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

Figure 5

Characterization of monoclonal antibodies to rCPB2 by an immunoblotting assay. Monoclonal antibodies (McAbs) produced by hybridoma cell clones listed in Figure 4(b) were tested for their cross-reactions with rCPB2 and native CPB2 in terms of an immunoblotting assay. (a) Images of immunoblots of monoclonal antibodies produced by 2H7, 2G7, and 1E23 hybridoma cell clones against 0.1 μg of rCPB2 with indicated dilutions of ascites generated from Balb/C mice. (b) The sensitivity of immunoblotting assay in detection of rCPB2 using McAb 1E23 at a dilution of 800. (c) The sensitivity of immunoblotting assay in detection of native CPB2 in the culture supernatants and cell lysates of C. perfringens type B (C58-1) strain (left panel) and type C (C59-44) strain (right panel) McAb 1E23 at a dilution of 800. (d) A representative image of ethidium bromide (EB) stained agarose gel of genomic DNA and PCR product of cpb2 gene fragment. Lines 1 and 2 were respective C59-44 genomic DNA and PCR fragment of cpb2 genes and lines 3 and 4 were C58-1 genomic DNA and PCR product, respectively. The immunoblotting results showed that McAb 1E23 could react with both recombinant CPB protein and native CPB2 produced by Clostridium perfringens type C.

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