Direct isolation of labeled low density lipoproteins for the determination of cholesteryl ester transfer protein activity.
The measurement of the activity of cholesteryl ester transfer protein (CETP), is of high clinical interest and this study reports the use of a direct LDL isolation (d-LDL) technique to determine in one step the amount of radiolabeled cholesteryls esters ([3H]-CE) transferred from exogenous HDL3 to LDL, avoiding the conveniences of the usually used ultracentrifugation or precipitation of apo-B containing lipoproteins in the CETP methodologies. The d-LDL technique providing a specific immunoprecipitation of VLDL, IDL and HDL allowed to directly determine the [3H]-CE transferred on LDL (d-[3H]-CE-LDL). Two methodologies were assayed for the CETP activity using either exogenous or endogenous lipoproteins, and the results with the d-LDL technique were compared with those obtained using the ultracentrifugation (u-[3H]-CE-LDL) considered as the reference method. The intra- and inter-assays were similar in both techniques for the two CETP activity assays. Strong positive correlations were established between values obtained with d-[3H]-CE-LDL and u-[3H]-CE-LDL isolation procedures for CETP activities with exogenous or endogenous lipoproteins (r = 0.972; p = 0.0001 and r = 0.965; p = 0.0001 respectively). In conclusion, the d-LDL technique represents an easy and accurate procedure to measure directly, in normotriglyceridemic plasmas, the amount of [3H]-CE transferred from HDL to LDL by the CETP.
Sich
D
D
Laboratoire de Biochemie des Lipides, Hôpital de la Pitìé, Paris, France.
Saïdi
Y
Y
Egloff
M
M
Giral
P
P
Gautier
V
V
Federspiel
M C
MC
Turpin
G
G
Beucler
I
I
D002352:Carrier Proteins / Q000097:blood*; D053480:Cholesterol Ester Transfer Proteins; D002788:Cholesterol Esters / Q000097:blood; D008078:Cholesterol, LDL / Q000097:blood; D006023:Glycoproteins; D006801:Humans; D006937:Hypercholesterolemia / Q000097:blood; D008077:Lipoproteins, LDL / Q000097:blood* / Q000302:isolation & purification*; D012016:Reference Values; D014316:Tritium; D014461:Ultracentrifugation
C506642:CETP protein, human; D002352:Carrier Proteins; D053480:Cholesterol Ester Transfer Proteins; D002788:Cholesterol Esters; D008078:Cholesterol, LDL; D006023:Glycoproteins; D008077:Lipoproteins, LDL; D014316:Tritium