High-Throughput Mass Spectrometry Applied to Structural Genomics

Georgina Berridge; Nicola A. Burgess-Brown; Rod Chalk

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High-Throughput Mass Spectrometry Applied to Structural Genomics

机译：高通量质谱技术在结构基因组学中的应用

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Mass spectrometry (MS) remains under-utilized for the analysis of expressed proteins because it is inaccessible to the non-specialist, and sample-turnaround from service labs is slow. Here, we describe 3.5 min Liquid-Chromatography (LC)-MS and 16 min LC-MSMS methods which are tailored to validation and characterization of recombinant proteins in a high throughput structural biology pipeline. We illustrate the type and scope of MS data typically obtained from a 96-well expression and purification test for both soluble and integral membrane proteins (IMPs), and describe their utility in the selection of constructs for scale-up structural work, leading to cost and efficiency savings. We propose that value of MS data lies in how quickly it becomes available and that this can fundamentally change the way in which it is used.

机译：质谱（MS）仍未充分利用来分析表达的蛋白质，因为非专业人员无法使用它，并且服务实验室的样品周转速度很慢。在这里，我们描述了3.5分钟的液相色谱（LC）-MS和16分钟的LC-MSMS方法，这些方法专门用于验证和表征高通量结构生物学管道中的重组蛋白。我们说明了通常从96孔可溶性和整合膜蛋白（IMP）的96孔表达和纯化测试中获得的MS数据的类型和范围，并描述了它们在选择用于扩大结构工作的构建体中的效用，从而导致成本和效率节省。我们认为，MS数据的价值在于它变得可用的速度，并且可以从根本上改变其使用方式。

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《Chromatography》 |2014年第4期|共17页
作者
Georgina Berridge; Nicola A. Burgess-Brown; Rod Chalk;
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中图分类生物物理学;
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High-Throughput Mass Spectrometry Applied to Structural Genomics

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