Practical Tips for Construction of Custom Peptide Libraries and Affinity Selection by Using Commercially Available Phage Display Cloning Systems

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...

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Main Authors: Keisuke Fukunaga, Masumi Taki
Format: Article
Language:English
Published: Wiley 2012-01-01
Series:Journal of Nucleic Acids
Online Access:http://dx.doi.org/10.1155/2012/295719
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author Keisuke Fukunaga
Masumi Taki
author_facet Keisuke Fukunaga
Masumi Taki
author_sort Keisuke Fukunaga
collection DOAJ
description 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.
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record_format Article
series Journal of Nucleic Acids
spelling doaj-art-504d658e42d84d4aaf9b1fe9be9b29432025-02-03T01:25:54ZengWileyJournal of Nucleic Acids2090-02012090-021X2012-01-01201210.1155/2012/295719295719Practical Tips for Construction of Custom Peptide Libraries and Affinity Selection by Using Commercially Available Phage Display Cloning SystemsKeisuke Fukunaga0Masumi Taki1Bioscience 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, JapanBioscience 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, JapanPhage 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.http://dx.doi.org/10.1155/2012/295719
spellingShingle Keisuke Fukunaga
Masumi Taki
Practical Tips for Construction of Custom Peptide Libraries and Affinity Selection by Using Commercially Available Phage Display Cloning Systems
Journal of Nucleic Acids
title Practical Tips for Construction of Custom Peptide Libraries and Affinity Selection by Using Commercially Available Phage Display Cloning Systems
title_full Practical Tips for Construction of Custom Peptide Libraries and Affinity Selection by Using Commercially Available Phage Display Cloning Systems
title_fullStr Practical Tips for Construction of Custom Peptide Libraries and Affinity Selection by Using Commercially Available Phage Display Cloning Systems
title_full_unstemmed Practical Tips for Construction of Custom Peptide Libraries and Affinity Selection by Using Commercially Available Phage Display Cloning Systems
title_short Practical Tips for Construction of Custom Peptide Libraries and Affinity Selection by Using Commercially Available Phage Display Cloning Systems
title_sort practical tips for construction of custom peptide libraries and affinity selection by using commercially available phage display cloning systems
url http://dx.doi.org/10.1155/2012/295719
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