Abstract
As an increasing number of recombinant therapeutic glycoproteins are manufactured and investigated, the importance of their attached glycans is becoming more widely reported and understood. Regulatory agencies expect detailed “extended characterization” of the glycoprotein as well as routine, well-controlled “release assays” with specifications to be employed for quality control of each manufactured lot. In this chapter we will briefly discuss relevant glycan issues in the area of therapeutic recombinant glycoprotein manufacture and describe in detail two assays that are employed in the development of, for example, recombinant Factor VIII for the treatment of hemophilia.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Hossler P, Khattak SF, Li ZJ (2009) Optimal and consistent protein glycosylation in mammalian cell culture. Glycobiology 19:936–949
Raju TS (2003) Glycosylation variations with expression systems and their impact on biological activity of therapeutic immunoglobulins. Bioprocess Int 1:44–53
Davies J, Jiang L, Pan LZ, LaBarre MJ, Anderson D, Reff M (2001) Expression of GnTIII in a recombinant anti-CD20 CHO production cell line: expression of antibodies with altered glycoforms leads to an increase in ADCC through higher affinity for FC gamma RIII. Biotechnol Bioeng 74:288–294
Bork K, Horstkorte R, Weidemann W (2009) Increasing the sialylation of therapeutic glycoproteins: the potential of the sialic acid biosynthetic pathway J. Pharm Sci.98:3499–3508
Bovenschen N, Rijken DC, Havekes LM, Van Vlijmen BJ, Mertens M (2005) The B domain of coagulation factor VIII interacts with the asialoglycoprotein receptor. J Thromb Haemost 3:1257–1265
Solá R, Griebenow K (2010) Glycosylation of therapeutic proteins. An effective strategy to optimize efficacy. BioDrugs 24:9–21
Purcell RT, Lockey RF (2008) Immunologic responses to therapeutic biologic agents. J Investig Allergol Clin Immunol 18:335–343
Chung CH, Mirakhur B, Chan E, Le QT, Berlin J, Morse M, Murphy BA, Satinover SM, Hosen J, Mauro D, Slebos RJ, Zhou Q, Gold D, Hatley T, Hicklin DJ, Platts-Mills TA (2008) Cetuximab-induced anaphylaxis and IgE specific for galactose-alpha-1,3-galactose. N Engl J Med 358:1109–1117
Higgins E (2010) Carbohydrate analysis throughout the development of a protein therapeutic. Glycoconj J 27:211–225
Fang H, Wang L, Wang H (2007) The protein structure and effect of factor VIII. Thromb Res 119:1–13
White G, Pickens E, Liles D, Roberts H (1998) Mammalian recombinant coagulation proteins: structure and function. Transfus Sci 19:177–189
Anumula KR, Dhume ST (1998) High resolution and high sensitivity methods for oligosaccharide mapping and characterization by normal phase high performance liquid chromatography following derivatization with highly fluorescent anthranilic acid. Glycobiology 8:685–694
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Samuels, N., Kates, D., Liu, J., Severs, J. (2013). Characterizing the Glycosylation State of Therapeutic Recombinant Glycoproteins. In: Kohler, J., Patrie, S. (eds) Mass Spectrometry of Glycoproteins. Methods in Molecular Biology, vol 951. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-146-2_22
Download citation
DOI: https://doi.org/10.1007/978-1-62703-146-2_22
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-145-5
Online ISBN: 978-1-62703-146-2
eBook Packages: Springer Protocols