Plasma lipoprotein-X quantification on filipin-stained gels: monitoring recombinant LCAT treatment ex vivo.

Journal of Lipid Research
Lita A FreemanAlan T Remaley

Abstract

Familial LCAT deficiency (FLD) patients accumulate lipoprotein-X (LP-X), an abnormal nephrotoxic lipoprotein enriched in free cholesterol (FC). The low neutral lipid content of LP-X limits the ability to detect it after separation by lipoprotein electrophoresis and staining with Sudan Black or other neutral lipid stains. A sensitive and accurate method for quantitating LP-X would be useful to examine the relationship between plasma LP-X and renal disease progression in FLD patients and could also serve as a biomarker for monitoring recombinant human LCAT (rhLCAT) therapy. Plasma lipoproteins were separated by agarose gel electrophoresis and cathodal migrating bands corresponding to LP-X were quantified after staining with filipin, which fluoresces with FC, but not with neutral lipids. rhLCAT was incubated with FLD plasma and lipoproteins and LP-X changes were analyzed by agarose gel electrophoresis. Filipin detects synthetic LP-X quantitatively (linearity 20-200 mg/dl FC; coefficient of variation <20%) and sensitively (lower limit of quantitation <1 mg/ml FC), enabling LP-X detection in FLD, cholestatic, and even fish-eye disease patients. rhLCAT incubation with FLD plasma ex vivo reduced LP-X dose dependently, generated HDL, a...Continue Reading

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Citations

Nov 7, 2019·Journal of Clinical Medicine·Itziar Lamiquiz-MoneoAna Cenarro
Feb 20, 2020·Current Opinion in Lipidology·Lita A FreemanAlan T Remaley
Oct 22, 2019·Clinical Medicine Insights. Endocrinology and Diabetes·Laura KattahCarlos O Mendivil
Jan 2, 2020·Pharmacology Research & Perspectives·Marcelo J A AmarAlan T Remaley

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