杆状病毒感染的昆虫细胞中硒代蛋氨酸衍生化蛋白的生产。
Production of selenomethionyl-derivatized proteins in baculovirus-infected insect cells.
作者信息
Cronin Ciarán N, Lim Kheng B, Rogers Joe
机构信息
Pfizer, Inc., 10777 Science Center Drive, San Diego, CA 92121, USA.
出版信息
Protein Sci. 2007 Sep;16(9):2023-9. doi: 10.1110/ps.072931407. Epub 2007 Jul 27.
Abstract
A protocol is described for the production of both intracellularly expressed and secreted selenomethionyl-derivatized recombinant proteins in baculovirus-infected insect cells. The method results in the production of recombinant soluble proteins with an SeMet occupancy of approximately 75% and with a recovery of approximately 20% that of native protein expression. The method is independent of the percentage methionine content of the protein and is reliable and consistent. Similar results are obtained using either Spodoptera frugiperda Sf9 or Trichoplusia ni High Five insect cells as the expression host, and when cultures are grown in either shake flasks or in Wave BioReactors.
摘要
本文描述了一种在杆状病毒感染的昆虫细胞中生产细胞内表达和分泌的硒代蛋氨酸衍生重组蛋白的方案。该方法可产生硒代蛋氨酸占有率约为75%的重组可溶性蛋白，回收率约为天然蛋白表达的20%。该方法与蛋白质的蛋氨酸含量百分比无关，可靠且一致。使用草地贪夜蛾Sf9细胞或粉纹夜蛾High Five昆虫细胞作为表达宿主，以及在摇瓶或Wave生物反应器中培养时，均可获得类似结果。
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