FRACTIONATION AND CHARACTERIZATION OF HUMAN OCULAR MUCUS

拟议研究的目的是提供信息， 发展一个清晰的了解人类泪膜稳定性在正常和 患病的眼睛，基于各种泪液成分所起的作用。 我们 建议隔离和/或识别责任组件。 特别 注意粘蛋白的分离。 其他重要撕裂 待研究的成分还包括血浆蛋白、脂质和 内切酶 将应用或发展微技术。 方法 分离和纯化将包括选择性溶剂萃取、凝胶 离子交换层析、区带电泳、梯度洗脱 超离心、免疫学方法等。 微量分析 单个组件将为表征提供基础， 识别. 分离物的化学分析将包括碳水化合物 氨基酸分析仪上的氨基酸和一些氨基糖，以及 脂质成分。 某些重要酶的比活性 将被检测。 单个组件的物理特性将包括 粘度、溶解度、分子量、表面活性和模式 结构元素的结合。 我们对这个问题的初步研究： a）大分子组分的分级和部分表征 来自人眼粘液; B）糖蛋白中的O-糖苷键 从人类眼粘液中分离出来，表明我们将站在一个优秀的 有机会实现我们的目标。

The objective of the proposed research is to yield information which will develop a clear understanding of human tear film stability in normal and diseased eyes, based on the roles taken on by various tear components. We propose to isolate and/or identify the responsible components. Special attention will be paid to the isolation of mucin. Other important tear components to be studied will also include plasma proteins, lipids, and enzymes. Microtechniques will be applied or developed. Methods of isolation and purification will include selective solvent extraction, gel and ion exchange chromatography, zone electrophoresis, gradient ultracentrifugation, immunological methods, and others. Microanalyses of individual components will provide a basis for the characterization and identification. Chemical analyses of isolates will include carbohydrates on GLC, amino acids and some amino sugars on the amino acid analyzer, and lipid components by TLC. Specific activities of some enzymes of importance will be assayed. Physical properties of individual components will include viscosity, solubility, molecular weight, surface activities and mode of binding of structural elements. Our preliminary studies on the subject: a) Fractionation and partial characterization of macromolecular components from human ocular mucus; b) O-Glycosidic linkage in the glycoprotein isolates from human ocular mucus, indicate that we will stand an excellent chance to achieve our objectives.