Apolipoprotein C-1 (apo C-1) enriched HDL has been described as an atherogenic form
of HDL associated with an increased risk for cardiovascular disease (CVD). The
objective of the present study was to develop a rapid method for the separation,
purification, and characterization of Apo C-1 from serum. We isolated and characterize
HDL subclasses from individuals with and without angiographically-proven CVD who
have elevated and normal-to-low HDL-C levels.
Ultracentrifugation was linked with immunoaffinity separations for the specific
separation of Apo C-1 enriched HDL from other lipoproteins. A 50 μL sample of serum
is diluted in TRIS HCl buffer (pH 7.5) and incubated with CNBr-activated Sepharose
(Amersham) containing antibodies to apo C-1 (Academy Bio-medical Company). The
apo C-1-depleted serum is removed by centrifugation and all apo C-1-containing
lipoproteins are released from the Sepharose beads at pH 2. The apo C-1-depleted
sample and the apo C-1-containing sample were ultracentrifuged to obtain a lipoprotein
density profile in the absence and presence of apo C-1. Density Lipoprotein Profiling
(DLP) gives relevant information of lipoproteins, such as density and subclass characterization, and is a novel approach to purify apo C-1-enriched HDL. An
additional advantage of this approach is that lipoprotein-a (Lp(a)), which is often an
interfering component in the HDL density region, is eliminated.
Results show feasibility that these methods could be used in a clinical setting,
was achieved. This measurement may yield a precise and quantitative profile of the
distribution of apo C-1 for all lipoprotein particles including HDL.
| Identifer | oai:union.ndltd.org:tamu.edu/oai:repository.tamu.edu:1969.1/ETD-TAMU-3123 |
| Date | 15 May 2009 |
| Creators | Lester, Sandy Marie |
| Contributors | Macfarlane, Ronald D. |
| Source Sets | Texas A and M University |
| Language | en_US |
| Detected Language | English |
| Type | Book, Thesis, Electronic Thesis, text |
| Format | electronic, application/pdf, born digital |
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