scholarly journals Characterization of plasma lipoproteins separated and purified by agarose-column chromatography

1974 ◽  
Vol 139 (1) ◽  
pp. 89-95 ◽  
Author(s):  
Lawrence L. Rudel ◽  
Jason A. Lee ◽  
Manford D. Morris ◽  
James M. Felts

1. A simple method for isolation of individual human plasma lipoprotein classes is presented. In this technique, lipoproteins are removed from plasma at d1.225 by ultracentrifugation, after which they are separated and purified by agarose-column chromatography. 2. Three major classes are obtained after agarose-column chromatography. Separation between classes is excellent; more than 95% of the lipoproteins eluted from the column are recovered in the form of a purified lipoprotein class. 3. Each lipoprotein class was characterized immunologically, chemically, electrophoretically and by electron microscopy. A comparison of the properties of the column-isolated lipoproteins was made with very-low-density lipoproteins, low-density lipoproteins, and high-density lipoproteins separated by sequential ultracentrifugation at densities of 1.006, 1.063 and 1.21 respectively. 4. By each criterion, peak-I lipoproteins from the agarose column are the same as very-low-density lipoproteins, peak-II lipoproteins are the same as low-density lipoproteins, and peak-III lipoproteins are the same as high-density lipoproteins. Thus the lipoprotein classes isolated by both methods are similar if not identical. 5. The agarose-column separation technique offers the advantage of a two- to three-fold saving in time. In addition, the column-elution pattern serves as a recording of the size distribution of lipoproteins in plasma. 6. The most complete characterization is reported for human plasma lipoproteins. The results with rhesus-monkey and rabbit lipoproteins were identical.

1957 ◽  
Vol 105 (1) ◽  
pp. 49-67 ◽  
Author(s):  
Frederick Aladjem ◽  
Miriam Lieberman ◽  
John W. Gofman

Low density human plasma lipoproteins Sf 17+, Sf 13, and Sf 6, high density lipoproteins 2 and 3, and a lipoprotein-free plasma protein fraction were isolated from human plasma by ultracentrifugal methods. It was found that human plasma lipoproteins are immunochemically distinct from the non-lipoprotein containing plasma protein fraction. Lipoprotein fractions of a given hydrated density, isolated from different individuals, were found to be immunochemically indistinguishable by qualitative absorption tests. Qualitative antigenic differences were shown to exist between low density lipoproteins and high density lipoproteins. Quantitative precipitin reactions showed that low density lipoproteins Sf 6 and Sf 13 were immunochemically very similar. However, they differed with respect to the amount of antigen nitrogen required for maximum precipitation. Agar diffusion analyses were performed; the results suggest heterogeneity of lipoproteins by this criterion.


1977 ◽  
Vol 146 (6) ◽  
pp. 1791-1803 ◽  
Author(s):  
J H Morse ◽  
L D Witte ◽  
D S Goodman

Lipoproteins, isolated by sequential flotation at densities 1.006, 1.019, 1.063, and 1.21, were examined for their ability to inhibit human lymphocytes stimulated by allogeneic cells and by lectins (phytohemagglutinin-P and concanavalin A). All the classes of normal plasma lipoproteins inhibited lymphoproliferation when peripheral blood lymphocytes were cultured in autologous, heterologous, or lipoprotein-deficient plasma (d greater than 1.21). The rank order of inhibitory potency was intermediate density lipoprotein (IDL) greater than very low density lipoproteins (VLDL) greater than low density lipoproteins (LDL) greater than high density lipoproteins (HDL), regardless of the mode of stimulation. The concentrations of IDL, VLDL, and LDL required for complete inhibition of stimulated lymphoproliferation were considerably below the levels of each of these lipoproteins normally found in human plasma. In addition, the concentration of HDL required for 50-90% inhibition was in the range of HDL levels normally found in human plasma. Moreover, at relatively higher concentrations, lipoproteins suppressed the incorporation of [3H]thymidine into DNA below the levels seen with reseting, unstimulated lymphocytes. The results suggest that circulating lymphocytes may normally be highly suppressed by the combined effects of all the endogenous lipoproteins and that the lipoproteins may play important roles in vivo in modulating lymphocyte functions and responses.


