Wilde Monika, Klausberger Miriam, Palmberger Dieter, Ernst Wolfgang, Grabherr Reingard
Vienna Institute of BioTechnology (VIBT), University of Natural Resources and Life Sciences, Muthgasse 11, 1190, Vienna, Austria,
Biotechnol Lett. 2014 Apr;36(4):743-9. doi: 10.1007/s10529-013-1429-6. Epub 2013 Dec 29.
A comparative analysis of new and established insect cell lines, in regard to process relevant parameters, provide data that can be exploited for designing more robust and effective protein production processes. The baculovirus-insect cell expression system has been efficiently used for the production of heterologous proteins. Three different insect cell lines Tnao38, High Five and Sf9 were compared in terms of virus susceptibility, baculovirus production and product yield of an intra-cellularly (YFP) and extra-cellularly (influenza A virus hemagglutinin)-expressed recombinant protein. The Tnao38 and High Five cell lines exhibited higher (tenfold) susceptibility to baculovirus infection than Sf9 cells, whereas Sf9 cells showed a higher (100-fold) capacity for production of infectious virus particles. Analysis of recombinant protein expression revealed considerably higher product yields in Tnao38 and High Five cells as compared to Sf9 cells, for both model proteins. Overall, the two Trichoplusia ni-derived cell lines, High Five and Tnao38, were significantly more efficient in terms of secreting proteins such as the glycoprotein hemagglutinin of influenza A virus.
对新的和已建立的昆虫细胞系在与工艺相关参数方面进行比较分析，可提供有助于设计更稳健、更有效的蛋白质生产工艺的数据。杆状病毒-昆虫细胞表达系统已被有效地用于生产异源蛋白。对三种不同的昆虫细胞系Tnao38、High Five和Sf9在病毒敏感性、杆状病毒产量以及细胞内（黄色荧光蛋白）和细胞外（甲型流感病毒血凝素）表达的重组蛋白的产物产量方面进行了比较。Tnao38和High Five细胞系对杆状病毒感染的敏感性比Sf9细胞高（10倍），而Sf9细胞产生传染性病毒颗粒的能力更高（100倍）。重组蛋白表达分析显示，对于两种模型蛋白，Tnao38和High Five细胞中的产物产量比Sf9细胞高得多。总体而言，两种来源于粉纹夜蛾的细胞系High Five和Tnao38在分泌诸如甲型流感病毒糖蛋白血凝素等蛋白质方面效率显著更高。
