We have developed a novel and more convenient LCAT activity assay using fluorescence-labelled cholesterol (Bodipy-cholesterol), which is incorporated into proteoliposomes as a substrate instead of radiolabelled cholesterol.

Recombinant human LCAT (rhLCAT) is now being developed as an enzyme replacement therapy for familial LCAT deficiency and as a possible treatment for acute coronary syndrome. The current 'gold standard' assay for LCAT activity involves the use of radioisotopes, thus making it difficult for routine clinical use.

Methods: We have developed a novel and more convenient LCAT activity assay using fluorescence-labelled cholesterol (BODIPY-cholesterol), which is incorporated into proteoliposomes as a substrate instead of radiolabelled cholesterol.

5.1. Lecithin:cholesterol acyltransferase (LCAT) assay. LCAT mediates the esterification of HDL cholesterol and favors the maintenance of a gradient of free cholesterol between the plasma membrane and HDL particles.

Labeling plasma with BODIPY-cholesterol in the presence of trehalose was found to be preferable to cryoprotect LpX and is more amenable than previous staining methods for translation into a routine diagnostic assay.

Jul 12, 2019 · We describe simple, sensitive and robust methods to monitor lipoprotein remodeling and cholesterol and apolipoprotein exchange, using fluorescent Lissamine Rhodamine B head-group tagged phosphatidylethanolamine (*PE) as a …

(B) rhLCAT or rhLCAT preincubated with NEM was incubated with rHDL containing BODIPY-cholesterol and POPC for 10-90 min at 37 °C. The extracted lipids were dissolved in 30 μL of chloroform and...

Jul 11, 2013 · Specific fluorescence detection of DHE ester synthesis is achieved by employing cholesterol oxidase to selectively render unesterified DHE non-fluorescent. The assay accurately detects LCAT...

Nov 2, 2017 · LCAT activity was a linear function of the serum sample volume up to 670 nmol/hr per ml and coefficients of variation (CV) less than 4% were obtained under the standardized conditions.

Nov 2, 2017 · We have developed a novel and more convenient LCAT activity assay using fluorescence-labelled cholesterol (BODIPY-cholesterol), which is incorporated into proteoliposomes as a substrate instead of radiolabelled cholesterol.