2.205 School of Biological Sciences, University of Manchester, Stopford Building, Oxford Road, Manchester, M13 9PT (Author for correspondence).
Cytotechnology. 2000 Feb;32(2):109-23. doi: 10.1023/A:1008170710003.
The production of recombinant proteins using mammalian cell expression systems is of growing importance within biotechnology, largely due to the ability of specific mammalian cells to carry out post-translational modifications of the correct fidelity. The Glutamine Synthetase-NS0 system is now one such industrially important expression system.Glutamine synthetase catalyses the formation ofglutamine from glutamate and ammonia. NS0 cellscontain extremely low levels of endogenous glutaminesynthetase activity, therefore exogenous glutaminesynthetase can be used efficiently as a selectablemarker to identify successful transfectants in theabsence of glutamine in the media. In addition, theinclusion of methionine sulphoximine, an inhibitor ofglutamine synthetase activity, enables furtherselection of those clones producing relatively highlevels of transfected glutamine synthetase and henceany heterologous gene which is coupled to it. Theglutamine synthetase system technology has been usedfor research and development purposes during thisdecade and its importance is clearly demonstrated nowthat two therapeutic products produced using thissystem have reached the market place.
使用哺乳动物细胞表达系统生产重组蛋白在生物技术中变得越来越重要,主要是因为特定的哺乳动物细胞能够以正确的保真度进行翻译后修饰。谷氨酰胺合成酶 - NS0 系统现在是这样一种工业上重要的表达系统。谷氨酰胺合成酶催化谷氨酸和氨形成谷氨酰胺。NS0 细胞内源性谷氨酰胺合成酶活性极低,因此外源性谷氨酰胺合成酶可以有效地用作选择标记物,在培养基中没有谷氨酰胺的情况下鉴定成功转染的细胞。此外,加入甲硫氨酸亚砜imine,一种谷氨酰胺合成酶活性的抑制剂,可以进一步选择那些产生相对高水平转染谷氨酰胺合成酶的克隆,以及与之偶联的任何异源基因。谷氨酰胺合成酶系统技术在过去十年中一直用于研究和开发目的,现在已经有两种使用该系统生产的治疗产品进入市场,其重要性显而易见。