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Advances in animal cell recombinant protein production: GS-NS0 expression system

机译:动物细胞重组蛋白生产的进展:GS-NS0表达系统

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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.
机译:使用哺乳动物细胞表达系统生产重组蛋白在生物技术中越来越重要,这主要是由于特定的哺乳动物细胞能够对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由谷氨酸和氨形成谷氨酰胺。 NS0细胞含有极低水平的内源性谷氨酰胺合成酶活性,因此外源谷氨酰胺合成酶可以有效地用作选择标记,以在培养基中不存在谷氨酰胺的情况下鉴定成功的转染子。另外,由于包含了谷氨酰胺合成酶活性的抑制剂蛋氨酸硫肟亚胺,因此能够进一步选择产生相对高水平转染的谷氨酰胺合成酶的克隆,并因此选择与其偶联的任何异源基因。在这个十年中,谷氨酰胺合成酶系统技术已被用于研究和开发目的,由于使用该系统生产的两种治疗产品已经进入市场,因此其重要性已得到明确证明。

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