Department of Chemistry, Purdue University, West Lafayette, IN 47907, USA.
Proteomics. 2012 Oct;12(19-20):2918-26. doi: 10.1002/pmic.201200213. Epub 2012 Sep 24.
This review presents the need for replacing gels in 2D separations for proteomics, where speed, high-throughput, and the ability to characterize trace level proteins or small samples are the current desires. The theme of the review is isoelectric focusing, which is a valuable tool because it pre-concentrates proteins in addition to separating with high peak capacity. The review traces the technological progress from gel IEF to CIEF to packed capillaries with immobilized gradients for CIEF. Multiple capillary techniques are progressing toward meeting the current desires, providing extremely high sensitivity with regard to concentration and to small samples, integrated automation, and high peak capacity from multiple dimensions of separation. Capillaries with immobilized pH gradients for CIEF are emerging, which will alleviate interference from ampholytes and improve reproducibility in separation times when this valuable technique can be used as one of the dimensions.
这篇综述提出了在蛋白质组学中取代 2D 分离中的凝胶的需求，目前的需求是速度、高通量和能够表征痕量水平的蛋白质或小样本的能力。综述的主题是等电聚焦，这是一种有价值的工具，因为它除了具有高峰容量的分离外，还可以预浓缩蛋白质。综述追溯了从凝胶 IEF 到 CIEF 再到带有固定梯度的填充毛细管的技术进步。多种毛细管技术正在朝着满足当前需求的方向发展，提供了极高的灵敏度，无论是在浓缩方面还是在小样本方面，集成自动化以及来自多个分离维度的高峰容量。用于 CIEF 的固定 pH 梯度的毛细管正在出现，这将减轻两性电解质的干扰，并提高在可以将该有价值的技术用作一个维度时的分离时间的重现性。
