Characterization of production processes for tetanus and diphtheria anatoxins.
Tetanus and diphtheria are diseases that still cause significant morbidity and mortality. Clostridium tetani produces the tetanus toxin, a 150-kDa protein. The diphtheria toxin is synthesized by Corynebacterium diphtheriae as a protein of 58 kDa. The objective of this study was to carry out a chemical characterization of the tetanus and diphtheria toxin forms in the several production process stages, and thus to establish an affordable alternative in vitro quality control to aggregate to the classical tests. The 150 kDa band of the tetanus toxin and approximately 58 kDa band of the diphtheria toxin were observed by electrophoresis similar as that described in the literature. The same band of 58 KDa was detected in Western blotting reactions. The results obtained for diphtheria toxin showed very similar protein profiles between distinct lots. For the tetanus toxin, the profiles of the initial stage showed some variability, but the ones of the following stages were similar. The similarity of the electrophoresis results indicated reproduction and consistency of the production processes in Butantan Institute and correlated with the yield and antigenic purity classical data. The establishment of alternative in vitro quality control tests can significantly contribute to achieve the consistency approach supported by WHO.
Divisão de Desenvolvimento Tecnológico e Produção, Instituto Butantan, Av. Dr. Vital Brasil, 1500 São Paulo, Brazil.
Fabiana Belasco
FB
Guilhen
Divisão de Desenvolvimento Tecnológico e Produção, Instituto Butantan, Av. Dr. Vital Brasil, 1500 São Paulo, Brazil.
Aryene Góes
AG
Trezena
Divisão de Desenvolvimento Tecnológico e Produção, Instituto Butantan, Av. Dr. Vital Brasil, 1500 São Paulo, Brazil.
Sally Muller Affonso
SM
Prado
Divisão de Desenvolvimento Tecnológico e Produção, Instituto Butantan, Av. Dr. Vital Brasil, 1500 São Paulo, Brazil.
Hisako Gondo
HG
Higashi
Divisão de Desenvolvimento Tecnológico e Produção, Instituto Butantan, Av. Dr. Vital Brasil, 1500 São Paulo, Brazil; Laboratório de Bacteriologia, Instituto Butantan, Av. Dr. Vital Brasil, 1500 São Paulo, Brazil. Electronic address: mhsonobe@butantan.gov.br.
Martha Harumi
MH
Sonobe
D000942:Antigens, Bacterial / Q000276:immunology; D015153:Blotting, Western; D004167:Diphtheria Toxin / Q000096:biosynthesis*; D004591:Electrophoresis, Polyacrylamide Gel; D011786:Quality Control; D013744:Tetanus Toxin / Q000096:biosynthesis*
D000942:Antigens, Bacterial; D004167:Diphtheria Toxin; D013744:Tetanus Toxin
Diphtheria toxin; Tetanus toxin; Vaccine production