Rapid Isolation of Human Plasma Lipoproteins
Document Type
Article
Publication Date
12-1-1997
Description
For many studies it is important to isolate lipoproteins quickly from plasma. Current methods require a table top ultracentrifuge with high speed rotors 100-120,000 rpm. The objective of these studies was to develop a method for rapid isolation of plasma lipoproteins using a fixed angle rotor and a full-size ultracentrifuge which are commonly found in biomedical laboratories. We used a Beckman L5 70 ultracentrifuge, a type 80 Ti rotor and the Beckman g-Max system using 4.2 mL quick seal tubes. Two mL of fasting human plasma were placed in each tube and filled with a KBr solution d= 1.006 g/mL and heat sealed. These tubes were then centrifuged at 14° C for 2 hours at 68,000 rpm. The floating very low density lipoproteins (VLDL) were removed using a Beckman tube slicer. The dense bottom portion was adjusted to d= 1.063 g/mL by adding solid KBr and then centrifuged at 14° C for 2.5 hours at 68,000 rpm. The floating low density lipoproteins (LDL) were recovered at the top of the tube by slicing the tube. The bottom portion containing high density lipoproteins (HDL) and other serum proteins was also recovered. Control lipoproteins were isolated by the standard sequential ultracentrifugation at 14° C for 22 hours at 40,000 rpm using the Type 80 Ti rotor. Purity of the fractions was verified by lipid staining after agarose gel electrophoresis. Purity and electrophoretic mobility of lipoproteins isolated by the rapid procedure were identical to those of the standard two day sequential procedure. This new procedure offers rapid separation of plasma lipoproteins using equipment that is routinely available.
Citation Information
Kelley, J. L.; Assadourian, A.; Krishnaswamy, G.; and Chi, D.. 1997. Rapid Isolation of Human Plasma Lipoproteins. FASEB Journal. Vol.11(3). ISSN: 0892-6638