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Isolation and Characterization of Polypeptide Core of Human Adenocarcinoma-Associated Mucin
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Abstract:
To study the molecular properties of polypeptide core of human mucin antigens, a polypeptide core of mucin (apomucin) was isolated from human pancreatic cancer cell line SW1990.The tumor mucin was isolated by a combination of molecular sieve chromatography and CsCl density gradient ultracentrifugation. Anhydrous hydrogen flouride was employed to remove carbohydrate units from purified mucin molecules. The purity of mucin and apomucin were identified by SDS-PAGE, the amino acids of apomucin were analized and confirmed by Western Blot using anti-apomucin antibodies.A bunch of monoclonal antibodies were generated against this apomucin. The results showed that this isolated apomucin consisted of many different cDNA sequences peptides, included MUC-l,MUC-2 and MUC-3. The monoclonal antibodies against this apomucin reacted with peptides of molecular weight from 28 to >90 KD.A11 the five monoclonal antibodies against this apomucin reacted with cancer cell lines and cancer tissues,while only one also reacted with normal tissues.
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