FRACTIONATION AND CHARACTERIZATION OF HUMAN OCULAR MUCUS

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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.