Boettcher, A. and Schlosser, J. and Kronenberg, F. and Dieplinger, H. and Knipping, G. and Lackner, K. J. and Schmitz, Gerd (2000) Preparative free-solution isotachophoresis for separation of human plasma lipoproteins: apolipoprotein and lipid composition of HDL subfractions. JOURNAL OF LIPID RESEARCH, 41 (6). pp. 905-915. ISSN 0022-2275, 1539-7262
Full text not available from this repository. (Request a copy)Abstract
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, LDL, 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 front whole plasma (Bottcher, 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-beta-HDL were found in separate fractions together with triglyceride-rich fractions between sHDL and LDL, jlr 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.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | HIGH-DENSITY-LIPOPROTEIN; ANALYTICAL CAPILLARY ISOTACHOPHORESIS; FREE-FLOW ISOTACHOPHORESIS; CELL-DERIVED CHOLESTEROL; A-I; FACTOR-H; GEL-ELECTROPHORESIS; PROTEINS; SUBCLASSES; CHROMATOGRAPHY; isotachophoresis; apolipoproteins; pre-beta HDL; two-dimensional gel electrophoresis |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Klinische Chemie und Laboratoriumsmedizin |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 26 Apr 2022 14:33 |
| Last Modified: | 26 Apr 2022 14:33 |
| URI: | https://pred.uni-regensburg.de/id/eprint/42444 |
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