Peptide screening using pure ribosome display

Hiroyuki Ohashi, Takashi Kanamori, Eriko Osada, Bintang K. Akbar, Takuya Ueda

Research output: Chapter in Book/Report/Conference proceedingChapter

8 Citations (Scopus)

Abstract

To demonstrate directed protein evolution or selection of functional polypeptides, ribosome display is one of the most ideal technologies of evolutionary engineering. Intrinsic components, such as nucleases in the cell extract-based cell-free protein synthesis systems, reduce the stability of the messenger RNA-ribosome-polypeptide ternary complex, thereby preventing the attainment of reliable results. To overcome this problem, we have developed an effective and highly controllable ribosome display system using the protein synthesizing using recombinant elements (PURE) system. Since the activities of nucleases and other inhibitory factors are very low in the PURE system, the ternary complex is highly stable and the selected mRNA can be reliably recovered. Using this system, we were able to select peptides that specifically bind to monoclonal antibodies from random peptide libraries. The advantages of the modified PURE system for ribosome display strongly substantiate its usability.

Original languageEnglish
Title of host publicationRibosome Display and Related Technologies
Subtitle of host publicationMethods and Protocols
EditorsJulie Douthwaite, Ronald Jackson
Pages251-259
Number of pages9
DOIs
Publication statusPublished - 2012 Jan 2
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume805
ISSN (Print)1064-3745

Fingerprint

Ribosomes
Peptides
Proteins
Peptide Library
Messenger RNA
Cell Extracts
Monoclonal Antibodies
Technology

Keywords

  • Cell-free protein synthesis system
  • Epitope mapping
  • In vitro selection
  • Peptide screening
  • Protein engineering
  • PURE system
  • Ribosome display

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Ohashi, H., Kanamori, T., Osada, E., Akbar, B. K., & Ueda, T. (2012). Peptide screening using pure ribosome display. In J. Douthwaite, & R. Jackson (Eds.), Ribosome Display and Related Technologies: Methods and Protocols (pp. 251-259). (Methods in Molecular Biology; Vol. 805). https://doi.org/10.1007/978-1-61779-379-0_14

Peptide screening using pure ribosome display. / Ohashi, Hiroyuki; Kanamori, Takashi; Osada, Eriko; Akbar, Bintang K.; Ueda, Takuya.

Ribosome Display and Related Technologies: Methods and Protocols. ed. / Julie Douthwaite; Ronald Jackson. 2012. p. 251-259 (Methods in Molecular Biology; Vol. 805).

Research output: Chapter in Book/Report/Conference proceedingChapter

Ohashi, H, Kanamori, T, Osada, E, Akbar, BK & Ueda, T 2012, Peptide screening using pure ribosome display. in J Douthwaite & R Jackson (eds), Ribosome Display and Related Technologies: Methods and Protocols. Methods in Molecular Biology, vol. 805, pp. 251-259. https://doi.org/10.1007/978-1-61779-379-0_14
Ohashi H, Kanamori T, Osada E, Akbar BK, Ueda T. Peptide screening using pure ribosome display. In Douthwaite J, Jackson R, editors, Ribosome Display and Related Technologies: Methods and Protocols. 2012. p. 251-259. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-61779-379-0_14
Ohashi, Hiroyuki ; Kanamori, Takashi ; Osada, Eriko ; Akbar, Bintang K. ; Ueda, Takuya. / Peptide screening using pure ribosome display. Ribosome Display and Related Technologies: Methods and Protocols. editor / Julie Douthwaite ; Ronald Jackson. 2012. pp. 251-259 (Methods in Molecular Biology).
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