Novel liquid-chromatography columns for proteomics research

The enormous interest in proteomics research in recent years has inspired many developments in peptide chromatography. Different strategies have been developed to cope with the vast complexity of proteomics samples, trying to provide sufficient degree of separation to be able to exploit fully the potential of protein identification by mass spectrometry (MS). As reversed-phase liquid chromatography (RPLC) coupled to MS is still the method of choice for the analysis of protein digests, many efforts focus on the development of high-efficiency RP methods (e.g., monolithic columns and ultra-high-performance LC). This can also increase the speed and the sensitivity of the analysis of protein digests.As RPLC-MS alone is unlikely to provide sufficient resolution to unravel the composition of highly complex samples comprehensively, multidimensional methods will remain essential in proteome research. In this area, hydrophilic interaction chromatography (HILIC) seems to be a promising alternative to the traditional strong cation-exchange-based methods. Also, HILIC has found application in the analysis of post-translational modifications (e.g., phosphorylation and glycosylation).This review describes recent developments in LC methods for proteomics research, focusing on advances in column technology and the application of novel column materials. Illustrative examples show the possibilities of the new columns in proteomics research.

► New LC methods are based on ultra-high performance liquid chromatography. ► Monolithic & open-tubular columns can increase the efficiency of protein digest analysis. ► HILIC is very suitable for the analysis of post-translational modifications. ► Multidimensional separation systems are important for complex proteomics samples.