Plasma lipoprotein-X quantification on filipin-stained gels: monitoring recombinant LCAT treatment ex vivo
| Autor: | Lita A. Freeman, Sotirios K. Karathanasis, Masaki Sato, Maureen Sampson, Edward B. Neufeld, Alan T. Remaley, Robert D. Shamburek |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
lecithin:cholesterol acyltransferase Lipoproteins QD415-436 030204 cardiovascular system & hematology Biochemistry Filipin Phosphatidylcholine-Sterol O-Acyltransferase 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Endocrinology High-density lipoprotein familial lecithin:cholesterol acyltransferase deficiency Lecithin Cholesterol Acyltransferase Deficiency Methods medicine Humans Fish-Eye Disease fish-eye disease phospholipids Cholesterol cholesterol Cell Biology medicine.disease Lipoprotein-X Molecular biology Recombinant Proteins Staining 030104 developmental biology chemistry high density lipoprotein Agarose gel electrophoresis lipids (amino acids peptides and proteins) Gels Biomarkers Ex vivo Lipoprotein |
| Zdroj: | J Lipid Res Journal of Lipid Research, Vol 60, Iss 5, Pp 1050-1057 (2019) |
| ISSN: | 0022-2275 |
| Popis: | 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 |
| Databáze: | OpenAIRE |
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