Patent ID: 8709756

Claim:
A method for the synthesis of a desired secreted heteromultimeric protein comprising at least two non-identical subunit polypeptide chains in a diploid Pichia yeast cell, the method comprising: (i) producing a recombinant diploid Pichia yeast by spheroplast fusion of 2 haploid Pichia spheroplasts or by mating 2 intact haploid Pichia yeast cells, resulting in a diploid Pichia yeast comprising nucleic acids encoding all of the subunits of a desired heterologous (non-yeast)-heteromultimeric, protein, which nucleic acids are each operably linked to a promoter and signal peptide that provides for the expression and secretion of the subunits encoded by the nucleic acids by said recombinant diploid Pichia yeast; and (ii) culturing said recombinant diploid Pichia yeast under conditions that provide for the expression and secretion of the desired heteromultimeric protein into a culture medium comprising the recombinant diploid Pichia yeast.