Analysis of the N-glycans of recombinant human Factor IX purified from transgenic pig milk.
Glycosylation of recombinant proteins is of particular importance because it can play significant roles in the clinical properties of the glycoprotein. In this work, the N-glycan structures of recombinant human Factor IX (tg-FIX) produced in the transgenic pig mammary gland were determined. The majority of the N-glycans of transgenic pig-derived Factor IX (tg-FIX) are complex, bi-antennary with one or two terminal N-acetylneuraminic acid (Neu5Ac) moieties. We also found that the N-glycan structures of tg-FIX produced in the porcine mammary epithelial cells differed with respect to N-glycans from glycoproteins produced in other porcine tissues. tg-FIX contains no detectable Neu5Gc, the sialic acid commonly found in porcine glycoproteins produced in other tissues. Additionally, we were unable to detect glycans in tg-FIX that have a terminal Galalpha(1,3)Gal disaccharide sequence, which is strongly antigenic in humans. The N-glycan structures of tg-FIX are also compared to the published N-glycan structures of recombinant human glycoproteins produced in other transgenic animal species. While tg-FIX contains only complex structures, antithrombin III (goat), C1 inhibitor (rabbit), and lactoferrin (cow) have both high mannose and complex structures. Collectively, these data represent a beginning point for the future investigation of species-specific and tissue/cell-specific differences in N-glycan structures among animals used for transgenic animal bioreactors.
Gil
Geun-Cheol
GC
Department of Chemical and Biomolecular Engineering, University of Nebraska-Lincoln, Lincoln, NE 68588, USA.
Velander
William H
WH
Van Cott
Kevin E
KE
D000818:Animals; D030801:Animals, Genetically Modified; D000990:Antithrombin III / Q000737:chemistry; D002417:Cattle; D050718:Complement C1 Inhibitor Protein / Q000737:chemistry; D005164:Factor IX / Q000096:biosynthesis / Q000737:chemistry*; D005260:Female; D006031:Glycosylation; D006041:Goats; D006801:Humans; D007781:Lactoferrin / Q000737:chemistry; D008321:Mammary Glands, Animal / Q000378:metabolism; D008892:Milk / Q000737:chemistry*; D019158:N-Acetylneuraminic Acid / Q000737:chemistry; D011134:Polysaccharides / Q000737:chemistry*; D011247:Pregnancy; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D013045:Species Specificity; D034421:Sus scrofa
D050718:Complement C1 Inhibitor Protein; D011134:Polysaccharides; D011994:Recombinant Proteins; D000990:Antithrombin III; D005164:Factor IX; D007781:Lactoferrin; D019158:N-Acetylneuraminic Acid