A low temperature flotation method to rapidly isolate lipoproteins from plasma
| Autor: | Mike VanRollins, Howard R. Knapp, H. Tong |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 1998 |
| Předmět: |
chemistry.chemical_classification
Very low-density lipoprotein Chromatography very low density lipoproteins Triglyceride human plasma Cholesterol Cell Biology QD415-436 Biochemistry chemistry.chemical_compound ultracentrifugation Endocrinology High-density lipoprotein Enzyme chemistry low density lipoproteins Agarose lipids (amino acids peptides and proteins) Ultracentrifuge high density lipoproteins Bradford protein assay mass spectrometry |
| Zdroj: | Journal of Lipid Research, Vol 39, Iss 8, Pp 1696-1704 (1998) |
| ISSN: | 0022-2275 |
| Popis: | To minimize oxidative modification, a low tem- perature, sequential flotation method was developed to isolate plasma lipoproteins in 18 h using a benchtop ultra- centrifuge. The protein distributions were characterized using agarose and SDS-polyacrylamide gel electrophore- sis, and an SDS-Lowr y protein assay. The lipid distribu- tions were assessed using a gas chromatography-mass spec- trometric assay for cholesterol and an enzymatic assay for triglycerides. To validate the rapid flotation method, lipoproteins were also isolated from the same plasma samples using a modified Havel et al. flotation method ( J. Clin. Invest. 34: 1345-1353, 1955). The same lipoproteins and apolipoproteins were present in fractions of compa- rable density, and the summed recoveries of protein, cho- lesterol, and triglyceride were also identical for the Havel et al. and rapid fl otation procedures. Likewise, the amount of cholesterol and triglyceride in corresponding very low, intermediate, and low density lipopr otein (VLDL/IDL and LDL) fractions was the same for the two flotation procedures. The triglyceride and cholesterol lev- els in high density lipoprotein (HDL) isolated by rapid flo- tations, however, were 9-12% higher than in the HDL as isolated by Havel et al. Because a 9 -12% increase in the HDL fraction reflects only 1- 4% of the total triglyceride and cholesterol in plasma, we conclude that, while main- tained at 4 8 C, lipoproteins were quantitatively isolated from human plasma in 1 day.— Tong, H., H. R. Knapp, and M. VanRollins. A low temperature flotation method to rapidly isolate lipoproteins from plasma. J. Lipid. Res. 1998. 39: 1696-1704. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|