Patent ID: 7947475
Filing Date: 2011-05-24
Classification: A61K,A61Q,C07K

Abstract:
1. A method for producing a modified variant BBPI in a bacterial cell, said method comprising: a) mutating a polynucleotide that encodes an unmodified precursor variant BBPI to generate a polynucleotide construct encoding a modified variant BBPI, wherein the unmodified precursor variant BBPI comprises a BBPI scaffold selected from the group consisting of: BBI (SEQ ID NO: 13), BBIt (SEQ ID NO:185), BBI-AV (SEQ ID NO:186), BBIt-AV (SEQ ID NO:187), BBIt-VEGK (SEQ ID NO:640), BBIt-VEGT (SEQ ID NO:641), BBIt-VEGKD (SEQ ID NO:642), BBdb (SEQ ID NO:449), BBsb3 (SEQ ID NO:450), BBtc (SEQ ID NO:451), BBdb-AV (SEQ ID NO:452), BBsb3-AV (SEQ ID NO:453) and BBtc-AV (SEQ ID NO:454), wherein the mutation in step (a) results in an amino acid substitution in the backbone of the unmodified precursor variant at least at one amino acid position chosen from positions equivalent to 1, 4, 5, 11, 13, 18, 25, 27, 29, 31, 38, 40, 50, 52, 55, and 65 of SEQ ID NO: 187, wherein the modified variant BBPI comprises the unmodified precursor variant BBPI with said amino acid substitution in the backbone; and b) introducing said polynucleotide construct encoding a modified variant BBPI into a bacterial host cell; c) culturing said bacterial cell under suitable culture conditions to allow expression of said modified variant BBPI; and d) producing said modified variant BBPI, wherein said modified variant BBPI has a greater trypsin inhibitory activity than the corresponding unmodified precursor variant BBPI.