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在CHO细胞和昆虫细胞中生产吸血蝙蝠纤溶酶原激活剂DSPAα1。

Production of vampire bat plasminogen activator DSPA alpha 1 in CHO and insect cells.

作者信息

Petri T, Langer G, Bringmann P, Cashion L, Shallow S, Schleuning W D, Donner P

机构信息

Research Laboratories of Schering AG, Berlin, Germany.

出版信息

J Biotechnol. 1995 Feb 21;39(1):75-83. doi: 10.1016/0168-1656(94)00146-4.

Abstract

Salivary plasminogen activator from the vampire bat Desmodus rotundus (DSPA alpha 1) is a promising new thrombolytic agent. Continuous growth of a stably transfected, methotrexate amplified, dhfr- CHO cell line yields up to 60 mg l-1 of DSPA alpha 1. Utilizing an engineered baculovirus 10 mg l-1 were produced in batches of Sf9 insect cells. Recombinant DSPA alpha 1 is purified from both sources using a one-step purification protocol. Although differences in glycosylation were detected, enzymatic activity and fibrin cofactor dependency are unaffected when DSPA alpha 1 derived from the two expression systems is compared.

摘要

吸血蝙蝠(Desmodus rotundus)的唾液纤溶酶原激活剂(DSPAα1)是一种很有前景的新型溶栓剂。稳定转染、经甲氨蝶呤扩增的dhfr-CHO细胞系持续培养可产生高达60毫克/升的DSPAα1。利用一种工程杆状病毒,在Sf9昆虫细胞批次培养中可产生10毫克/升。重组DSPAα1可通过一步纯化方案从这两种来源中进行纯化。尽管检测到糖基化存在差异,但比较源自两种表达系统的DSPAα1时,其酶活性和对纤维蛋白辅因子的依赖性不受影响。

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