VLDL Apolipoprotein B Determination in Blood Serum Following Precipitation of LDL with Polyvinylsulphate

Schriewer, H; Nolte, W.; Assmann, Gerd

doi:10.1515/cclm.1985.23.6.349

Cited by 6 publications

(4 citation statements)

References 9 publications

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“…HDL cholesterol (HDL-C) was measured after precipitation of very-low-density and low-density lipoproteins with phosphotungstic acid [29]. LDL cholesterol (LDL-C) assay was performed using polyvinylsulphate (Boehr-inger Mannheim) [30]. The apolipoprotein content of lipoproteins was examined by electrophoresis in SDSpolyacrylamide gel (SDS-PAGE) [25].…”

Section: Chemical Analysesmentioning

confidence: 99%

In vitro remodelling of plasma lipoproteins in whole plasma by lipoprotein lipase in primary and secondary hypertriglyceridaemia

Lévy

Deckelbaum

Thibault

et al. 1990

Eur J Clin Investigation

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In patients with familial lipoprotein lipase deficiency (FLPL-d) and glycogen storage disease type I (GSD-I), hypertriglyceridaemia (1445 +/- 247 and 1082 +/- 312 mg dl-1, n = 5 per group) was associated primarily with reduced extrahepatic lipoprotein lipase (LPL) activity (0.33 +/- 0.33 and 1.69 +/- 0.38 mumol FFA ml-1 h-1) when compared with controls (4.83 +/- 0.90). Hypercholesterolaemia was characterized by elevated LDL cholesterol (191 +/- 30 and 344 +/- 34 vs. 115 +/- 5 mg dl-1 in controls P less than 0.01) and low HDL cholesterol (12 +/- 2 and 22 +/- 2 vs. 56 +/- 3 in controls, P less than 0.001). In order to ascertain the role of LPL in the interconversion and remodelling of lipoproteins in these disorders, we analysed lipid and lipoprotein profiles before and following in vitro incubation of patient plasma with purified milk LPL (EC 3.1.1.34) for 6 h at 37 degrees C. The efficiency of exogenous LPL in vitro was demonstrated by the extent of hydrolysis of chylomicrons and of VLDL-TG in both groups. Concomitant with the disappearance of TG-rich lipoprotein particles, a consistent per cent increment of IDL (99.2 +/- 30.8 and 43.9 +/- 70.5), LDL (152.8 +/- 36.2 and 137.0 +/- 36.1) and of HDL2 (144.8 +/- 29.4 and 99.8 +/- 18.7) was observed in both groups of patients. The enhancement of the latter fractions contrasted with the decline of HDL3 mass concentration (25.4 +/- 7.7 and 51.4 +/- 5.8%), suggesting that a major shift of HDL3----HDL2 occurs following in vitro lipolysis by LDL. Simultaneous compositional and morphological changes of individual lipoprotein particles were noted, confirming the dynamic movement and exchange of neutral lipids and proteins. Specificity of LPL results was demonstrated by experiments in which incubation of the whole plasma at 37 degrees C without exogenous lipolytic enzyme did not cause any substantial changes. The present study, therefore, demonstrates a correction of the major lipoprotein abnormalities associated with FLPL-d and GSD-I by exogenous LPL. No substantial difference was noted between primary (FLPL-d) and secondary (GSD-I) hyperlipidaemias. These studies allow us to conclude that a simple in vitro system, utilizing an exogenous source of LPL and plasma from patients, may serve as a suitable model for the study of the metabolic relationships of lipoproteins. However, in view of the fact that the extent of lipolysis achieved in vitro did not differ between FLPL-d and GSD-I, it may not be able to separate primary from secondary hyperlipaemias.

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Section: Chemical Analysesmentioning

confidence: 99%

In vitro remodelling of plasma lipoproteins in whole plasma by lipoprotein lipase in primary and secondary hypertriglyceridaemia

Lévy

Deckelbaum

Thibault

et al. 1990

Eur J Clin Investigation

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“…Radial immu-nodiffusion is calibrated with an apolipoprotein B value obtained by determination of tetramethyl ureainsoluble proteins in the supernatant after polyvinylsulphate precipitation (32). Radial immu-nodiffusion is calibrated with an apolipoprotein B value obtained by determination of tetramethyl ureainsoluble proteins in the supernatant after polyvinylsulphate precipitation (32).…”

Section: Discussionmentioning

confidence: 99%

Determination of Apolipoprotein B in Apolipoprotein CII/CIII-Containing Lipoproteins by an Immunoenzymmetric Assay

