In vitroselection and evolution of functional proteins by using ribosome display
0301 basic medicine
Protein Folding
Binding Sites
Base Sequence
Transcription, Genetic
Molecular Sequence Data
Protein Engineering
Biological Evolution
Polymerase Chain Reaction
Recombinant Proteins
03 medical and health sciences
Protein Biosynthesis
Mutagenesis, Site-Directed
Point Mutation
Amino Acid Sequence
RNA, Messenger
Immunoglobulin Fragments
Ribosomes
DNA Primers
Information Systems
DOI:
10.1073/pnas.94.10.4937
Publication Date:
2002-07-26T14:31:39Z
AUTHORS (2)
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 five cycles of transcription, translation, antigen-affinity selection, and PCR. The selected scFv fragments all mutatedin vitroby 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.
SUPPLEMENTAL MATERIAL
Coming soon ....
REFERENCES (41)
CITATIONS (875)
EXTERNAL LINKS
PlumX Metrics
RECOMMENDATIONS
FAIR ASSESSMENT
Coming soon ....
JUPYTER LAB
Coming soon ....