Practical tips for construction of custom Peptide libraries and affinity selection by using commercially available phage display cloning systems
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Updated: May 18, 2026
A Protocol for Phage Display and Affinity Selection Using Recombinant Protein Baits
Published on: February 16, 2014
Keisuke Fukunaga1, Masumi Taki
1Bioscience and Technology Program, Department of Engineering Science, The Graduate School of Informatics and Engineering, The University of Electro-Communications (UEC), 7-5-1 Chofugaoka, Chofu, Tokyo 182-8585, Japan.
Abstract:
Phage display technology is undoubtedly a powerful tool for affinity selection of target-specific peptide. Commercially available premade phage libraries allow us to take screening in the easiest way. On the other hand, construction of a custom phage library seems to be inaccessible, because several practical tips are absent in instructions. This paper focuses on what should be born in mind for beginners using commercially available cloning kits (Ph.D. with type 3 vector and T7Select systems for M13 and T7 phage, respectively). In the M13 system, Pro or a basic amino acid (especially, Arg) should be avoided at the N-terminus of peptide fused to gp3. In both systems, peptides containing odd number(s) of Cys should be designed with caution. Also, DNA sequencing of a constructed library before biopanning is highly recommended for finding unexpected bias.
