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《Acta Pharmaceutica Sinica》 2003-01
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Preparation and characterization of monoclonal antibody against marine sulfated polysaccharide drug poly-mannaguluronic acid

CHEN Kun 1, XIN Xian-liang 1, GENG Mei-yu 1, ZHU Jian-chun 2, YANG Ming 2, LI Yong 2 (1. Marine Drug and Food Institute, Ocean University of Qingdao, Qingdao 266003, China; 2. Biologics Institute of Changchun, Changchun 130012, China)  
Aim To prepare and characterize the monoclonal antibody against poly-mannaguluronic acid (911). Methods A hybridoma cell line was obtained by cloning for 3 times those cells that secret antibody against 911 after the fusion of NS-1 myelome cells with spleen cells from Balb/c mice immunized with a 911-bovine serum albumin conjugate, prepared by reductive amination. Hybridoma was inoculated to Balb/c mouse to induce ascites. The antibody was purified by ammonium sulfate and Protein A - Sepharose CL-4B. DD3 was confirmed to be fusion cells after determination of DNA content with flow cytometry. Competitive inhibitory test and Biacore confirmed the cross-reactivity of the antibody with other endogenous polysaccharides or with alginic acid sodium. Igs′ classes and subclasses were identified by Isostrip TM method. The affinity of DD3 was verified by ELISA. Results A hybridoma cell line secreting monoclonal antibody against 911 (marine sulfate polysaccharide) named DD3 was obtained. The DD3 ascites contained specific antibody with titer over 1.0×10 5. There was no cross-reactivity of these antibodies with other endogenous polysaccharides or with alginic acid sodium. The immunoglobulin subclass of DD3 was IgG2a, κ type. The affinity of DD3 was 2.0×10 8 L·mol -1 . Conclusion One hybirdoma line (DD3) secreting monoclonal antibody against 911 was established to provide a potential method for the pharmacokinetic study of 911.
【Fund】: 国家自然科学基金重点项目 (3 0 13 0 2 0 0 ) ;; 国家自然科学基金资助项目 (3 0 0 70 0 694)
【CateGory Index】: R392
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