Open AccessIntegration of phage and yeast display platforms: A reliable and cost effective approach for binning of peptides as displayed on-phage
Author(s) -
Priyanka Pandya,
Robert O. Sayers,
Joey Ting,
Shaghayegh Morshedian,
Carina Torres,
Justine S. Cudal,
Kai Zhang,
Jonathan R. Fitchett,
Qing Zhang,
Feiyu F. Zhang,
Jing Wang,
Jim D. Durbin,
Juan J. Carrillo,
Alfonso Espada,
Howard B. Broughton,
Yuewei Qian,
Sepideh Afshar
Publication year - 2020
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0233961
Subject(s) - panning (audio) , phage display , yeast , peptide library , peptide , computational biology , population , chemistry , biology , biochemistry , peptide sequence , gene , paleontology , zoom , demography , sociology , lens (geology)
Hundreds of target specific peptides are routinely discovered by peptide display platforms. However, due to the high cost of peptide synthesis only a limited number of peptides are chemically made for further analysis. Here we describe an accurate and cost effective method to bin peptides on-phage based on binding region(s), without any requirement for peptide or protein synthesis. This approach, which integrates phage and yeast display platforms, requires display of target and its alanine variants on yeast. Flow cytometry was used to detect binding of peptides on-phage to the target on yeast. Once hits were identified, they were synthesized to confirm their binding region(s) by HDX (Hydrogen deuterium exchange) and crystallography. Moreover, we have successfully shown that this approach can be implemented as part of a panning process to deplete non-functional peptides. This technique can be applied to any target that can be successfully displayed on yeast; it narrows down the number of peptides requiring synthesis; and its utilization during selection results in enrichment of peptide population against defined binding regions on the target.