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AssigneeGenentech, Inc.
InventorsLisa Garrard, Dennis Henner, Steven Bass, Ronald Greene, Henry Lowman, James Wells, David Matthews
Estimating the term…
A method for selecting novel proteins such as growth hormone and antibody fragment variants having altered binding properties for their respective receptor molecules is provided. The method comprises fusing a gene encoding a protein of interest to the carboxy terminal domain of the gene III coat protein of the filamentous phage M13. The gene fusion is mutated to form a library of structurally related fusion proteins that are expressed in low quantity on the surface of a phagemid particle. Biological selection and screening are employed to identify novel ligands useful as drug candidates. Disclosed are preferred phagemid expression vectors and selected human growth hormone variants.
A method for selecting novel binding polypeptides comprising:
A human growth hormone variant wherein hGH amino acids 172, 174, 176 and 178 respectively are as a group sequentially selected from one of the following:
A phagemid comprising a replicable expression vector comprising a transcription regulatory element operably linked to a gene fusion encoding a fusion protein wherein the gene fusion comprises a first gene encoding a polypeptide, and a second gene encoding at least a portion of a phage coat protein, wherein a DNA triplet codon encoding an mRNA suppressible terminator codon selected from UAG, UAA and UGA is inserted between the fused ends of the first and second genes, or is substituted for an amino acid encoding triplet codon adjacent to the gene fusion junction.
A human growth variant wherein hGH amino acids 10, 14, 18, and 21 respectively are as a group sequentially selected from one of the following:
A human growth variant wherein hGH amino acids 174 is serine and 176 is tyrosine and hGH amino acids 167, 171, 175 and 179 respectively are as a group sequentially selected from one of the following:
A method for selecting novel binding polypeptides comprising
A human growth hormone (hGH) variant wherein hGH amino acid glutamate174 is replaced by serine174 and phenylalanine176 is replaced by tyrosine176 and one or more of the eight naturally occurring hGH amino acids F10, M14, H18, H21, R167, D171, T175 and I179 are replaced by another natural amino acid.
A method for selecting novel binding polypeptides comprising:
Dependencies are inferred from the claim text.