title: TRAP display: a high-speed selection method for the generation of functional polypeptides.
abstract: Here, we describe a novel method that enables high-speed in vitro selection of functional peptides, peptidomimetics, and proteins via a simple procedure. We first developed a new cell-free translation system, the TRAP system (transcription-translation coupled with association of puromycin linker), which automatically produces a polypeptide library through a series of sequential reactions: transcription, association of puromycin-DNA linker, translation, and conjugation between the nascent polypeptide and puromycin-DNA linker. We then applied the TRAP system for the selection of macrocyclic peptides against human serum albumin. Six rounds of selection using TRAP display were performed in approximately 14 h, yielding macrocyclic peptides with nanomolar affinity to their target protein. Because TRAP display enables high-speed selection of functional polypeptides, it will facilitate the generation of various polypeptides that are useful for biological and therapeutic applications.
authors.affiliation: Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro-ku, Tokyo 153-8902, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Ishizawa
authors.forename: Takashi
authors.initials: T
authors.lastname: Kawakami
authors.forename: Patrick C
authors.initials: PC
authors.lastname: Reid
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Murakami
mesh_terms: D004247:DNA / Q000737:chemistry; D006801:Humans; D008958:Models, Molecular; D019151:Peptide Library; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D011691:Puromycin / Q000737:chemistry*; D012709:Serum Albumin / Q000037:antagonists & inhibitors; D013329:Structure-Activity Relationship
chemical_list: D019151:Peptide Library; D010455:Peptides; D012709:Serum Albumin; D011691:Puromycin; D004247:DNA