Production of tetanus toxin by using media substantially free from meat and blood
DOI:
https://doi.org/10.22159/ajpcr.2016.v9i6.14410Abstract
The present study was to redesign the conventional Mueller and Miller medium to produce tetanus toxin from Clostridium tetani. Meat based ingredients (such as Bovine Heart/ Brain/ Liver Infusion) were replaced with vegetable peptone & alternate casein hydrolysate and scaled up from 100mL to 1000mL. Modified Mueller and Miller Medium containing vegetable peptone (substitute of BHI) and alternate casein hydrolysate were used for production and scale -up of tetanus toxin. Detoxification of tetanus toxin was carried out by using formaldehyde to produce tetanus toxoid. Purification of tetanus toxoid was achieved by fractional precipitation. It was found that under optimum conditions, use of meat free media leads to production of tetanus toxin with equal limes flocculation (Lf) titer and high antigenic content at par with conventional meat based media without any post vaccination infections. The yield of toxin was improved during scale-up of the process. The present study provides a method for growth of Clostridium tetani that maximizes tetanus toxin production without any use of animal-derived components.
Downloads
References
Demain AL, Gerson DF, Fang A. Effective levels of tetanus toxin can be made in a production medium totally lacking both animal (e.g., brain heart infusion) and dairy proteins or digests (e.g., casein hydrolysates). Vaccine 2005; 23:5420-5423.
Demain AL, George S, Kole M, Gerson DF, Fang A. Tetanus toxin production in soy-based medium: nutritional studies and scale-up into small fermentors. Lett Appl Microbiol 2007; 45:635–638.
Fang A, Gerson DF, Demain AL. Menstrum for Culture Preservation and Medium for Seed Preparation in a Tetanus Toxin Production process containing no animal or dairy products. Lett. Appl. Microbiol 2006; 43:360-363.
Fang A, Gerson DF, Demain A L. Production of Clostridium difficile toxin in a medium totally free of both animal and dairy proteins or digests. PNAS 2009; 106 (32): 13225–13229.
Latham WC, Bent DF, Levine L. Tetanus toxin production in the absence of protein. Appl Microbiol 1962; 10:146-152.
Moloney PJ, Hennessy JN. Titration of Tetanal Toxins and Toxoids by Flocculation. Am Assoc Immnol 1944; 48:345—354.
Mueller JH, Miller PA. Variable factors influencing the produc¬tion of tetanus toxin. J Bacteriol 1954; 67:271-277.
Muniandi C, Kavaratty RM. and Subashkumaret R. Large scale recovery of tetanus toxin and toxoid from fermentation broth by microporous tangential flow filtration. International Journal for Biotechnology and Molecular Biology Research 2013; 4(2): 28-37.
Muniandi C, Lakshmanan L, Kavaratty MR, Kumar R. Standardization of process for increased production of pure and potent tetanus toxin. JMID 2013; 3 (3): 133-140.
Ramon. Flocculation dans un me´lange neuter de toxine–antitoxine diphtherique. C R Sceanc Soc Biol 1922;661–663.
Yamamoto A, Akama K. Studies on the side effects of tetanus toxoid. Sensitizing ability of substances present in the medium. Japan J Bacteriol 1969; 24: 359–364.
World Health Organization, 2009. State of the World’s Vaccines and Immunization, Third Ed. http://apps.who.int/iris/bitstream/10665/44169/1/9789241563864_eng.pdf.
World Health Organization, 2012. Recommendations to assure the quality, safety and efficacy of tetanus vaccines (adsorbed), Technical Report Series 980
Published
How to Cite
Issue
Section
The publication is licensed under CC By and is open access. Copyright is with author and allowed to retain publishing rights without restrictions.
