The Generation and Characterization of Recombinant Protein and Antibodies of Clostridium perfringens Beta2 Toxin

Figure 6

Characterizations of McAbs to CPB2 by an ELISA and abilities to neutralize rCPB2-induced cytotoxicity in vitro. (a) Titration of ascites of McAbs to CPB2 determined by an ELISA. An ELISA plate was coated with rCPB2 at concentration of 3.0 μg/mL; ascites of McAbs were diluted by 2-fold series dilution and incubated with above coated antigens; a reaction of antigen-antibody was detected by an ELISA. The result indicated that McAbs 1E23, 2G7, and 2H7 could be used for detecting CPB by an ELISA. (b) A capacity of McAbs to neutralize the toxicity of rCPB2 toxin in vitro. A 2x LD50 (30 μg/mL) of rCPB2 toxin was mixed with 200 μL of 50x diluted McAbs and incubated at 37°C for 2 h; the mixture was then applied to NCM460 cells incubated at 37°C for additional 12 h. The cell viability was measured by an MTT assay. The result showed abilities of tested McAbs to neutralize the cytotoxicity of rCPB2 in vitro. Among these McAbs, 1E23 displayed the most potential to neutralize the cytotoxicity of rCPB at a dose of 30 μg/mL.

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