Abstract
The N-glycan patterns of recombinant human coagulation factors II (rF-II) and IX (rF-IX), derived from both transfected Chinese hamster ovary (CHO) cells and African green monkay (Vero) cells produced at industrial scale, were analyzed by binding to carbohydrate-specific lectins and were compared with the glycan structure of human plasma-derived coagulation factors. Human plasma-derived coagulation factors II (hpF-II) and IX (hpF-IX) exhibited complex-type glycan structures with carbohydrate chains capped with α(2–6)-sialic acid. Terminal galactose-β(1–4)-N-acetylglucosamine units were detected in hpF-IX. Both CHO cell-derived rF-II and rF-IX exhibited complex-type glycosylation and contained α(2–3)-sialic acid in addition to terminal galactose-β(1–4)-N-acetylglucosamine. Vero cell-derived rF-IX exhibited a complex-type glycan structure similar to that of CHO cell-derived rF-IX. In contrast, rF-II produced by Vero cells exhibited a glycan microheterogeneity composed of hybrid-type glycosylation containing “high-mannose” structures and complex-type glycosylation containing α(2–3)-sialic acid. Galactose-β(1–4)-N-acetylglucosamine structures and a low concentration of α(2–6)-sialic acid were detected in both microheterogeneity fractions of Vero cell-derived rF-II. Although different in their carbohydrate structures, coagulation factors II and IX obtained recombinantly from both transformed CHO cells and Vero cells exhibited coagulation activities comparable with the plasma-derived proteins.
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References
Kurachi K. Recombinant antihemophilic factors. In: Goldstein J, ed. Biotechnology of Blood. Boston: Butterworth-Heinemann, 1992:177–195.
Goodey AR. The production of heterologous plasma proteins. Trends Biotechnol 1993;11:430–433.
Furie B, Furie BC. The molecular basis of blood coagulation. Cell 1993;53:505–518.
Cumming DA. Physiological relevance of protein glycosylation. Dev Biol Stand 1992;76:83–94.
Geisow MJ. Glycoprotein glycans—roles and control. Trends Biotechnol 1992;10:333–335.
Rademacher TW. Glycosylation as a factor affecting product consistency. Biologicals 1993;21:103–104.
Werner RG, Noe W. Mammalian cell cultures. Genetic engineering, protein glycosylation, fermentation and process control. Drug Res 1993;43:1242–1249.
Yan SCB, Grinnell B, Wold F. Post-translational modifications of proteins. Trends Biol Sci 1989;14:264–268.
Fischer B, Mitterer A, Dorner F. Purification of recombinant human coagulation factors II and IX and protein S expressed in recombinant Vaccinia virus-infected Vero cells. J Biotechnol 1995;38:129–136.
Bradford M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976;72:248–254.
Laemmli UK. Cleavage o structural proteins during the assembly of the head of bacteriophage T4. Nature 1970;227:680–685.
Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets. Proc Natl Acad Sci USA 1979;76:4350–4354.
Paulson JC. Glycoproteins: What are the sugar chains for? Trends Biol Sci 1989;14:272–276.
Warren CE. Glycosylation. Curr Opin Biotechnol 1993;4:596–602.
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Fischer, B., Mitterer, A., Dorner, F. et al. Comparison of N-glycan pattern of recombinant human coagulation factors II and IX expressed in Chinese hamster ovary (CHO) and African green monkey (Vero) cells. J Thromb Thrombol 3, 57–62 (1996). https://doi.org/10.1007/BF00226412
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DOI: https://doi.org/10.1007/BF00226412