Purification of selenoprotein P from human plasma |
| |
| Affiliation: | 1. Division of Gastroenterology, Department of Medicine and Center in Molecular Toxicology, MCN-C2104, Vanderbilt University, Nashville, TN 37232-2279, USA;1. Department of Biochemistry, School of Medicine, Iran University of Medical Sciences, Tehran, Iran;2. Metabolic Disorders Research Center, Endocrinology and Metabolism Research Institute, Tehran University of Medical Sciences, Tehran, Iran;3. Department of Biochemistry, School of Medicine, Yazd University of Medical Sciences, Yazd, Iran;4. H.Aliasghar Hospital, Iran University of Medical Sciences, Tehran, Iran;5. Department of Biochemistry, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran;1. Institute of Nutrition, Department of Nutrigenomics, Friedrich-Schiller-Universität Jena, D-07743 Jena, Germany;2. Faculty of Pharmacy and Pharmaceutical Sciences, University of Alberta, Edmonton, AB, Canada;1. GATA Haydarpasa Training Hospital, Department of Internal Medicine, 34668 Istanbul, Turkey;2. GATA Haydarpasa Training Hospital, Department of Gastroenterology, 34668 Uskudar, Istanbul, Turkey;3. GATA Haydarpasa Training Hospital, Department of Radiology, 34668 Istanbul, Turkey;4. GATA Haydarpasa Training Hospital, Department of Biochemistry, 34668 Istanbul, Turkey;1. Department of Nutrition, Faculty of Public Health, Zabol University of Medical Sciences, Zabol, Iran;2. Department of Clinical Nutrition, School of Nutritional Sciences and Dietetics, Tehran University of Medical Sciences, Tehran, Iran;3. Department of Epidemiology and Biostatistics, Faculty of Public Health, Zabol University of Medical Sciences, Zabol, Iran |
| |
| Abstract: | Selenoprotein P was partially purified (> 1000-fold) from human plasma in four chromatographic steps using 75Se-labeled selenoprotein P secreted by HepG2 cells in culture as a marker. The purified preparation was injected into mice and monoclonal antibodies, which precipitated the labeled protein, were generated. Neither of two different monoclonal antibodies had cross-reactivity with plasma from five animal species. Antibodies were coupled to agarose, and selenoprotein P was purified from human plasma by immunoaffinity chromatography followed by chromatography on heparin agarose. With two different matrix-bound monoclonal antibodies, the purification procedure gave two bands on SDS-PAGE with mobilities corresponding to 61 and 55 kDa. Both bands stained for carbohydrate and showed increased electrophoretic mobility after enzymatic deglycosylation. Immunoaffinity chromatography removed approx. one-third of the selenium from plasma or 0.4 μmol Se/l at a total selenium concentration of 1.1 μmol/l, indicating that selenoprotein P constituted this proportion of total plasma selenium in healthy US blood donors. |
| |
| Keywords: | |
| 本文献已被 ScienceDirect 等数据库收录！ |
|
