In vitro selection and evolution of functional proteins by using ribosome display.
basic_science · Level V
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- Record sourced from PubMed, PMID 9144168.
- Also identified by PMC identifier 24609.
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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 10(8)-fold by five 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.
Medical subject headings
- Immunoglobulin Fragments
- Protein Biosynthesis
- RNA, Messenger
- Ribosomes