Hooker A, James D
Oxford GlycoSciences (UK) Plc, Abingdon Science Park, United Kingdom.
J Interferon Cytokine Res. 1998 May;18(5):287-95. doi: 10.1089/jir.1998.18.287.
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.
人γ干扰素（IFN-γ）cDNA的克隆使其能在大肠杆菌、杆状病毒感染的昆虫细胞、中国仓鼠卵巢（CHO）细胞以及转基因小鼠的乳腺中表达。借助高度特异性的中和单克隆抗体，已大量生产出高度纯化的重组IFN-γ，目的是将其作为一种人类治疗性蛋白质来生产。在哺乳动物表达系统中生产IFN-γ时，其结构异质性的主要来源是糖基化，糖基化会深刻影响糖蛋白的三维结构及其生物学功能。最近已开发出多种分析方法，以便对与IFN-γ相关的碳水化合物结构进行详细分析，主要进展集中在毛细管电泳和质谱领域。运用这些高分辨率分析工具来确定IFN-γ的糖基化谱，使其成为特征最明确的重组糖蛋白之一。重组人IFN-γ作为一种典型的分泌型糖蛋白，代表了与潜在治疗性糖蛋白生产相关的内在糖基化加工过程。
