title: Recombinant human factor IX produced from transgenic porcine milk.
abstract: Production of biopharmaceuticals from transgenic animal milk is a cost-effective method for highly complex proteins that cannot be efficiently produced using conventional systems such as microorganisms or animal cells. Yields of recombinant human factor IX (rhFIX) produced from transgenic porcine milk under the control of the bovine α-lactalbumin promoter reached 0.25 mg/mL. The rhFIX protein was purified from transgenic porcine milk using a three-column purification scheme after a precipitation step to remove casein. The purified protein had high specific activity and a low ratio of the active form (FIXa). The purified rhFIX had 11.9 γ-carboxyglutamic acid (Gla) residues/mol protein, which approached full occupancy of the 12 potential sites in the Gla domain. The rhFIX was shown to have a higher isoelectric point and lower sialic acid content than plasma-derived FIX (pdFIX). The rhFIX had the same N-glycosylation sites and phosphorylation sites as pdFIX, but had a higher specific activity. These results suggest that rhFIX produced from porcine milk is physiologically active and they support the use of transgenic animals as bioreactors for industrial scale production in milk.
authors: Division of Animal Technology, Animal Technology Laboratories, Agricultural Technology Research Institute, No. 52 Kedong 2 Road, Zhunan Township, Miaoli County 350, Taiwan.
authors: Meng-Hwan
authors: MH
authors: Lee
authors: Division of Animal Technology, Animal Technology Laboratories, Agricultural Technology Research Institute, No. 52 Kedong 2 Road, Zhunan Township, Miaoli County 350, Taiwan.
authors: Yin-Shen
authors: YS
authors: Lin
authors: Division of Animal Technology, Animal Technology Laboratories, Agricultural Technology Research Institute, No. 52 Kedong 2 Road, Zhunan Township, Miaoli County 350, Taiwan.
authors: Ching-Fu
authors: CF
authors: Tu
authors: Division of Animal Technology, Animal Technology Laboratories, Agricultural Technology Research Institute, No. 52 Kedong 2 Road, Zhunan Township, Miaoli County 350, Taiwan.
authors: Chon-Ho
authors: CH
authors: Yen
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D019149:Bioreactors; D002417:Cattle; D005164:Factor IX / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D008892:Milk / Q000737:chemistry* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D013552:Swine
chemical_list: D011994:Recombinant Proteins; D005164:Factor IX