Rapid method for the isolation of lipoproteins from human serum by precipitation with polyanions
TL;DR: With a single preparative ultracentrifugation, immunologically pure high density lipoproteins can be isolated from large volumes of serum.
About: This article is published in Journal of Lipid Research.The article was published on 1970-11-01 and is currently open access. It has received 2449 citations till now.
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TL;DR: A simplified method for measuring high-density lipoprotein cholesterol in serum after very-low- and low- density lipoproteins have been precipitated from the specimen with sodium phosphotungstate and Mg2+.
Abstract: We describe a simplified method for measuring high-density lipoprotein cholesterol in serum after very-low- and low-density lipoproteins have been precipitated from the specimen with sodium phosphotungstate and Mg2+. Values so obtained correlate well with values obtained with the heparin-Mn2+ precipitation technique (r = 0.95, CV less than 5% in 66% of the subjects studied and between 5 and 10% in the remaining ones) or by ultracentrifugal separation (r = 0.82, CV less than 5% in 80% of the subjects studied and between 5 and 10% in the remaining ones). Our precipitation technique is more appropriate for routine clinical laboratory use.
1,802 citations
Cites result from "Rapid method for the isolation of l..."
...We find the last method (8) to be simple, reliable, precise, and inexpensive and that it gives results comparable to those obtained by older, more cumbersome methods....
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...Here we report the results of a study in which HDL cholesterol was determined by these three different methods (4, 5,8)....
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TL;DR: Determination of the extent of the unprecipitated apoB-associated lipoproteins by sensitive radioimmunoassay and of the amount of precipitated high density lipoprotein by radial immunodiffusion assay of apolipoproteins A-I and A-II at various heparin and Mn(2+) concentrations indicated that the usual Heparin level was adequate.
1,297 citations
TL;DR: The relationship of future clinical coronary heart-disease to the plasma-high-density-lipoprotein (H.H.D.L.)-cholesterol concentration has been examined in a 2-year case-control follow-up study of 6595 men aged 20-49 years living in the municipality of Tromsø, Norway and support the proposal that a low H.d.L. cholesterol concentration is important in accelerating the progression of coronary atherosclerosis.
1,180 citations
TL;DR: In this article, the relation between the fatty acid composition of skeletal-muscle phospholipids and insulin sensitivity was determined in two groups of subjects, including 27 patients undergoing coronary artery surgery and 13 normal men.
Abstract: Background Insulin resistance and hyperinsulinemia are features of obesity, non-insulin-dependent diabetes mellitus, and other disorders. Skeletal muscle is a major site of insulin action, and insulin sensitivity may be related to the fatty-acid composition of the phospholipids within the muscle membranes involved in the action of insulin. Methods We determined the relation between the fatty-acid composition of skeletal-muscle phospholipids and insulin sensitivity in two groups of subjects. In one study, we obtained samples of the rectus abdominis muscle from 27 patients undergoing coronary artery surgery; fasting serum insulin levels provided an index of insulin sensitivity. In the second study, a biopsy of the vastus lateralis muscle was performed in 13 normal men, and insulin sensitivity was assessed by euglycemic-clamp studies. Results In the patients undergoing surgery, the fasting serum insulin concentration (a measure of insulin resistance) was negatively correlated with the percentage of individua...
834 citations
TL;DR: A simple precipitation method for the separation of HDL subclasses in human plasma found that HDL2 is the more variable component and may be a more meaningful index of altered HDL metabolism.
805 citations
References
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TL;DR: An investigation of the biochemical changes following experimental liver injury felt the need of a simple, rapid, and accurate method for determining the protein fractions in small amounts of serum and began with Kingsley’s biuret procedure.
15,717 citations
TL;DR: The relatively low density of the lipoproteins was utilized by Lindgren, Elliott, and Gofman to separate them from the other serum proteins by ultracentrifugal flotation, and quantitation was subsequently performed by refractometric methods in the analytical ultracentRifuge.
Abstract: In the past few years several methods have been developed for the analysis of serum lipoproteins Lindgren, Elliott, and Gofman (1) have utilized the relatively low density of the lipoproteins to separate them from the other serum proteins by ultracentrifugal flotation Quantitation was subsequently performed by refractometric methods in the analytical ultracentrifuge Separations of lipoproteins have also been made by Cohn fractionation in cold ethanol, and the quantities of lipoprotein have been estimated from the lipid content of the fractions (2, 3) Widely used at the present time is the method of zone electrophoresis with quantitation either by staining (4) or by chemical analysis of eluates from the support
8,544 citations
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2,543 citations
TL;DR: Working on the separated and eluted fractions of the toxin of Corynebacterium diphtheriae in tissue cultures and in animals indicated that the resolution of these proteins was not completely satisfactory in the buffer.
Abstract: THE recently developed starch gel electrophoresis has proved to have a resolving power superior to any other type of zone electrophoresis1–4. I have applied this technique to the study of highly purified toxins3, but further work on the separated and eluted fractions of the toxin of Corynebacterium diphtheriae in tissue cultures and in animals indicated that the resolution of these proteins was not completely satisfactory in the buffer (0.03 M boric acid and 0.012 M sodium hydroxide) normally used in this type of zone electrophoresis. A more suitable system was found with a mixture of tris-(hydroxymethyl)-aminomethane-citrate (tris-citrate) used in conjunction with a boric acid buffer. Preliminary results obtained with toxins, enzymes and human sera are briefly reported here.
1,645 citations