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Rapid two-dimensional characterisation of proteins in solution
Microsystems & Nanoengineering ( IF 7.3 ) Pub Date : 2019-07-01 , DOI: 10.1038/s41378-019-0072-3
Kadi L Saar 1 , Quentin Peter 1, 2 , Thomas Müller 3 , Pavan K Challa 1, 2 , Therese W Herling 1 , Tuomas P J Knowles 1, 2
Affiliation  

Microfluidic platforms provide an excellent basis for working with heterogeneous samples and separating biomolecular components at high throughput, with high recovery rates and by using only very small sample volumes. To date, several micron scale platforms with preparative capabilities have been demonstrated. Here we describe and demonstrate a microfluidic device that brings preparative and analytical operations together onto a single chip and thereby allows the acquisition of multidimensional information. We achieve this objective by using a free-flow electrophoretic separation approach that directs fractions of sample into an on-chip analysis unit, where the fractions are characterised through a microfluidic diffusional sizing process. This combined approach therefore allows simultaneously quantifying the sizes and the charges of components in heterogenous mixtures. We illustrate the power of the platform by describing the size distribution of a mixture comprising components which are close in size and cannot be identified as individual components using state-of-the-art solution sizing techniques on their own. Furthermore, we show that the platform can be used for two-dimensional fingerprinting of heterogeneous protein mixtures within tens of seconds, opening up a possibility to obtain multiparameter data on biomolecular systems on a minute timescale.



中文翻译:

溶液中蛋白质的快速二维表征

微流体平台为处理异质样品和以高通量、高回收率和仅使用非常小的样品体积分离生物分子成分提供了极好的基础。迄今为止,已经展示了几个具有制备能力的微米级平台。在这里,我们描述并展示了一种微流体装置,它将制备和分析操作结合到一个芯片上,从而允许获取多维信息。我们通过使用自由流动的电泳分离方法来实现这一目标,该方法将样品的部分引导到芯片分析单元中,其中部分通过微流体扩散大小调整过程进行表征。因此,这种组合方法允许同时量化异质混合物中组分的大小和电荷。我们通过描述混合物的尺寸分布来说明该平台的功能,该混合物包含尺寸相近的成分,并且不能使用最先进的解决方案尺寸技术单独识别为单个成分。此外,我们表明该平台可用于在数十秒内对异质蛋白质混合物进行二维指纹识别,为在分钟时间尺度上获得生物分子系统的多参数数据开辟了可能性。我们通过描述混合物的尺寸分布来说明该平台的功能,该混合物包含尺寸相近的成分,并且不能使用最先进的解决方案尺寸技术单独识别为单个成分。此外,我们表明该平台可用于在数十秒内对异质蛋白质混合物进行二维指纹识别,为在分钟时间尺度上获得生物分子系统的多参数数据开辟了可能性。我们通过描述混合物的尺寸分布来说明该平台的功能,该混合物包含尺寸相近的成分,并且不能使用最先进的解决方案尺寸技术单独识别为单个成分。此外,我们表明该平台可用于在数十秒内对异质蛋白质混合物进行二维指纹识别,为在分钟时间尺度上获得生物分子系统的多参数数据开辟了可能性。

更新日期:2019-11-18
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