Method of manufacturing VEGF antagonist fusion proteins
Formulations of a vascular endothelial growth factor (VEGF)-specific fusion protein antagonist are provided including a pre-lyophilized formulation, a reconstituted lyophilized formulation, and a stable liquid formulation. Preferably, the fusion protein has the sequence of SEQ ID NO:4.
1. A method of manufacturing a VEGF antagonist fusion protein, said method comprising:
a. expressing said VEGF antagonist fusion protein in a Chinese hamster ovary (CHO) cell comprising a polynucleotide encoding the VEGF antagonist fusion protein which comprises amino acids 27-457 of SEQ ID NO: 2 wherein said fusion protein binds vascular endothelial growth factor (VEGF); and
b. purifying said VEGF antagonist fusion proteins;
wherein at least 90% of the weight of the purified fusion protein is not present as an aggregate.
2. The method of claim 1 , wherein said fusion protein is purified until substantially free of protein contaminants.
3. A method of manufacturing a VEGF antagonist fusion protein, said method comprising:
a. expressing said VEGF antagonist fusion protein in a Chinese hamster ovary (CHO) cell comprising a polynucleotide encoding the VEGF antagonist fusion protein which comprises amino acids 27-457 of SEQ ID NO: 4 wherein said fusion protein binds vascular endothelial growth factor (VEGF); and
b. purifying said VEGF antagonist fusion protein;
wherein at least 90% of the weight of the purified fusion protein is not present as an aggregate.
4. The method of claim 1 , wherein said VEGF antagonist fusion protein is post transitionally glycosylated at one or more asparagine residues.