Field flow fractionation in biomedical analysis

Abstract

Samples of biomedical interest which have been analysed by field-flow fractionation techniques are surveyed. The list begins with whole cells and microorganisms, going through viruses, nucleic acids, cell fragments and organelles, down to proteins and their aggregates. The principles of separation in the normal and steric mode of retention are illustrated, and instrumentation and techniques are described. The review concentrates mainly on the two systems of choice for biomedical applications: sedimentation and flow field-flow fractionation.

Sedimentation Field-Flow Fractionation of Cellular Species

“…The first is sized particles. This form of stop flow injection is de- that most of our previous experiences with RBC and bility to the external field, the lifting forces act on the GFFF (18,19,24) were gained on channels of similar particles in motion along the channel to counteract the thickness and such design may allow GFFF and SdFFF field effect and drive the particles away from the accuelution comparison. The second is a consequence of the mulation wall (27).…”

“…Every day, the chanhyperlayer (1, [18][19][20]. In such a model, assuming the nel was flushed (0.1 ml/min) for 1 hr with Coulter cleanbasic hypothesis that no particle-wall interaction may ing agent for deproteinization of the Lexan plates and occur, eluted particles are submitted to the external then washed with freshly doubly distilled water (0.5 field and to flow-rate-dependent lifting forces (1, 11).…”

“…In each of the injection device with a 20-mL loop. A Gilson Model 302 following experiments, the system void volume was dechromatographic pump (Champigneulles, France) alfined by elution of blood proteins as described in previous lowed the mobile phase to flow through the SdFFF deexperiments (18,19). Retention volumes of the eluted popvice.…”

High‐Performance Liquid Chromatography of Biological Macromolecules

“…An example is ''fast protein liquid chromatography'' (FPLC), which differs from regular HPLC mainly by being performed on instrumentation designed to suit the particular needs of biochemists. There are also techniques that are adjunct to HPLC, such as capillary electrochromatography (CEC), 16 -19 micellar electrokinetic chromatography (MEKC), 20 -23 countercurrent chromatography (CCC), 24 -26 and some modes of field flow fractionation (FFF), 27,28 which could have been part of this overview. However, this treatise will have to limit itself to hydraulically driven techniques employing a liquid eluent, and a rigid solid or gel as a stationary phase.…”

Scientific Considerations for Generic Synthetic Salmon Calcitonin Nasal Spray Products

“…This means that the sizes of peptide aggregates in the proposed generic product are comparable to those in the RLD, and that their corresponding levels are lower or the same as the levels observed in the RLD. In addition, it is important to evaluate aggregate formation by using orthogonal techniques, including (but not limited to) size exclusion chromatography, analytical ultracentrifugation (AUC), NMR, and field flow fractionation (33)(34)(35)(36), to cover all relevant aspects of aggregate size. Furthermore, evaluation of sub-visible particulates, using USP <788> and/or other methods to evaluate sub-visible particulates in the size range of 2-10 μm, can be performed to ensure comparability of particles that may affect immunogenicity.…”