| Автор | Midura, Ronald J. |
| Автор | Salustri, Antonietta |
| Автор | Calabro, Anthony |
| Автор | Yanagishita, Masaki |
| Автор | Hascall, Vincent C. |
| Дата выпуска | 1994 |
| dc.description | Recent literature indicates that specific glycosaminoglycan structures are involved in various biological processes, such as anticoagulation, growth factor activation and viral infection. The initial step in the structural analysis of glycosaminoglycans is a definitive compositional analysis of its characteristic disaccharide repeat structures. Current chromatographic or electrophoretic procedures may have limitations in analysing glycosaminoglycan samples that are in low abundance, contain novel structures that need to be further characterized, or are metabolically labelled from radioactive precursors as a result of biosynthetic experiments. This study presents a new methodology for analysing disaccharides and oligosaccharides derived from chondroitin sulphate, dermatan sulphate and hyaluronan that fulfils the above criteria. The procedure involves the separation of reduced forms of these glycoconjugates on a CarboPac PA1 column using alkaline eluants. This study adopted a strategy which uses specific enzymes to release these disaccharides from their glycosaminoglycan forms. A borohydride reduction reaction was modified to be compatible with the buffer conditions commonly used with these enzymes in order to quantitatively reduce the disaccharides to their alditol forms (thereby stabilizing them to alkaline pH). Chromatography conditions were established which separated all known disaccharide alditol structures from chondroitin sulphate, dermatan sulphate and hyaluronan with extremely high resolution in a single run. Integrated pulsed amperometry was compared to UV absorbance measurement at 232 nm as two sensitive methods for detecting these reduced disaccharides; most of them could be routinely detected in the range of 50–500 ng. Data are presented applying this method to quantify hyaluronan in a biological sample which contains ˜5000 cells and only ˜10 ng of hyaluronan. Additional data are presented to demonstrate that this procedure will also separate oligosaccharide alditols derived from hyaluronan. |
| Формат | application.pdf |
| Издатель | Oxford University Press |
| Копирайт | © Oxford University Press |
| Тема | borohydride reduction |
| Тема | glycosaminoglycans |
| Тема | integrated pulsed amperometry |
| Тема | ORIGINAL ARTICLES |
| Название | High-resolution separation of disaccharide and oligosaccharide alditols from chondroitin sulphate, dermatan sulphate and hyaluronan using CarboPac PA1 chromatography |
| Тип | research-article |
| Electronic ISSN | 1460-2423 |
| Print ISSN | 0959-6658 |
| Журнал | Glycobiology |
| Том | 4 |
| Первая страница | 333 |
| Последняя страница | 342 |
| Аффилиация | Midura Ronald J.; Department of Orthopaedic Surgery, University of Iowa 180 Medical Laboratories |
| Аффилиация | Salustri Antonietta; Department of Sanitrá Pubblica e Biologia Cellulare, Faculty of Medicine, Second University of Rome, |
| Аффилиация | Calabro Anthony; Bone Research Branch, National Institute of Dental Research, National Institutes of Health |
| Аффилиация | Yanagishita Masaki; Bone Research Branch, National Institute of Dental Research, National Institutes of Health |
| Аффилиация | Hascall Vincent C.; Bone Research Branch, National Institute of Dental Research, National Institutes of Health |
| Выпуск | 3 |
