Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal.
Talanta. 2010 Feb 15;80(4):1561-8. doi: 10.1016/j.talanta.2009.06.081. Epub 2009 Jul 9.
A new application of intact cell MALDI-TOF MS (ICM-MS) methodology is described for monitoring the production of viral proteins and viral like particles using the baculovirus/insect cells expression system. Various MALDI matrices, cell preparation methods, cell/matrix volume ratio and MALDI target application procedures were tested in order to obtain the highest intensity and reproducibility of intact insect cell spectra. The web interface, SPECLUST (http://bioinfo.thep.lu.se/speclust.html), was used to construct dendograms based on MALDI-TOF MS data for evaluation of fingerprint changes. We demonstrate that insect cell mass spectrum fingerprints are characteristic of each viral protein/particle production. Their changes along the time for each production experiment correlate with the intracellular viral protein content determined by Western blot. This work shows that this simple, fast and low cost assay, which requires low sample volume, is a powerful analytical tool that complements the most common analytical methods used for monitoring bioprocesses and has potential application in the biotechnological industry namely, in the production of recombinant proteins.
一种新的应用完整细胞 MALDI-TOF MS（ICM-MS）方法被描述用于监测使用杆状病毒/昆虫细胞表达系统生产病毒蛋白和类似病毒颗粒。为了获得最强的完整昆虫细胞光谱强度和重现性，测试了各种 MALDI 基质、细胞制备方法、细胞/基质体积比和 MALDI 靶应用程序。使用网络界面 SPECLUST（http://bioinfo.thep.lu.se/speclust.html）根据 MALDI-TOF MS 数据构建 dendograms，用于评估指纹变化。我们证明，昆虫细胞质谱指纹图谱是每种病毒蛋白/颗粒生产的特征。它们在每个生产实验中的变化与通过 Western blot 确定的细胞内病毒蛋白含量相关。这项工作表明，这种简单、快速且低成本的测定方法需要少量样品，是一种强大的分析工具，可补充用于监测生物工艺的最常见分析方法，并具有在生物技术行业中的潜在应用，即重组蛋白的生产。
