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Preparation and Identification of Monoclonal Antibody against Soybean Agglutinin

ZHANG Hai-quan1,2, CHE Dong-sheng1,2, LIU Fei-fei1,2, MU Cheng-long1,2, GU Lin-lin1,2, SUN Ze-wei1, QIN Gui-xin1   

  1. 1. Key Laborotory of Animal Production and Product Quality and Security, Ministry of Education, Jilin Agricultural University, Changchun 130118, China;
    2. College of Animal Science and Technology, Jilin Agricultural University, Changchun 130118, China
  • Received:2012-02-29 Online:2012-09-20 Published:2012-09-18

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Abstract: To prepare hybridoma cells secreting specific monoclonal antibody against soybean agglutinin(SBA),the BALB/c mice were immunized by the purified SBA. The spleen cells isolated from the mice in the highest titer were fused with SP2/0 myeloma cells after three days of last immunization. Two hybridoma cell lines secreting McAb against SBA were established after screening by ELISA and cloning four times by limiting dilution, named as A7,F12. The titers of secreting antibody from cell culture supernatants were both above 1∶2×103,while above 1∶1×106 from ascites of the mice. The subtypes of the monoclonal antibodies were both IgG2b,the molecular weight was 189.6 ku,the affinity constant was 7.1×107 mol/L, Western blotting showed high specificity with SBA. The preparation of the McAb against SBA provided an useful tool for quantitative determination of SBA.

Key words: soybean agglutinin; monoclonal antibody; preparation; identification

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