Preparation and identification of monoclonal antibody against abrin-a |
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| Authors: | Li Xiao-Bing Yang Wei Zhang Yi Zhang Zhi-Gang Kong Tao Li Dong-Na Tang Jia-Jia Liu Lei Liu Guo-Wen Wang Zhe |
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| Institution: | College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi'an Road, Changchun 130062, People's Republic of China. |
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| Abstract: | BALB/c mice were immunized four times with formalin-prepared abrin-a. Using the polyethylene glycol method, immunized splenocytes were isolated and fused with SP2/0 cells. An indirect ELISA was established and used to detect positive clones secreting monoclonal antibodies (mAbs) against abrin-a. After analysis, three hybridoma clones secreting IgG-subtype mAbs were obtained. The antibodies were purified from the hybridoma growth medium using protein A or G affinity chromatography. Western blot analysis was used to analyze the antigenic epitopes on abrin-a recognized by the mAbs. The mAbs were specific for abrin-a, with no detectable cross-reactivity with several homologous toxins and associated agglutinins. Sandwich ELISA was then developed using these mAbs, which had a detection limit for abrin-a of 7.8 ng/mL. |
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