1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC; Avanti Polar Lipids, Alabaster, AL, USA) was formed into 100 nm multilamellar vesicles (MLV) via extrusion using the LiposoFast system (Avestin, Ottawa, ON, Canada) as previously described [7 (link)]. MLV were incubated with RG54 or ApoA-I at the indicated ratios for 4 days at 24°C to form rHDL. The resulting particle sizes were measured by dynamic light scattering (DLS) using a Zetasizer APS (Malvern Instruments, Uppsala, Sweden).
Turbid MLV solutions at 30 nmol/l DMPC were combined with PBS, RG54 or ApoA-I at a 1:100 molar ratio, and lipid binding at 25°C was measured by absorbance at 325 nm at the indicated times. Readings were fitted to one-way decay of non-linear regression.
Turbid MLV solutions at 30 nmol/l DMPC were combined with PBS, RG54 or ApoA-I at a 1:100 molar ratio, and lipid binding at 25°C was measured by absorbance at 325 nm at the indicated times. Readings were fitted to one-way decay of non-linear regression.
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