PH20 polypeptide variants, formulations and uses thereof

A method for manufacture of a protein, comprising preparing a plasmid DNA containing a cDNA encoding the protein comprising an amino acid sequence, wherein:

• (a) the protein is a modified PH20 polypeptide;
• (b) at least 95% of the residues of the amino acid sequence of the encoded modified PH20 polypeptide, excluding any signal sequence, are identical to the residues in an amino acid sequence selected from the group consisting of SEQ ID NO:
• 3 and 32-66 when the sequence of the modified PH20 polypeptide is aligned at positions corresponding to the sequence selected from the group consisting of SEQ ID NO:
• 3 and 32-66 to maximize identical residues, and wherein terminal gaps are treated as non-identical;
• and
• (c) the amino acid sequence of the modified PH20 polypeptide comprises an amino acid modification at a position corresponding to position 320 with reference to amino acid positions set forth in SEQ ID NO:
• 3, and wherein the modification at position 320 is selected from among H, K, Rand S;
• wherein the method comprises:
• transfecting the plasmid into a host cell;
• culturing the cell under conditions for expression of the cDNA as a protein;
• harvesting the protein from the culture.