Analytical Strategies for Characterisation and Quantitation of Zinc-Binding Proteins by 2D-LC coupled to ICP-MS, ESI-Q-TOF-MS and MALDI-TOF-TOF-MS

摘要

Pre-fractionation and separation is critical for successful mass spectrometric detection of biomolecules in complex sample types. In recent years there has been increasing interest in the analysis of intact proteins, and a number of novel column types have been introduced to address this. Monolithic columns offer superior mass transfer properties, as compared to traditional packed columns, and thus are well suited to separation of intact protein species. A widely used approach for increasing the selectivity of a separation system is the use of two or more separation stages in series, resulting in a multidimensional separation. The combination of ion exchange (IE) HPLC and reverse phase (RP) HPLC, employing a monolithic column, offers a high separation capacity for intact proteins in relatively complex samples. This strategy has been employed here for the study of intact proteins in bacterial cell lysates.