International Journal of
Biotechnology and Molecular Biology Research

  • Abbreviation: Int. J. Biotechnol. Mol. Biol. Res.
  • Language: English
  • ISSN: 2141-2154
  • DOI: 10.5897/IJBMBR
  • Start Year: 2010
  • Published Articles: 99

Full Length Research Paper

Large scale recovery of tetanus toxin and toxoid from fermentation broth by microporous tangential flow filtration

Chellamani Muniandi1*, Kavaratty Raju Mani1 and Rathinasamy Subashkumar2
1Pasteur Institute of India, Coonoor-643 103, The Nilgiris, Tamil Nadu, India. 2PG and Research Department of Biotechnology, Kongunadu Arts and Science College, Coimbatore – 641 029, Tamil Nadu, India
Email: [email protected], [email protected]

  •  Accepted: 08 February 2013
  •  Published: 31 May 2013

Abstract

The commercial production of purified tetanus toxoid mainly depends on the effective separation of the bacterial toxin and toxoid from large volumes of fermentation broth of Clostridium tetani (Harvard 49205) vaccine strain. Tangential flow or cross-flow filtration system was used as rapid drive in the processing of immunobiological assays of tetanus toxin. Tetanus toxoid was prepared by detoxifying the culture filtrates of Ctetani and further purified by ultrafiltration, salt fractionation and adsorption onto aluminium phosphate. Present study deals with the separation of tetanus toxins using a microporous membrane (0.22 µm) and concentration of tetanus toxoids using an ultrafiltration membrane (30 kDa, NMWL pore size) with operational variables like average trans-membrane pressure (ATP), cross flow rate, flux. Under the best conditions, >96% recovery was achieved. Additionally, potency control of 10 batches of tetanus toxoid, prepared from the filtered toxins/toxoid lots by microporous tangential flow filtration system, was evaluated by in vitro passive haemagglutination (PHA) assay and the results obtained in the in vitro PHA were compared with in vivo toxin neutralization (TN) test. An excellent correlation between in vitro test and in vivo TN test was observed by Spearman’s correlation coefficient. It reveals that the process development in which employing available equipment and the in vitro PHA is a promising alternative to the toxic TN test in the potency assay of tetanus vaccine.

Key words: Tetanus toxin, Seitz filtration, tangential flow filtration, microfiltration module, flux, tetanus toxoid, immunogenicity test.