ISOLATION OF OLIGOSACCHARIDES--SUGAR-BINDING PROTEINS

碳水化合物与许多重要的生理细胞有关。 表面功能包括细胞-细胞识别、生长控制和 毒素和糖蛋白激素的受体。面临的困难 获得足够的数量并确定有限的 大量具有重要生理意义的细胞表面低聚糖是 对所涉及的结构缺乏了解的主要原因 在这些过程中。由于母乳中含有游离低聚糖 在细胞表面糖脂中也发现了结构，针对这些的抗体 低聚糖可用于灵敏的放射免疫分析以检测 复杂生物样品中的特定碳水化合物。 由于化学方法用于结构分析的异常 低聚糖对某些疾病和遗传性疾病的诊断 它们的新陈代谢，需要费力的提纯和昂贵 仪器，它们一般不适用于筛查、诊断 或疾病的分类。诊断用放射免疫测定法 重要的低聚糖可能是一个有价值的替代品。作为……的基础 在此应用中，针对人低聚糖的特异性抗血清将被 应用于人胎粪神经节苷脂的分析 诊断样本和丰富的胎儿糖脂来源。 结合125I标记碳水化合物的细胞表面糖脂 抗体、霍乱毒素、凝集素等生理上重要的物质 在薄层色谱图上可以检测到碳水化合物结合蛋白 通过灵敏的放射自显影技术。低聚糖从体内释放 功能性糖脂可以通过用NaB[~3H]4和 通过放射免疫结合方法从复杂混合物中分离出来。结构性 使用特异性抗寡糖抗体和特异性 外切糖苷酶的消化将是识别次要的基础， 功能性细胞表面碳水化合物结构。这些方法还将 被应用于血型特异性凝集素的表征和 抗体及其相应的糖脂同种抗原。

Carbohydrates have been implicated in many physiologically important cell surface functions including cell-cell recognition, growth control and receptors for toxins and glycoprotein hormones. The difficulties in obtaining sufficient amounts and determining the structure of the limited quantities of physiologically important cell surface oligosaccharides are major reasons for the lack of knowledge about what structures are involved in these processes. Since free oligosaccharides in human milk contain structures also found in cell surface glycolipids, antibodies against these oligosaccharides can be used in sensitive radioimmunoassays to detect specific carbohydrates in complex biological samples. Since chemical methods for structural analyses of abnormal oligosaccharides, diagnostic for certain diseases and genetic disorders of their metabolism, require laborious purification and expensive instrumentation, they are generally not applicable to screening, diagnosis or classification of diseases. Radioimmunoassays for diagnostically important oligosaccharides may be a valuable alternative. As a basis for this application, specific antisera against human oligosaccharides will be applied to the ganglioside analysis of human meconium, a potential diagnostic specimen and a rich source of fetal glycolipids. Cell surface glycolipids that bind 125I-labeled carbohydrate-specific antibodies, cholera toxin, lectins and other physiologically important carbohydrate-binding proteins can be detected on thin layer chromatograms by sensitive autoradiographic techniques. Oligosaccharides released from functional glycolipids can be labeled by reduction with NaB[3H]4 and isolated from complex mixtures by radioimmune-binding methods. Structural analyses using specific anti-oligosaccharide antibodies and specific exoglycosidase digestion will be fundamental to identifying minor, functional cell surface carbohydrate structures. These methods will also be applied to the characterization of blood group specific lectins and antibodies and their corresponding glycolipid isoantigens.