Microtech2010 2010

Hollow-fiber flow field-flow fractionation: a novel pre-MS method for proteomics

D. Rambaldi, A. Zattoni, P. Reschiglian, C. Johann
University of Bologna, IT

Keywords: hollow-fiber flow field-flow fractionation, mass spectrometry, Pre-MS separation methods, proteomics


It is well acknowledged that both number and dynamic range of the proteins present in biological systems lead to the need of a “simplified” and more efficient way to proteome analysis. The availability of so-called “pre-MS” methods for protein isolation/separation from complicated biological samples is thus strictly required for successful MS-based approaches to current proteomics. Hollow-fiber flow field-flow fractionation (HF5) is the innovative, ready-to-market microcolumn version of flow field-flow fractionation (F4). HF5 is ideally suited as online or offline pre-MS separation step within a comprehensive analytical platform for the characterization of complex protein samples in native form. We have developed different applications based on HF5 in combination with MS or LC-MS for the fractionation/purification and characterization of native proteins, and for proteomic analysis of complex protein samples. When applied to the analysis of subproteomes associated with nanosized biological structures such as lipoproteins (LPs) in blood serum, HF5 shows promising for the development of methods based on integrated techniques to identify not only apo-lipoproteins in the different LP classes, but also LP/low-abundance protein (LAP) complexes.
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