Procedure for isolation of gangliosides in high yield and purity: simultaneous isolation of neutral glycosphingolipids |
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| Authors: | M C Byrne M Sbaschnig-Agler D A Aquino J R Sclafani R W Ledeen |
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| Affiliation: | 1. Department of Tumor Pathology, Faculty of Medical Sciences, University of Fukui, Eiheiji, Japan;2. Department of Urology, Faculty of Medical Sciences, University of Fukui, Eiheiji, Japan;3. Division of Surgical Pathology, University of Fukui Hospital, Eiheiji, Japan;1. Department of Pathology, Boston University School of Medicine, Boston, MA 02118, USA;2. Department of Epidemiology, Boston University School of Public Health, Boston, MA 02118, USA;3. Channing Division of Network Medicine, Brigham and Women''s Hospital, Boston, MA, USA;4. Department of Pathology, Western General Hospital and the University of Edinburgh, Edinburgh, Scotland EH1;5. Department of Pathology, University of Massachusetts Medical School, Worcester, MA 01606, USA;6. Department of Dermatology, University of Florida, Gainesville, FL 32601, USA;7. Mercer University School of Medicine, Macon, GA 31201, USA;8. James A. Haley VA Center (PLMS), Tampa, FL 33612, USA;9. Department of Pathology and Dermatology, Icahn School of Medicine Mount Sinai, New York, NY 11766, USA;10. Department of Anatomic Pathology, Oakland University William Beaumont School of Medicine and Beaumont Health Systems, Royal Oak, MI 48017, USA;11. Dermatopathology Section, VA Integrated Systems Network (VISN1), Department of Pathology and Laboratory Medicine, West Roxbury, MA 02132, USA;1. Faculty of Pharmaceutical Sciences, Tokushima University, Shomachi 1-78, Tokushima 770-8505, Japan;2. Graduate School of Pharmaceutical Sciences, Tokushima University, Shomachi 1-78, Tokushima 770-8505, Japan;3. Institute of Advanced Medical Sciences, Tokushima University, Kuramoto 3-18, Tokushima 770-8503, Japan;1. Center For Functional Onco-Imaging of Department of Radiological Sciences, University of California, Irvine, CA, USA;2. Department of Radiology, E-Da Hospital and I-Shou University, Kaohsiung, Taiwan;3. Graduate Institute of Biomedical Electronics and Bioinformatics, National Taiwan University, Taipei, Taiwan;4. Department of Medical Imaging, Taichung Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation, Taichung, Taiwan;5. Department of Radiology, School of Medicine, Tzu Chi University, Hualien, Taiwan |
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| Abstract: | While several methods for ganglioside extraction and isolation have been described, relatively little attention has been given to the effectiveness of separation from peptides, phospholipids, and various low-molecular-weight contaminants. A procedure is described for isolation of gangliosides in high purity and good yield from 1- to 400-mg samples (wet wt). A key step was mild acidification following homogenization, designed to dissociate gangliosides from lipophilic peptides which coextracted into organic solvents. This has proved particularly helpful for myelin and myelin-containing tissues (e.g., white matter, nerve) whose proteins have presented special problems in ganglioside isolation. In this study isolation was effected by consecutive chromatographies on Sephadex LH-20, DEAE-Sephadex, and silica gel following the initial acidification. The method applied to bovine white matter gave tissue concentrations (calculated from yields and radiolabeled tracer recoveries) that were similar to those obtained with three previously described procedures; however, peptide contaminants were an order of magnitude lower. Removal of low-molecular-weight contaminants, including nucleotide sugars, was virtually complete. In addition to ganglioside isolation the method can be used to obtain neutral glycosphingolipids as well. It is believed to have broad applicability to a diversity of tissues. |
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