Lethal toxin of Clostridium sordellii (MLD 150 ng/kg) is one of the most potent Clostridial toxins and is responsible
for most of the diseases including sudden death syndrome in cattle, sheep and toxic shock syndrome, necrotizing
faciitis, neonatal omphalitis and gangrene in humans. Lethal toxin (TcsL) is a single chain protein of about 270 kDa.
In the present study, 1.6 kb DNA fragment encoding for the catalytic domain of TcsL was PCR amplified, cloned in
pQE30 UA vector and expressed in E. coli SG 13009. The expression of recombinant lethal toxin protein (rTcsL) was optimized
and it was purified under native conditions using a single step Ni-NTA affinity chromatography. The purified recombinant
protein was used for the production of polyclonal antibodies in mice and rabbit. The raised antibodies reacted
specifically with the purified rTcsL and intact native lethal toxin on Western blot. The biological activity of the recombinant
protein was tested in HeLa cells where it showed the cytotoxicity. Further, the polyclonal antibodies were used for
in-vitro neutralization of purified rTcsL, acid precipitated C. sordellii and C. difficile native toxins in HeLa cells. Mice
and rabbit anti-rTcsL sera effectively neutralized the cytotoxicity of rTcsL and C. sordellii native toxin but it did not neutralize
the cytotoxicity of C. difficile toxin in HeLa cells.
Keywords: Clostridium sordellii, TcsL, cytotoxicity, TSS
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