Flow field-flow fractionation (FlFFF) is a versatile size-based separation method suitable for biological macromolecules including proteins/protein aggregates, DNA, subcellular organelles, extracellular species, and whole cells. This review introduces briefly the basic principles of FlFFF and its recent applications for proteomic and lipidomic analysis, which are described in two parts: (1) off-line coupling of FlFFF with MS and other bioanalytical methods, and (2) on-line FlFFF with MS. The first part includes applications for lipoproteins, exosomes, and subcellular organelles for the size-dependent analysis of proteins and lipids in narrow size-fractions collected during FlFFF, followed by independent analysis including western blotting and nanoflow liquid chromatography-electrospray ionisation-tandem mass spectrometry (nLC-ESI-MS/MS). The second part highlights the on-line FlFFF-MS, in which a miniaturised FlFFF channel is coupled to ESI-MS/MS for the high-speed lipid analysis of lipoproteins and to inductively-coupled plasma MS for the direct analysis of metals in metalloproteins from blood plasma.
All Science Journal Classification (ASJC) codes
- Analytical Chemistry