SUBFRACTIONATION OF THE Sf 20-105 LIPOPROTEINS IN A SWINGING BUCKET ROTOR1

Lindgren, F. T.; Nichols, Alex V.; Upham, Frank T.; Wills, R. D.

doi:10.1021/j100816a042

Cited by 34 publications

(9 citation statements)

References 8 publications

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“…After removal of the top 0.5 ml, the density of the VLDL solution was increased to 1.065 and it was placed in A% X 31-inch thickwalled cellulose nitrate tubes (Beckman Instruments, Inc., Spinco Div.). The gradient was layered as described by Lindgren et al (22). The tubes were centrifuged at 35,000 rpm in a Beckman SW 41 rotor.…”

Section: Methodsmentioning

confidence: 99%

Apoprotein composition of very low density lipoproteins of human serum.

Kane¹,

Sata²,

Hamilton³

et al. 1975

J. Clin. Invest.

369

107

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A B S T R Asize from normolipidemic and hyperlipemic subjects with the exception that all fractions from the hyperlipemic subjects contained more R-serine and some, more arginine rich polypeptide. Even in the absence of chylomicrons, the distribution of soluble apoproteins in particles of diameters greater than 700-800 A was usually similar to that of the smallest particles. This suggests that the largest particles may include products of the partial catabolism of chylomicrons. INTRODUCTIONThe protein moiety of very low density lipoproteins (VLDL)1 of human serum contains several discrete polypeptides. One of the predominant species, apolipoprotein B (apoB), which appears to be identical with the major apoprotein of low density lipoprotein (LDL) (1), is insoluble in aqueous buffers after delipidation. Several water-soluble polypeptides denoted by their carboxyl termini as the "R-serine," "R-glutamic acid," and "R-alanine" species (2-4) and a major apoprotein, rich in arginine (5) (8) and provides the basis for determination of apoB content by selective precipitation. Under carefully controlled conditions the soluble apoproteins, after delipidation by TMU, migrate quantitatively within reproducible Rf zones upon electrophoresis in polyacrylamide gels. Conditions of staining and destaining have been found which provide linearity of photometric response over a wide range for all the major soluble apoproteins. Determination of the absolute chromogenicity of the purified apoproteins permits quantitation of each in the electrophoretic gels. The distribution of the major apoproteins is presented for whole human serum VLDL from normolipidemic subjects and for fractions of VLDL of different particle diameters. METHODS MaterialsReagents were as described previously (8) except that TMU was redistilled monthly in glass and stored under nitrogen in the dark at 3°C. TMU which is suitably purified has a pH of 6.0-7.0 when diluted fivefold wih water.Amidoschwarz lOB, lot no. 0766421, was from the Allied Chemical Corp., Morristown, N. J. VLDL (d < 1.006) and LDL (1.024 < d < 1.050) were prepared from serum obtained from subjects who had fasted for at least 14 h by repetitive ultracentrifugation as described previously (8). All lipoprotein solutions contained EDTA (1 mM) and sodium azide (0.2 mg/ml).All normolipidemic donors were healthy students and hospital personnel between 20 and 60 yr of age, with serum levels of cholesterol and triglyceride below 250 and 140 mg/dl, respectively. To avoid preselection bias, blood was drawn for the preparation of whole VLDL and VLDL subfractions from putative normal donors without pretesting. Preparations from all subjects with serum lipid levels below the limits given above were then studied. Subjects with hyperlipemia were free of known causes of secondary hyperlipidemia or serious disease other than atherosclerotic vascular disease or gout. Serum was examined for chylomicrons after standing overnight at 3°C (9) and by electrophoresis in agarose gel (10). Samples containing detectible chylo...

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Section: Methodsmentioning

confidence: 99%

Apoprotein composition of very low density lipoproteins of human serum.

