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Production and evaluation of a monoclonal antibody to Clostridium perfringens type D epsilon toxin
M I El-Enbaawy1, Y A Abdalla, A Z Hussein
1Microbiology Department, Faculty of Veterinary Medicine, Cairo University Abbasia, Cairo, Egypt.
Abstract:
A monoclonal antibody (mAb) against C. perfringens type D epsilon toxin was produced in hybridoma tissue culture supernatant and in BALB/C mice ascitic fluid. The mAb reacted with single band of the purified epsilon toxin and efficiently neutralized the lethal effect of epsilon toxin in mice. A competitive ELISA (C-ELISA) was developed using this mAb and used to evaluate C. perfringens type D vaccines. Individual serum samples from vaccinated rabbits were tested using C-ELISA and mouse neutralization test (MNT). The mAb based C-ELISA correlated with the conventional MNT for estimation of antitoxin level.
Insights
A new monoclonal antibody effectively neutralizes Clostridium perfringens type D epsilon toxin. This antibody enables a competitive ELISA test that correlates with traditional mouse neutralization tests for vaccine evaluation.
Area of Science:
- Veterinary immunology
- Monoclonal antibody development
- Toxin neutralization
Background:
- Clostridium perfringens type D epsilon toxin is a potent lethal agent.
- Accurate assessment of antitoxin levels is crucial for vaccine efficacy.
- Existing neutralization tests can be resource-intensive.
Purpose of the Study:
- To produce and characterize a monoclonal antibody against C. perfringens type D epsilon toxin.
- To develop a competitive ELISA (C-ELISA) using the monoclonal antibody for antitoxin detection.
- To validate the C-ELISA against a conventional mouse neutralization test (MNT).
Main Methods:
- Monoclonal antibody production in hybridoma and ascitic fluid.
- Western blot analysis to confirm antibody specificity for epsilon toxin.
- Mouse neutralization test (MNT) to assess in vivo efficacy.
- Competitive ELISA (C-ELISA) development and validation using vaccinated rabbit sera.
Main Results:
- A specific monoclonal antibody was successfully generated against epsilon toxin.
- The monoclonal antibody demonstrated efficient neutralization of epsilon toxin's lethal effects in mice.
- The developed C-ELISA showed a strong correlation with the MNT for quantifying antitoxin levels.
Conclusions:
- The developed monoclonal antibody is a valuable tool for neutralizing C. perfringens type D epsilon toxin.
- The mAb-based C-ELISA provides a reliable and potentially more accessible method for evaluating C. perfringens type D vaccines.
- This C-ELISA offers a promising alternative to traditional MNT for routine antitoxin assessment.
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