Journal of Microbiology and Biotechnology

  • 제21권7호
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  • Pages.719-727
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  • 2011
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  • 1017-7825(pISSN)
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  • 1738-8872(eISSN)
한국미생물·생명공학회 (The Korean Society for Microbiology and Biotechnology)
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Effect of Adjuvants on Antibody Titer of Synthetic Recombinant Light Chain of Botulinum Neurotoxin Type B and its Diagnostic Potential for Botulism

  • Jain, Swati (Biotechnology Division, Defence Research and Development Establishment) ;
  • Ponmariappan, S. (Biotechnology Division, Defence Research and Development Establishment) ;
  • Kumar, Om (Biotechnology Division, Defence Research and Development Establishment) ;
  • Singh, Lokendra (Biotechnology Division, Defence Research and Development Establishment)
  • 투고 : 2010.10.21
  • 심사 : 2011.03.30
  • 발행 : 2011.07.28
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초록

Botulism is a neuroparalytic disease caused by Clostridium botulinum, which produces seven (A-G) antigenically diverse neurotoxins (BoNTs). BoNTs are the most poisonous substances known to humans, with a median lethal dose ($LD_{50}$) of approximately 1 ng/kg of body weight. Owing to their extreme potency and lethality, they have the potential to be used as a bioterrorism agent. The mouse bioassay is the gold standard for the detection of botulinum neurotoxins; however, it requires at least 3-4 days for completion. Attempts have been made to develop an ELISA-based detection system, which is potentially an easier and more rapid method of botulinum neurotoxin detection. The present study was designed using a synthetic gene approach. The synthetic gene encoding the catalytic domain of BoNT serotype B from amino acids 1-450 was constructed with PCR overlapping primers (BoNT/B LC), cloned in a pQE30 UA vector, and expressed in an E. coli M15 host system. Recombinant protein production was optimized at 0.5 mM IPTG final concentration, 4 h post induction, resulting in a maximum yield of recombinant proteins. The immunogenic nature of the recombinant BoNT/B LC protein was evaluated by ELISA. Antibodies were raised in BALB/c mice using various adjuvants. A significant rise in antibody titer (p<0.05) was observed in the Alum group, followed by the Titermax Classic group, Freund's adjuvant, and the Titermax Gold group. These developed high-titer antibodies may prove useful for the detection of botulinum neurotoxins in food and clinical samples.

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피인용 문헌

  1. Production of recombinant botulism antigens: A review of expression systems vol.28, pp.None, 2011, https://doi.org/10.1016/j.anaerobe.2014.06.003
  2. Development of immunodetection system for botulinum neurotoxin serotype E vol.147, pp.6, 2011, https://doi.org/10.4103/ijmr.ijmr_1375_16