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A method for purifying the platelet membrane glycoprotein IIb-IIIa complex
Authors:L A Fitzgerald  B Leung  D R Phillips
Affiliation:1. Department of Biomedical Engineering, Faculty of Engineering, University of Malaya, Kuala Lumpur 50603, Malaysia;2. Center for Innovation in Medical Engineering, Faculty of Engineering, University of Malaya, Kuala Lumpur 50603, Malaysia;3. Department of Molecular Medicine, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia;4. Philips Materials Analysis, High Tech Campus 11, 5656 AE Eindhoven, Netherlands;5. Faculty of Health, Medicine and Life Sciences, Maastricht University, Netherlands;1. Institute of Biophysics, Academy of Sciences of the Czech Republic v. v. i., Královopolská 135, 612 65 Brno, Czech Republic;2. Institute of Experimental Biology, Department of Physiology and Immunology of Animals, Faculty of Science, Masaryk University, Kotlářská 2, 611 37 Brno, Czech Republic;3. Department of Histology and Embryology, Faculty of Medicine, Masaryk University, Brno, Czech Republic;4. International Clinical Research Center – Center of Biomolecular and Cellular Engineering, St. Anne''s University Hospital Brno, Brno, Czech Republic;3. Department of Neurology, University of Colorado at Denver, Anschutz Medical Campus, Aurora, Colorado 80045;6. Department of Ophthalmology, University of Colorado at Denver, Anschutz Medical Campus, Aurora, Colorado 80045;4. Department III-Developmental Genetics, Max Planck Institute for Heart and Lung Research, Ludwigstrasse 43, 61231 Bad Nauheim, Germany;5. Departments of Medicine and Physiology, University of California, San Francisco, San Francisco, California 94143;1. Tianjin Key Laboratory for Modern Drug Delivery and High-Efficiency, School of Pharmaceutical Science and Technology, Tianjin University, Tianjin, 300072, China;2. Department of Chemistry, Xinzhou Teachers University, Xinzhou, 034000, China;3. Pharmacy College, Key Laboratory of Hui Ethnic Medicine Modernization, Ministry of Education, Ningxia Medical University, Yinchuan, 750004, China;4. Xi’an University of Technological Information, Xi’an, 710299, China;1. Department of Pharmacology, Shandong University School of Medicine, Jinan, Shandong, 250012 PR China;2. Department of Gastrointestinal Surgery, Provincial Hospital Affiliated to Shandong University, Jinan, Shandong, 250021 PR China;3. Department of Dermatology, The Second Affiliated Hospital of Shandong University, Jinan, Shandong, 250033 PR China;1. College of Biotechnology and Bioresources, Da-Yeh University, 168 University Rd., Dacun, Chang-Hua, Taiwan, Republic of China;2. Department of Medicinal Botanicals and Health Applications, Da-Yeh University, 168 University Rd., Dacun, Chang-Hua, Taiwan, Republic of China;3. Department of Bioresources, Da-Yeh University, 168 University Rd., Dacun, Chang-Hua, Taiwan, Republic of China;4. Department of Bioindustry Technology, Da-Yeh University, 168 University Rd., Dacun, Chang-Hua, Taiwan, Republic of China;5. Department of Food Science and Biotechnology, National Chung Hsing University, 145 Xingda Rd., South Dist., Taichung City, Taiwan, Republic of China
Abstract:A method has been developed for the rapid isolation of platelet membrane glycoproteins (GP) IIb and IIIa. This method produces an excellent yield and does not require the prior isolation of platelet membranes. Outdated platelets were washed and solubilized in Triton X-100. Concanavalin A affinity chromatography was used to purify a platelet glycoprotein fraction. The concanavalin A-retained glycoproteins were eluted and adsorbed with a heparin-Sepharose column to remove a major contaminant, thrombospondin. Sephacryl S-300 gel filtration was used as the final purification step to remove most fibrinogen and low-molecular-weight contaminants. Wheat germ agglutinin affinity chromatography was used to completely remove trace amounts of fibrinogen. The purified GP IIb and GP IIIa were analyzed by sucrose gradient sedimentation and found to consist of heterodimer complexes.
Keywords:
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