Rapid method for the isolation of two purified subfractions of high density lipoproteins by differential dextran sulfate-magnesium chloride precipitation.
We describe a rapid and reliable three-step precipitation procedure for the isolation of large amounts of the two major components of high density lipoproteins (HDL) in human serum. Precipitation was accomplished by means of dextran sulfate (DS) of mol. wt. 500,000 and MgCl2. First, all apoB-associated lipoproteins of any density were selectively precipitated with critical concentrations of reagents. Secondly, a subfraction of HDL was differentially precipitated from the apoB-depleted serum by increasing the concentration of both reagents. Eventually, the bulk of the remainder of HDL was precipitated by lowering the pH to 5.4. According to the precipitation patterns and the density profiles, the DS-Mg procedure provides a clear differentiation between the two HDL components. According to the compositional criteria and the ultracentrifugal characteristics, the two polyanion-precipitated subclasses are very similar to, if not identical with, the two density subclasses, the lighter HDL2 and the heavier HDL3, isolated by preparative ultracentrifugation after apoB-containing lipoproteins had been removed.
Burstein
M
M
Institut National de Transfusion Sanguine, Paris, France.
Fine
A
A
Atger
V
V
Wirbel
E
E
Girard-Globa
A
A
D011232:Chemical Precipitation; D016264:Dextran Sulfate; D003911:Dextrans; D004586:Electrophoresis / Q000379:methods; D005260:Female; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008075:Lipoproteins, HDL / Q000097:blood / Q000302:isolation & purification*; D053439:Lipoproteins, HDL2; D053440:Lipoproteins, HDL3; D008274:Magnesium; D015636:Magnesium Chloride; D008297:Male; D014461:Ultracentrifugation
D003911:Dextrans; D008075:Lipoproteins, HDL; D053439:Lipoproteins, HDL2; D053440:Lipoproteins, HDL3; D015636:Magnesium Chloride; D016264:Dextran Sulfate; D008274:Magnesium