ALTEX :: Alternatives to Animal Experiments

1998, Supplement

Development and Prevalidation of Alternative Methods for the Potency Testing of Clostridium Perfringens Vaccines

Elvira Ebert, Volker Oeppling, Esther Werner, and Klaus Cussler

Paul-Ehrlich-Institut, D-Langen

SUMMARY

The requirements for the quality control of Clostridium perfringens vaccines for veterinary use are described in the monograph 363 of the European Pharmacopoeia (Ph. Eur.). In the currently used potency test neutralising antibodies against C. perfringens beta- and epsilon toxin are determined in a mouse neutralisation test (MNT). Two ELISA methods were developed for the replacement of the MNT. Both methods use monoclonal antibodies to determine the quantity of specific antibodies against beta toxin (Capture-ELISA) and epsilon toxin (Competitive-ELISA) in vitro. In parallel to the routine batch potency test in mice, the beta- and epsilon antitoxin levels in 523 samples were estimated in the ELISA procedures. A high specificity and a good reproducibility are evident for both test systems. An interlaboratory prevalidation study was carried out to evaluate the relevance and the transferability of the ELISA procedures. It is concluded that both ELISA systems seems to be suitable alternative methods for assessing the potency of beta- and epsilon toxoid in batches of vaccines for veterinary use.

Keywords: C. perfringens, potency testing, alternative methods, ELISA, prevalidation