Fluorescent assay and method that corrects for spectral interfer

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or...

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435 792, 435 79, 435 11, 435810, 435975, 436829, 436809, 436817, 514 2, C12Q 100, C12Q 260, G01N 3353

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active

055852351

ABSTRACT:
A method is provided for measuring the activity of cholesteryl ester transfer protein or MTP. The method comprises the steps of: adding a prepared emulsion particle to a buffer to form a buffered solution simulating physiological conditions, adding an emulsion of lipid to the buffered solution of prepared sonicated particle, adding a source of CETP or MTP to the buffered solution, adding a compound to the buffered solution for the purpose of testing the compound's effect on the neutral lipid transfer protein (CETP or MTP) activity, incubating the buffered mixture, reading the fluorescence of the solution, and calculating the effect of the compound on the emission spectra of the transfer label so transfer activity can than be accurately determined. A device that determines the activity of CETP or MTP by the use of a newly synthesized donor particle without regard to the presence of colored or otherwise interfering factors. A system comprises a donor particle comprised of a self quenching fluorescent neutral lipid core, an acceptor particle to accept protein facilitated transfer of fluorescent neutral lipid, and determining interference on the emission intensity of the fluorescence of the particles.

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