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Biotechnology and Applied Biochemistry (2001) 33, (167–172) (Printed in Great Britain)

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Rabies virus production in non-woven polyester fabric (NWPF) packed-bed reactors

Meneme Gümüdereliolu*¹, Esin Aslankaraolu* and S. smet Gürhan†

*Hacettepe University, Chemical Engineering Department, 06532, Beytepe, Ankara, Turkey, and †Foot and Mouth Disease Institute, 06531, Ankara, Turkey

Abbreviations used: NWPF, non-woven polyester fabric; BHK-An₆₆, baby hamster kidney-Ankara 66; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide; CVS, Challenge Virus Standard; ffu, focus-forming units.

¹ To whom correspondence should be addressed (e-mail menemse@hacettepe.edu.tr).

The production of rabies virus from baby hamster kidney-Ankara 66 (BHK-An₆₆, HUKUK 99050302) monolayer cells was examined in a packed-bed reactor containing non-woven polyester fabric (NWPF) discs. At first, growth characteristics of the cells were determined in static culture. The suspension culture studies were realized in a 1 litre spinner basket with NWPF support. BHK-An₆₆ cells (inoculation density, 5×10⁴ cells·ml^-1) were maintained in the reactor loaded with 5 g·l^-1 carrier. The culture medium was Eagle's minimal essential medium supplemented with 10% (v/v) fetal bovine serum at 37 °C. During the culture, the medium was sampled daily to assess glucose and lactate concentrations. At the end of the 7 day culture period the cell density was found to be 2.2×10⁷ cells·ml^-1, and the reactor was inoculated with 5 ml [1.7×10⁶ focus-forming units (ffu)·ml^-1] of the CVS 11 (Challenge Virus Standard) strain of rabies virus. After a 72 h incubation period, the cultures were stained with fluorescein-conjugated anti-rabies globulin and were observed using a fluorescence microscope. Virus titres determined by the Spearman–Kärber method were 2.2×10⁵ ffu·ml^-1. In conclusion, NWPF packed reactors can be considered as a suitable system for the large-scale production of rabies virus.

Received 28 June 2000/16 December 2000; accepted 18 December 2000

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