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Proceedings of the National Academy of Sciences of the United States of AmericaVolume 94, Issue 10, 13 May 1997, Pages 4937-4942

In vitro selection and evolution of functional proteins by using ribosome display(Article)

  • Biochemisches Institut, Universität Zürich, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland

Abstract

We report here a system with which a correctly folded complete protein and its encoding mRNA both remain attached to the ribosome and can be enriched for the ligand-binding properties of the native protein. We have selected a single-chain fragment (scFv) of an antibody 108-fold by live cycles of transcription, translation, antigen-affinity selection, and PCR. The selected scFv fragments all mutated in vitro by acquiring up to four unrelated amino acid exchanges over the five generations, but they remained fully compatible with antigen binding. Libraries of native folded proteins can now be screened and made to evolve in a cell-free system without any transformation or constraints imposed by the host cell.

Indexed keywords

EMTREE medical terms:articlecell free systemgenetic engineeringligand bindingpolymerase chain reactionpriority journalprotein foldingribosome
MeSH:Amino Acid SequenceBase SequenceBinding SitesDNA PrimersEvolutionImmunoglobulin FragmentsInformation SystemsMolecular Sequence DataMutagenesis, Site-DirectedPoint MutationPolymerase Chain ReactionProtein BiosynthesisProtein EngineeringProtein FoldingRecombinant ProteinsRibosomesRNA, MessengerTranscription, Genetic

Chemicals and CAS Registry Numbers:

DNA Primers; Immunoglobulin Fragments; Recombinant Proteins; RNA, Messenger

  • ISSN: 00278424
  • CODEN: PNASA
  • Source Type: Journal
  • Original language: English
  • DOI: 10.1073/pnas.94.10.4937
  • PubMed ID: 9144168
  • Document Type: Article

  Pluckthun, A.; Biochemisches Institut, Universitat Zurich, Winterthurerstrasse 190, Switzerland;
© Copyright 2007 Elsevier B.V., All rights reserved.

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