1986 ◽  
Vol 32 (2) ◽  
pp. 283-286 ◽  
Author(s):  
J E Groener ◽  
R W Pelton ◽  
G M Kostner

Abstract This simple, routine assay for measuring cholesteryl ester transfer/exchange activity in human plasma is based on the removal of interfering lipoproteins--very-low-density (VLDL) and low-density lipoproteins (LDL)--by precipitation with polyethylene glycol. High-density lipoproteins (HDL) in the samples do not affect the results. The supernate after precipitation is mixed with [14C]cholesteryl ester-labeled LDL as donor and with HDL as the acceptor for the cholesteryl ester. After incubation for 16 h at 37 degrees C, LDL is separated from HDL by precipitation with dextran sulfate and the radioactivity measured in the supernate, which contains the HDL. The assay is applicable to samples containing as much as 10 mmol of triglycerides per liter. The within-assay CV was 2.7%, the day-to-day CV 6.8%. Results compared well with those by conventional procedures.


1979 ◽  
Vol 25 (10) ◽  
pp. 1795-1798 ◽  
Author(s):  
I R Kupke ◽  
S Zeugner ◽  
A Gottschalk

Abstract We compared the results obtained by a micromethod for the determination of plasma lipoprotein cholesterol, in which electrophoresis is used to separate the lipoprotein fractions (beta-, pre-beta-, and alpha-lipoproteins), with those determinations with ultracentrifugation (low-density, very-low-density, and high-density lipoproteins). Precision of determination (coefficient of variation, CV, %) was the same for beta- and low-density lipoproteins (1.6%), and for pre-beta- and very-low-density lipoproteins (3.7%); however, determination of alpha-lipoprotein cholesterol was more precise (1.4%) than that of high-density lipoprotein cholesterol (3.1%). Analytical recovery of lipoprotein cholesterol was the same for both methods (98--100%) and the results were closely correlated (r = 0.943). The procedure has been used to determine the cholesterol content of plasma lipoprotein fractions of apparently healthy adults (both sexes). Lipoprotein cholesterol concentrations in our population sample compare well with those reported for other groups of similar age, in particular Stanford long-distance runners.


Author(s):  
Sawsan Taha Ahmed al-Haddad ◽  
Zaid Mohammed Mubarak Almahdawi ◽  
Munife S. Ahmed Al-janabi

This study was designed to test the therapeutic efficacy of some hypotensive drugs and vegetable drinks on some biochemical indicators in male rabbits, where atherosclerosis was developed using 1% cholesterol with food. This study was conducted in June until the end of July 2017 in the Pharmacology Department/ General Company for Pharmaceutical Industry in Samarra. In the study, 50 local rabbits were randomly distributed by 10 groups each containing 5 animals. The first group considered as the control group. The second group is the control group treated with 1% cholesterol with the food, the third group treated with cholesterol (1% and captopril 0.71 mg), group 4 (cholesterol 1% with atenolol 0.71 mg / kg), group 5 (cholesterol 1%, amlodipine 0.07 mg / kg) , group 6 treated with cholesterol 1% and aldomet (0.57 mg / kg), group 7 (cholesterol 1% and furosemide at 3.5 mg / kg), group 8 (cholesterol 1% with garlic syrup 2 ml), group 9 treatment cholesterol 1% and lemon juice), and group 10 Treatment with (1% cholesterol and green tea syrup 2 ml). The results of the study showed a significant increase (P≤0.01)) at the level of each of cholesterol triple and triglycerides, proteins and low density lipoproteins, very low density lipoproteins, also led to obtain a significant decrease in the level of high-density lipoproteins (HDL) in the treatment group with cholesterol 1% compared to control group. At the time of the treatment of anti- pressure drugs: Captopril, Atenolol, Amlodipine ,Aldomet, and Furosmide , there were no significant differences in the cholesterol level of all pharmacological groups. Moral differences were not found in LDL-C and there was a significant decrease (P≤0.01) of the level of triglycerides, proteins and very low density lipoproteins, and there was a significant increase in the level of high-density lipoproteins HDL-C, while treatment with plant juices, there was a significant decrease (P≤0.01) in the level of total cholesterol and triglycerides and LDL, and VLDL, high-density lipoprotein (HDL-C) increased when treated with garlic, lemon and green tea. We conclude pressure drugs of any kind can cure atherosclerosis or prevent high fat, unlike its counterparts OF plants, which have shown a significant effect on controlling lipid profile and reducing their effects and future risks on the heart.


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