Application of flow field-flow fractionation for the characterization of macromolecules of biological interest: a review

Abstract

An overview is given of the recent literature on (bio) analytical applications of flow field-flow fractionation (FlFFF). FlFFF is a liquid-phase separation technique that can separate macromolecules and particles according to size. The technique is increasingly used on a routine basis in a variety of application fields. In food analysis, FlFFF is applied to determine the molecular size distribution of starches and modified celluloses, or to study protein aggregation during food processing. In industrial analysis, it is applied for the characterization of polysaccharides that are used as thickeners and dispersing agents. In pharmaceutical and biomedical laboratories, FlFFF is used to monitor the refolding of recombinant proteins, to detect aggregates of antibodies, or to determine the size distribution of drug carrier particles. In environmental studies, FlFFF is used to characterize natural colloids in water streams, and especially to study trace metal distributions over colloidal particles. In this review, first a short discussion of the state of the art in instrumentation is given. Developments in the coupling of FlFFF to various detection modes are then highlighted. Finally, application studies are discussed and ordered according to the type of (bio) macromolecules or bioparticles that are fractionated.

Size Separation Techniques for the Characterisation of Cross-Linked Casein: A Review of Methods and Their Applications

“…The applied fields can include cross-flow stream, temperature gradient, centrifugal force, electrical potential, or gravitational force, resulting in different FFF sub-techniques that allow the fractionation of molecules according to molecular size, charge, density, and/or chemical structure. In the recent years, the theoretical background of FFF and its main applications in analysis of synthetic and natural polymers [157][158][159][160], colloids and nanoparticles [161][162][163], cells and biomolecules [159,164], and food macromolecules [165] have been reviewed in great detail.…”

Proteins and antibodies in serum, plasma, and whole blood—size characterization using asymmetrical flow field-flow fractionation (AF4)

“…Due to the absence of a stationary phase, several problems related to SEC are alleviated including minimization of non-specific protein adsorption, structural deformation at the surface and high shear forces which may result in degradation of analytes. Therefore, AF4 is a highly powerful technique that is increasingly being used for the separation and characterization of biomacromolecules and pharmaceutical molecules [14–16]. It has been proven to be a potential tool for studying biological structures such as proteins, antigens, and antibodies [17–21].…”

Proteomics - The Future of Periodontal Diagnostics

“…In addition, certain basic proteins, and very high-or very low-molecular weight proteins are not separated well by 2D-GE [7]. Another technique is Field Flow Fractionation (FFF) which separates proteins based on their mobility in presence of an applied field, such as electrical, gravitational, centrifugal etc [8].…”