Review: The Glycosylation Heterogeneity of Recombinant Human IFN-γ

Abstract

The cloning of the cDNA for human interferon-gamma (IFN-gamma) has resulted in its expression in Escherichia coli, baculovirus-infected insect cells, Chinese hamster ovary (CHO) cells, and the mammary gland of transgenic mice. Large quantities of highly purified recombinant IFN-gamma have been generated, aided by the use of highly specific neutralizing monoclonal antibodies, with a view to its production as a human therapeutic protein. The primary source of structural heterogeneity for IFN-gamma during its production in mammalian expression systems is glycosylation, which can profoundly affect the three-dimensional structure of a glycoprotein and its biological function. A number of analytical approaches have been developed recently to allow a detailed analysis of the carbohydrate structures associated with IFN-gamma, the principal advances being in the areas of capillary electrophoresis and mass spectrometry. The implementation of these high-resolution analytical tools to determine the glycosylation profile of IFN-gamma makes it one of the best characterized recombinant glycoproteins. Recombinant human IFN-gamma acts as a model secretory glycoprotein, typifying the intrinsic glycosylation processing events associated with production of a potential therapeutic glycoprotein.

Analysis of carbohydrates and glycoconjugates by matrix‐assisted laser desorption/ionization mass spectrometry: An update covering the period 1999–2000

“…Methods, including MALDI, for analyzing these glycosylation differences have recently been reviewed (Hooker & James, 1998). In experiments aimed at remodeling the carbohydrates attached to human interferon-g, Fukuta et al (2000) have introduced the genes for GlcNAc transferases-IVand -V into Chinese hamster ovary (CHO) cells producing the glycoprotein.…”

The K1K2 Region of Lys-Gingipain of Porphyromonas gingivalis Blocks Induction of HLA Expression by Gamma Interferon

“…Native IFN-␥ has three forms that vary in glycosylation: diglycosylated at Asn25 and Asn97 (2N), monoglycosylated at Asn25 (1N), and nonglycosylated (12). The results suggest that the glycan side chains do not prevent the binding of K1K2 to IFN-␥.…”

Chronotherapeutics: A Novel Approach in the Pharmacotherapy of Various Diseases

“…Moreover, even the basic parameters like pH, temperature or tools used for storage and packaging can influence the final structure of the end-product [6]. For example, the glycosylation pattern of granulocytecolony stimulating factor (G-CSF) [7] and interferon-γ [8] can be different in different expression systems. Another example is EPO (erythropoietin), a molecule which gave immunogenicity issues, in some cases, due to the minor changes in the manufacturing process of the final product, although this safety issue concerns an originator [9][10][11][12]14].…”