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US 5,843,701 AGranted
Systematic polypeptide evolution by reverse translation
AssigneeNeXstar Pharmaceticals, Inc.
InventorsLarry Gold, Craig Tuerk, David Pribnow, Jonathan Drew Smith
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Abstract
A method for preparing polypeptide ligands of target molecules wherein candidate mixtures comprised of ribosome complexes or mRNA.multidot.polypeptide copolymers are partitioned relative to their affinity to the target and amplified to create a new candidate mixture enriched in ribosome complexes or mRNA.multidot.polypeptide copolymers with an affinity to the target.
Claims40
A method for making a polypeptide ligand of a target molecule comprising:
- (a) synthesizing a mRNA mixture comprising translatable and nontranslatable regions, wherein said translatable region comprises randomized and fixed sequence coding regions;
- (b) synthesizing a mixture of mRNA.multidot.polypeptide copolymers, each member comprising an mRNA and a polypeptide encoded by its associated mRNA, wherein a portion of said nontranslatable region of said mRNA and a portion of said polypeptide encoded by said fixed sequence coding region form a binding interaction;
- (c) partitioning the mRNA.multidot.polypeptide copolymers with respect to affinity of the copolymers to a desired target molecule;
- (d) amplifying the mRNA of partitioned copolymers to yield a translatable mRNA mixture;
- and
- (e) synthesizing a polypeptide or polypeptides encoded by the mRNA mixture of step (d).
3 more elements
A mixture of mRNA.multidot.polypeptide copolymers comprising:
- an mRNA comprising nontranslatable portions and translatable portions;
- a polypeptide encoded by said mRNA comprising random and fixed sequence regions, wherein said mRNA and polypeptide are bound together by at least a portion of the nontranslatable portion of said mRNA and at least a portion of the fixed sequence region of said polypeptide.
A method for making a polypeptide ligand of a target molecule comprising:
- (a) synthesizing a mRNA mixture of at least 10.sup.14 sequences comprising translatable and nontranslatable regions;
- (b) synthesizing a mixture of mRNA.multidot.polypeptide copolymers, each member comprising an mRNA and a polypeptide encoded by its associated mRNA, and not containing a ribosome;
- (c) partitioning the mRNA.multidot.polypeptide copolymers with respect to affinity of the copolymers to a desired target molecule;
- (d) amplifying the mRNA of partitioned copolymers to yield a translatable mRNA mixture;
- and
- (e) synthesizing a polypeptide or polypeptides encoded by the mRNA mixture of step (d).
3 more elements
Dependencies are inferred from the claim text.