Patent ID: 6613514
Filing Date: 2003-09-02
Classification: A61K,C07K,C12N,C12Q,C40B,G01N

Abstract:
A method of screening or selecting a further evolved polypeptide for a desired property, the method comprising:(a) providing a mixture of nucleic acid subsequences of two or more parental nucleic acid substrate molecules encoding homologous polypeptides, wherein each parental nucleic acid substrate molecule differs from at least one other parental nucleic acid substrate molecule in at least one nucleotide; (b) ligating two or more of the nucleic acid subsequences together in an ordered reassembly to produce a library of recombinant nucleic acid substrate molecules encoding varients of the homologous polypeptides, which library comprises a plurality of nucleic acid molecules, each member of the plurality comprising at least one subsequence from at least two of the two or more parental nucleic acid substrate molecules, wherein the plurality of nucleic acid molecules are homologous to one another and to the parental substrate nucleic acid molecules; (c) expressing the library of recombinant nucleic acid substrate molecules to provide evolved polypeptides; (d) screening or selecting the evolved polypeptides to identify at least one evolved polypeptide with the desired property; (e) recovering at least one recombinant nucleic acid substrate molecule encoding the at least one evolved polypeptide with the desired property; (f) recombining the at least one recombinant nucleic acid substrate molecule of (e) with at least one polynucleotide to produce at least one further evolved nucleic acid substrate molecule; (g) expressing the at least one further evolved nucleic acid substrate molecule to provide at least one further evolved polypeptide; and, (h) screening or selecting the at least one further evolved polypeptide for the desired property.