demonstrate the importance of rare codon usage in phage production, and our method produced an overall increase in uniformity of phage production in a random library. Both of these variables are improved in E. coli K91+ compared with the parental K91 strain. This simple solution, requiring only a commercially available plasmid and an additional antibiotic, can reduce amplification bias in phage display protocols.

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