Beaty N B, Mello R J
J Chromatogr. 1987 Jul 17;418:187-222. doi: 10.1016/0378-4347(87)80009-9.
This review of the mammalian extracellular matrix polysaccharides covered the glycosaminoglycans (GAGs) and their association into proteoglycans. As they necessarily pertain to the chromatographic and electrophoretic separations of these molecules, the structural features of the five principal GAGs were briefly reviewed. Much of the current structural work as well as the separation technology has been concerned with the sulfation state and copolymeric sequences of the individual classes of GAGs. The separation methods discussed included electrophoresis by agarose, acrylamide and cellulose acetate, high-performance liquid chromatography (HPLC), ion-exchange, gel permeation and biospecific affinity methods. Since detection systems are an integral part of chemical separation technology, current thoughts about the best methods to assay GAGs or detect column fractions were discussed. These included polysaccharide-specific detection systems such as Alcian blue dye, 1,9-dimethylmethylene blue, bovine serum albumin-Coomassie blue, as well as non-specific carbohydrate detection systems such as the carbazole or indole hydrochloride methods. Instrumentation used in the detection of chromatography fractions for these molecules was discussed, since the usual ultraviolet detector, standard with HPLC equipment, is often unsatisfactory. The most sensitive specific detection method for GAGs is the use of monoclonal antibodies, which are only now becoming commercially available. The use of these antibodies, combined with HPLC separation, appears to be the best available biochemical technology for studying the extracellular matrix polysaccharides. Finally, the association between proteoglycans, GAGs and mammalian disease processes was reviewed, emphasizing mucopolysaccharidoses and arthritis. The early detection of both of these diseases is desired for effective counselling and treatment. Many of the methods discussed here have been applied, but others are yet to be tried in efforts to further that goal.
这篇关于哺乳动物细胞外基质多糖的综述涵盖了糖胺聚糖(GAGs)及其与蛋白聚糖的结合。由于必然涉及这些分子的色谱和电泳分离,因此简要回顾了五种主要GAGs的结构特征。当前的许多结构研究以及分离技术都关注于各类GAGs的硫酸化状态和共聚序列。所讨论的分离方法包括琼脂糖、丙烯酰胺和醋酸纤维素电泳、高效液相色谱(HPLC)、离子交换、凝胶渗透和生物特异性亲和方法。由于检测系统是化学分离技术的一个组成部分,因此讨论了目前关于测定GAGs或检测柱馏分的最佳方法的思路。这些方法包括多糖特异性检测系统,如阿尔辛蓝染料、1,9 - 二甲基亚甲蓝、牛血清白蛋白 - 考马斯蓝,以及非特异性碳水化合物检测系统,如咔唑或盐酸吲哚法。讨论了用于检测这些分子色谱馏分的仪器,因为通常与HPLC设备配套的紫外检测器往往不尽人意。检测GAGs最灵敏的特异性检测方法是使用单克隆抗体,而这种抗体目前才开始商业化供应。将这些抗体与HPLC分离相结合使用,似乎是研究细胞外基质多糖的最佳现有生化技术。最后,综述了蛋白聚糖、GAGs与哺乳动物疾病过程之间的关联,重点是黏多糖贮积症和关节炎。有效咨询和治疗需要尽早发现这两种疾病。这里讨论的许多方法已经得到应用,但为了实现这一目标,其他一些方法还有待尝试。