Systems for factor VIII processing and methods thereof

摘要

The present invention provides methods of reducing nonprocessed Factor VIII or a chimeric polypeptide comprising Factor VIII comprising co-transfecting in a host cell a polynucleotide encoding Factor VIII with a polynucleotide encoding a protein convertase, where the endogenous processing enzymes of the host cell are insufficient to convert all of the Factor VIII to its processed isoform; expressing a proprotein convertase from a second polynucleotide in the host cell; and reducing the nonprocessed Factor VIII by processing with said proprotein convertase.

主权项

1. A method for decreasing nonprocessed Factor VIII (FVIII) in a culturing medium, comprising contacting a nonprocessed FVIII polypeptide expressed from a polynucleotide in a host cell with proprotein convertase subtilisin/kexin type 5 (PC5),
wherein the PC5 is expressed from a second polynucleotide in the host cell expressing the FVIII, wherein the host cell is a mammalian cell, and wherein the PC5 processes the nonprocessed FVIII polypeptide to a processed FVIII polypeptide, which comprises a FVIII heavy chain and a FVIII light chain associated by a non-covalent bond.