Patent · US Active

Methods of synthesizing heteromultimeric polypeptides in yeast using a haploid mating strategy

US7927863B2 · kind B2 · utility

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5References
28Claims
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Key dates

Filing dateMay 8, 2006
Grant dateApr 19, 2011
Priority date
Expiry dateMay 27, 2029

Classification

  • Technology area (CPC C)Chemistry; Metallurgy
  • CPC primaryC07K2317/24
  • WIPO fieldBiotechnology
  • WIPO sectorChemistry

Abstract

Methods are provided for the synthesis and secretion of recombinant proteins preferably large mammalian proteins or hetero-multimeric proteins at high levels and for prolonged time in polyploid, preferably diploid yeast. These methods use various mating competent yeast, including Pichia. In a preferred embodiment, a first expression vector is transformed into a first haploid cell; and a second expression vector is transformed into a second haploid cell. The transformed haploid cells, each individually synthesizing a non-identical polypeptide, are identified and then genetically crossed or fused. The resulting diploid strains are utilized to produce and secrete fully assembled and biologically functional hetero-multimeric protein.

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