Miniaturization of asymmetrical flow field-flow fractionation and application to studies on lipoprotein aggregation and fusion

Protein aggregation: Pathways, induction factors and analysis

“…The resolution of high molecular weight (HMW) molecules is easily achieved due to the low diffusion coefficient driving them closer to the accumulation wall by the movement of the cross-flow and eluting out slower whilst opposite is true for the low molecular weight (LMW) molecules having a high diffusivity. 112,116 An elementary association between the experimental retention time and the particle sizes of samples can be predicted based on the rate of diffusion of the particles by a theory described elsewhere.…”

Conformational changes of apoB-100 in SMase-modified LDL mediate formation of large aggregates at acidic pH

“…Aggregation of the LDL samples was followed by measuring the absorbance of the LDL samples at 405 nm. The sizes of the aggregated particles were determined by asymmetrical fl ow-fi eld-fl ow fractionation (AsFlFFF) as described ( 24 ) or by dynamic light scattering (DLS) (Zetasizer Nano, Malvern). 35 S-radiolabeled LDL particles and 3 H-radiolabeled lipid microemulsions, either unmodifi ed or bcSMase-modifi ed, were incubated together in the pH buffers as described above.…”

“…The amounts of LDL were expressed in terms of their protein concentrations, which were determined by the method of Lowry et al ( 20 ) AsFlFFF, a method that has been used to determine the sizes of modifi ed LDL particles ( 24 ). We fi rst treated the lipid microemulsions with bcSMase at neutral pH, stopped the activity of the enzyme, and divided the bcSMase-treated lipid microemulsions into two aliquots, whose pH was adjusted to 5.5 or 7.5.…”

Comparison of Miniaturized and Conventional Asymmetrical Flow Field-Flow Fractionation (AF4) Channels for Nanoparticle Separations

“…Afterwards, a miniaturized AF4 channel without inlet frit was introduced by Riekkola et al in 2006. It was demonstrated that this miniaturized channel was suitable for protein standard separation and was comparable to the conventional channel [21]. In 2011, Kim et al developed a new chip-type AF4 channel design, which was assembled by stacking multilayers of thin stainless steel plates and a frit.…”