Quantitation of baboon lipoproteins by high performance gel exclusion chromatography |
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Authors: | Mary C Williams Rampratap S Kushwaha Henry C McGill Jr |
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Affiliation: | (1) Department of Physiology and Medicine, Southwest Foundation for Biomedical Research, P.O. Box 28147, 78284 San Antonio, TX;(2) Department of Pathology, University of Texas Health Science Center, San Antonio, Texas |
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Abstract: | High performance liquid chromatography with gel exclusion columns was used for quantitative measurement of plasma lipoproteins.
A combination of columns TKS 4000 PW and 3000 PW gave good separation of very low (VLDL), low (LDL) and high (HDL) density
lipoproteins. The area under each lipoprotein peak detected by absorbance at 280 nm was measured by digitizing and was expressed
as cm2. Purified lipoprotein standards isolated by ultracentrifugation were also chromatographed in increasing concentrations. The
area under the lipoprotein standard peak was linearly related to the amount of total protein over a wide range. The areas
of most of the measured plasma lipoproteins were within the linear range. The relationship between the area and the amount
of protein for each standard was used to quantitate the amount of protein and was expressed as mg/dl plasma. This technique
is simple and requires a small amount of plasma. The validated technique was applied to a large population of pedigreed baboons.
An average plasma lipoprotein profile of feral baboons on the chow diet was characterized by a high level of HDL (90.9±30.7
mg/dl) with a lesser amount of LDL (29.1±13.2 mg/dl). VLDL was present in much lower concentration (8.6±2.6 mg/dl). Feeding
a high cholesterol and high saturated fat (HCHF) diet raised both LDL (1.5-fold) and HDL levels (1.3-fold) without changing
VLDL levels. Progeny of sires with low response to dietary cholesterol increased their HDL protein when challenged with HCHF
diet without any change in their LDL or VLDL. Progeny of high-responding sires, however, had increases in both their HDL and
LDL levels when challenged with HCHF diet. The survey of lipoprotein profiles of the pedigreed baboon colony disclosed a number
of animals with interesting and unusual lipoprotein patterns. |
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