Combariza Marianny Y, Savariar Elamprakash N, Vutukuri Dharma Rao, Thayumanavan S, Vachet Richard W
Department of Chemistry, University of Massachusetts, Amherst, Massachusetts 01003, USA.
Anal Chem. 2007 Sep 15;79(18):7124-30. doi: 10.1021/ac071001d. Epub 2007 Aug 11.
Analyses of peptides in complex mixtures are significant challenges in proteomics applications. Here, we report an amphiphilic polymer-based nanoassembly that is capable of selectively extracting peptides, on the basis of their isoelectric points, into an immiscible organic phase from an aqueous solution. The isoelectric point (pI) cutoff in these extractions depends on the pH of the aqueous solution, and thus, sequential fractionation of peptide mixtures based on pI can be accomplished by varying the pH of the aqueous solution. Additionally, we observe an unexpected enhancement in the MALDI-MS signal for extracted peptides ionized in the presence of the polymer, which allows us to obtain reproducible ion signals for some peptides at concentrations as low as 10 pM.
在蛋白质组学应用中,分析复杂混合物中的肽段是一项重大挑战。在此,我们报告了一种基于两亲聚合物的纳米组装体,它能够根据肽段的等电点,将其从水溶液中选择性地萃取到不混溶的有机相中。这些萃取过程中的等电点(pI)截止值取决于水溶液的pH值,因此,通过改变水溶液的pH值,可以实现基于pI的肽段混合物的顺序分级分离。此外,我们观察到在聚合物存在下电离的萃取肽段的基质辅助激光解吸电离质谱(MALDI-MS)信号出现意外增强,这使我们能够在低至10 pM的浓度下获得一些肽段的可重现离子信号。