HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Böttcher, A. Articles by Schmitz, G. Search for Related Content PubMed PubMed Citation Articles by Böttcher, A. Articles by Schmitz, G. Social Bookmarking                      What's this?

Journal of Lipid Research, Vol. 41, 905-915, June 2000
Copyright © 2000 by Lipid Research, Inc.

Original Article

Preparative free-solution isotachophoresis for separation of human plasma lipoproteins: apolipoprotein and lipid composition of HDL subfractions

Correspondence to: Gerd Schmitz

We have previously shown that plasma lipoproteins can be separated by analytical capillary isotachophoresis (ITP) according to their electrophoretic mobility in a defined buffer system. As in lipoprotein electrophoresis, HDL show the highest mobility followed by VLDL, IDL, and LDL. Chylomicrons migrate according to their net-charge between HDL and VLDL, because ITP has negligible molecular sieve effects. Three HDL subfractions were obtained which were designated fast-, intermediate-, and slow-migrating HDL. To further characterize these HDL subfractions, a newly developed free-solution ITP (FS-ITP)-system was used, that allows micro-preparative separation of human lipoproteins directly from whole plasma (Böttcher, A. et al. 1998. Electrophoresis. 19: 1110;–1116). The fractions obtained by FS-ITP were analyzed for their lipid and apolipoprotein composition and by two-dimensional nondenaturing polyacrylamide gradient gel electrophoresis (2D-GGE) with subsequent immunoblotting. fHDL are characterized by the highest proportion of esterified cholesterol of all three subfractions and are relatively enriched in LpA-I. Together with iHDL they contain the majority of plasma apoA-I, while sHDL contain the majority of plasma apoA-IV, apoD, apoE, and apoJ. Pre-ß-HDL were found in separate fractions together with triglyceride-rich fractions between sHDL and LDL.

In summary, ITP can separate the bulk of HDL into lipoprotein subfractions, which differ in apolipoprotein composition and electrophoretic mobility. While analytical ITP permits rapid separation and quantitation for diagnostic purposes, FS-ITP can be used to obtain these lipoprotein subfractions on a preparative scale for functional analysis. As FS-ITP is much better suited for preparative purposes than gel electrophoresis, it represents an important novel tool for the functional analysis of lipoprotein subclasses.—Böttcher, A., J. Schlosser, F. Kronenberg, H. Dieplinger, G. Knipping, K. J. Lackner, and G. Schmitz. Preparative free-solution isotachophoresis for separation of human plasma lipoproteins: apolipoprotein and lipid composition of HDL subfractions. J. Lipid Res. 2000. 41: 905;–915.

Supplementary key words: isotachophoresis, apolipoproteins, pre-ß HDL, two-dimensional gel electrophoresis

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				L. Li, P. A. Thompson, and R. L. Kitchens Infection induces a positive acute phase apolipoprotein E response from a negative acute phase gene: role of hepatic LDL receptors J. Lipid Res., August 1, 2008; 49(8): 1782 - 1793. [Abstract] [Full Text] [PDF]

				S. Imaizumi, S.-i. Miura, K. Nakamura, Y. Kiya, Y. Uehara, B. Zhang, Y. Matsuo, H. Urata, M. Ideishi, K.-A. Rye, et al. Antiarrhythmogenic effect of reconstituted high-density lipoprotein against ischemia/reperfusion in rats. J. Am. Coll. Cardiol., April 22, 2008; 51(16): 1604 - 1612. [Abstract] [Full Text] [PDF]

				N. Settasatian, P. J. Barter, and K.-A. Rye Remodeling of apolipoprotein E-containing spherical reconstituted high density lipoproteins by phospholipid transfer protein J. Lipid Res., January 1, 2008; 49(1): 115 - 126. [Abstract] [Full Text] [PDF]

				B. Zhang, Y. Uehara, S. Hida, S.-i. Miura, D. L. Rainwater, M. Segawa, K. Kumagai, K.-A. Rye, and K. Saku Effects of reconstituted HDL on charge-based LDL subfractions as characterized by capillary isotachophoresis J. Lipid Res., May 1, 2007; 48(5): 1175 - 1189. [Abstract] [Full Text] [PDF]

				K.-A. Rye, R. Bright, M. Psaltis, and P. J. Barter Regulation of reconstituted high density lipoprotein structure and remodeling by apolipoprotein E J. Lipid Res., May 1, 2006; 47(5): 1025 - 1036. [Abstract] [Full Text] [PDF]

				B. Zhang, T. Kaneshi, T. Ohta, and K. Saku Relation between insulin resistance and fast-migrating LDL subfraction as characterized by capillary isotachophoresis J. Lipid Res., October 1, 2005; 46(10): 2265 - 2277. [Abstract] [Full Text] [PDF]

				B. Zhang, P. Fan, E. Shimoji, H. Xu, K. Takeuchi, C. Bian, and K. Saku Inhibition of Cholesteryl Ester Transfer Protein Activity by JTT-705 Increases Apolipoprotein E-Containing High-Density Lipoprotein and Favorably Affects the Function and Enzyme Composition of High-Density Lipoprotein in Rabbits Arterioscler. Thromb. Vasc. Biol., October 1, 2004; 24(10): 1910 - 1915. [Abstract] [Full Text] [PDF]

				B. Zhang, A. Matsunaga, K. Saku, S. Nakano, and T. Yamada Associations Among Plasma Lipoprotein Subfractions as Characterized by Analytical Capillary Isotachophoresis, Apolipoprotein E Phenotype, Alzheimer Disease, and Mild Cognitive Impairment Arterioscler. Thromb. Vasc. Biol., August 1, 2004; 24(8): e144 - e144. [Full Text] [PDF]

				B. Ezeh, M. Haiman, H. F. Alber, B. Kunz, B. Paulweber, A. Lingenhel, H.-G. Kraft, F. Weidinger, O. Pachinger, H. Dieplinger, et al. Plasma distribution of apoA-IV in patients with coronary artery disease and healthy controls J. Lipid Res., August 1, 2003; 44(8): 1523 - 1529. [Abstract] [Full Text] [PDF]

				W. Safi, J. N. Maiorano, and W. S. Davidson A proteolytic method for distinguishing between lipid-free and lipid-bound apolipoprotein A-I J. Lipid Res., May 1, 2001; 42(5): 864 - 872. [Abstract] [Full Text]

				S. Barlage, D. Fröhlich, A. Böttcher, M. Jauhiainen, H. P. Müller, F. Noetzel, G. Rothe, C. Schütt, R. P. Linke, K. J. Lackner, et al. ApoE-containing high density lipoproteins and phospholipid transfer protein activity increase in patients with a systemic inflammatory response J. Lipid Res., February 1, 2001; 42(2): 281 - 290. [Abstract] [Full Text]