FRACTIONATION AND CHARACTERIZATION OF OCULAR MUCUS

拟议研究的目的是获取信息 使我们更清楚地了解各种眼球的作用 粘液凝胶形成和泪膜中的粘液和泪液成分 健康和疾病的稳定。这将涉及到隔离和 沙门氏菌粘液中负责成分的表征 无论是正常眼睛还是患病眼睛。我们将特别注意 眼部粘蛋白的分离。其他粘液/撕裂成分 需要研究的包括血清和泪液特异性蛋白，脂质， 糖胺多聚糖和蛋白多糖。用于提纯 眼部粘液，标本将根据血型进行分离 个人眼粘液捐赠者的名单。隔离和分离的方法 纯化将包括选择性溶剂提取、亲和力和 离子交换层析、梯度超速离心法和 免疫学方法。化学成分分析 分离株将包括GLC测定的碳水化合物、氨基酸和 氨基酸分析测定氨基酸组成，薄层扫描法测定脂肪成分 和GLC。粘蛋白的结构分析将涉及氨基酸 多肽核心的测序以及GLC、MS和核磁共振 光谱学结合糖分析和免疫学 低聚糖链的分析。抗(抗)单抗 粘蛋白、撕裂前白蛋白，可能还有脱糖粘蛋白 分离株将会准备好。个体的物理性质研究 粘液成分，特别是粘蛋白，将包括 粘度、溶解度、分子的测量/测定 重量、表面活性和结构之间的结合方式 元素。微量分析的最新进展和我们的经验 这些技术的应用将提供一个极好的 为实现这些目标提供了机会。

The object of the proposed research is to acquire information leading to a clearer understanding of the roles of various ocular mucus and tear components in mucus gel formation and tear film stability in health and disease. This will involve isolation and characterization of the responsible components in mucus from both normal and diseased eyes. Special attention will be paid to the isolation of ocular mucin. Other mucus/tear components to be studied include serum and tear specific proteins, lipids, glycosaminoglycans, and proteoglycans. For the purification of ocular mucin, specimens will be separated according to blood type of individual ocular mucus donors. Methods of isolation and purification will include selective solvent extraction, affinity and ion exchange chromatographies, gradient ultracentrifugation, and immunological methods. Chemical compositional analyses of isolates will include carbohydrates by GLC, amino acids and amino sugars by amino acid analysis, and lipid components by TLC and GLC. Structural analyses of mucin will involve amino acid sequencing of the peptide core, and GLC, MS, and NMR spectroscopy in conjunction with sugar analysis and immunological assays of the oligosaccharide chains. (Monoclonal) antibodies to mucin, tear prealbumin, and possibly de-glycosylated mucin isolates will be prepared. Physical property studies of individual mucus components, particularly mucin, will include measurements/determinations of viscosity, solubility, molecular weight, surface activity, and mode of binding among structural elements. Recent advances in microanalyses and our experience with the application of these techniques will provide an excellent opportunity for achieving these objectives.