Sandkamp¹,

Tambyrajah²,

Assmann³

et al. 1992

Clinical Chemistry and Laboratory Medicine

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A solid phase sandwich immunoenzymmetric assay is described for the determination of apolipoprotein B in apolipoprotein CII-and/or CHI-containing lipoproteins (chylomicron remnants, VLDL, IDL). During a first incubation step the particles containing apolipoproteins CII and/or CIII are bound to antibodies against these components, while the antibodies are immobilised on microtitre plate wells. After washing, antiapolipoprotein B is added in a second incubation step. Finally peroxidase-labelled anti-sheep IgG reacts with the bound anti-apolipoprotein B, thus allowing the quantification of complexes containing both B and CII/ CIII apolipoproteins.The amounts of these complexes were correlated with total cholesterol, triacylglycerols, HDL-and LDLcholesterol, apolipoproteins AI and B, as well as with age and sex of the subject. A total of 258 individuals was studied, including patients with lipid metabolism disorders, patients with manifest coronary heart disease, and healthy controls.The assay described in this article was compared with radial immunodiffusion after pretreatment of samples. The immunoenzymmetric assay was easier and faster to perform and had a lower detection limit than the classical VLDL apolipoprotein B determination using ultracentrifugation. Furthermore, it could be performed directly on native serum.Most importantly, the study revealed elevated apolipoprotein CII/CIII-B levels in coronary heart disease patients of both sexes compared with normal subjects. Furthermore, in male coronary heart disease patients a negative correlation was found between the concentrations of apolipoprotein CII/CIII-B and HDLcholesterol. These results suggest a delayed VLDL-HDL exchange of lipids and proteins in these patients which results in an accumulation of atherogenic apolipoprotein B-containing VLDL and IDL. ') Funding organisation: Ministerium für Wissenschaft, For-altered composition of these particles in patients when schung und Technik Nordrhein-Westfalen, IV B 5-40001986 compared with healthy controls (20, 21). Hence it isEur.

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“…124974 (all Boehringer Mannheim, Germany). Apolipoprotein A-I (apo A-I) was determined in HDL and apolipoprotein B (apo B) in VLDL and LDL by nephelometry [9], VLDL-apo B determination was done after lipase treatment (1 U/Ìl) of triglycerides in the sample [10]. Antisera and standards were purchased from Immuno (Heidelberg, Germany) and Behringwerke (Marburg, Germany).…”

Section: Methodsmentioning

confidence: 99%

Influence of Different Fat Emulsions with 10 or 20% MCT/LCT or LCT on Lipoproteins in Plasma of Patients after Abdominal Surgery

Hailer

Jauch

Wolfram³

1998

Ann Nutr Metab

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In patients after elective abdominal surgery, different fat emulsions were used to compare their efficacy in total parenteral nutrition and in normalizing plasma lipoprotein levels. In five different groups with 5 patients each, half of the nonprotein calories were given as medium-chain triglycerides/long-chain triglycerides (1:1) or as long-chain triglycerides alone in 10 or 20% fat emulsions or as glucose alone in a control group for 7 days. After surgery, an initial decrease of all plasma lipoprotein components was followed by a different behavior of glyceride-glycerol, cholesterol, phospholipids, and apolipoproteins. Glyceride-glycerol in very-low-density lipoproteins and high-density lipoproteins is increasing during infusion of fat emulsions and decreasing during overnight interruption of infusions. After the 7-day infusion period, there was no significant difference in very-low-density lipoprotein glyceride-glycerol as compared with the values before different infusions. Low-density lipoprotein cholesterol is reaching and exceeding preoperative concentrations between the 4th and the 7th day, most during infusion of 10% fat emulsion and especially due to an increase of free cholesterol. High-density lipoprotein cholesterol and apolipoprotein A-I reach preoperative levels during infusion of fat emulsions but not with glucose alone. Higher than preoperative values are reached in phospholipids with all fat infusions already on day 4. Abnormal lipoprotein X occurred least with the medium- chain/long-chain triglyceride 20% fat-infusion. This fat emulsion is suggested as having the best normalizing effect on plasma lipoproteins and best tolerance in patients after surgery.

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VLDL Apolipoprotein B Determination in Blood Serum Following Precipitation of LDL with Polyvinylsulphate

Cited by 6 publications

References 9 publications

In vitro remodelling of plasma lipoproteins in whole plasma by lipoprotein lipase in primary and secondary hypertriglyceridaemia

In vitro remodelling of plasma lipoproteins in whole plasma by lipoprotein lipase in primary and secondary hypertriglyceridaemia

Determination of Apolipoprotein B in Apolipoprotein CII/CIII-Containing Lipoproteins by an Immunoenzymmetric Assay

Influence of Different Fat Emulsions with 10 or 20% MCT/LCT or LCT on Lipoproteins in Plasma of Patients after Abdominal Surgery

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