Kane¹,

Sata²,

Hamilton³

et al. 1975

J. Clin. Invest.

369

107

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“…Nevertheless, it is the only method by which one can isolate the individual LP components selectively from serum on a preparative scale. In addition, it is possible to obtain a further sub-fractionation of the LP (e.g., LDL or HDL) by way of density gradients [27,28] .…”

Section: Nmr In Comparison To Other Methodsmentioning

confidence: 99%

NMR spectroscopy – a modern analytical tool for serum analytics of lipoproteins and metabolites

Baumstark¹,

Eiglsperger²,

Pfahlert³

et al. 2015

LaboratoriumsMedizin

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NMR spectroscopy is a modern analytical method which is extremely suitable for the analysis of various body fluids. In addition to small metabolite quantification, the method is capable of differentiated lipoprotein subfraction measurements. The technique can be well standardized and allows extensive automation and good sample throughput compared with lipoprotein fractionation via ultracentrifugation. Because all evaluated parameters are determined simultaneously in one measurement, this method is especially suitable for metabolomics approaches in which even subtle changes in metabolite ratios may be relevant. This review gives an overview of the current methods of NMR spectroscopy, analysis strategies, and practical applications in various studies concerning lipid analysis as well as metabolomics. Over the course of the past years, NMR spectroscopy has heavily evolved from a mere research method into a tool that can be expected to play an important role in routine diagnostic testing in the future.

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“…Außerdem ist es möglich, über Dichtegradienten eine weitere Subfraktionierung der LP (z.B. LDL oder HDL) zu erhalten [27,28].…”

Section: Die Nmr Im Vergleich Zu Anderen Methodenunclassified

NMR-Spektroskopie – ein modernes Werkzeug zur Serum-Analytik von Lipoproteinen und Metaboliten

Baumstark¹,

Eiglsperger²,

Pfahlert³

et al. 2014

LaboratoriumsMedizin

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NMR spectroscopy -a modern analytical tool for serum analytics of lipoproteins and metabolites Zusammenfassung: Die NMR-Spektroskopie ist eine moderne analytische Methode, die sich gut zur Analyse unterschiedlichster Körperflüssigkeiten eignet. Im Serum können neben kleinen Metaboliten auch differenziert Lipoprotein-Fraktionen bestimmt werden. Die Technik ist gut standardisierbar und erlaubt im Vergleich zur Lipoproteinfraktionierung mittels Ultrazentrifugation eine umfassende Automatisierung und guten Probendurchsatz. Da in einer Messung alle auszuwertenden Parameter gleichzeitig gemessen werden, eignet sich die Methode besonders gut für Metabolomics-Ansätze, in denen auch subtile Verschiebungen in den Metabolitenverhältnissen von Bedeutung sein können. Dieser Review gibt eine Übersicht über aktuelle Ansätze der NMR-spektroskopischen Messtechnik, Auswertestrategien und praktischen Anwendungen in verschiedenen Studien, sowohl auf der Seite der Lipidanalyse, also auch hinsichtlich des Metabolitenprofils. Die NMR-Spektroskopie hat sich in den letzten Jahren von einer reinen Forschungsmethode mit großen Schritten weiterentwickelt und dürfte in Zukunft einen Platz in der Routinediagnostik einnehmen.Abstract: NMR spectroscopy is a modern analytical method which is extremely suitable for the analysis of various body fluids. In addition to small metabolite quantification, the method is capable of differentiated lipoprotein subfraction measurements. The technique can be well standardized and allows extensive automation and good sample throughput compared with lipoprotein fractionation via ultracentrifugation. Because all evaluated parameters are determined simultaneously in one measurement, this method is especially suitable for metabolomics approaches in which even subtle changes in metabolite ratios may be relevant. This review gives an overview of the current methods of NMR spectroscopy, analysis strategies, and practical applications in various studies concerning lipid analysis as well as metabolomics. Over the course of the past years, NMR spectroscopy has heavily evolved from a mere research method into a tool that can be expected to play an important role in routine diagnostic testing in the future.

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SUBFRACTIONATION OF THE S_f 20-10⁵ LIPOPROTEINS IN A SWINGING BUCKET ROTOR¹

Cited by 34 publications

References 8 publications

Apoprotein composition of very low density lipoproteins of human serum.

Apoprotein composition of very low density lipoproteins of human serum.

NMR spectroscopy – a modern analytical tool for serum analytics of lipoproteins and metabolites

NMR-Spektroskopie – ein modernes Werkzeug zur Serum-Analytik von Lipoproteinen und Metaboliten

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