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Human monoclonal ScFv that blocks sodium ion activity of tetrodotoxin

Monrat Chulanetra1, Jeeraphong Thanongsaksrikul2, Kunan Bangphoomi3, Potjanee Srimanote2, Nitat Sookrung4, Hisao Kurazono5, Kiattawee Choowongkomon3 and Wanpen Chaicumpa6*
  • 1 Graduate Program in Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Thailand
  • 2 Graduate Program in Biomedical Sciences, Faculty of Allied Health Sciences, Thammasat University, Thailand
  • 3 Department of Biochemistry, Faculty of Sciences, Kasetsart University, Thailand
  • 4 Department of Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Thailand
  • 5 Department of Applied Veterinary Medicine and Public Health, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Japan
  • 6 Department of Parasitology, Faculty of Medicine Siriraj hospital, Mahidol University, Thailand

Tetrodotoxin (TTX) intoxication is commonly found in Asia-Pacific regions mainly from eating seafood containing the toxin, such as puffer fish, eggs of horseshoe crabs. TTX blocks sodium ion channel of nerve cell membrane causing paresthesia, paralysis, respiratory failure and rapid death, if not treated promptly and properly. Currently, there is no specific anti-dote and the TTX intoxicated patients received only palliative treatment. In this study, human monoclonal single chain antibody fragments (HuScFv) that block the TTX activity for treatment of the TTX intoxication was invented. Phage clones displaying HuScFv that bound specifically to the TTX was selected from a human ScFv phage display library; using TTX as the phage bio-panning antigen. TTX specific HuScFv were produced and purified from the phage transfected E. coli. HuScFv from two E. coli transformants: s16 and s35 could neutralize the TTX toxicity to neuroblastoma cells ex vivo as well as in the TTX intoxicated mice. Computer homology modeling and molecular docking enlightened the molecular mechanisms of the HuScFv in neutralization of the TTX activity. The HuScFv invented in this study have high potential for developing further for treatment of human TTX intoxication.

Acknowledgements

The work was supported by The National Research Council of Thailand (NRCT) of the fiscal year
2009, the Project for Higher Education Research Promotion and National Research University (NRU)
Development, Office of the Higher Education Commission. Monrat Chulanetra is a scholar of Siriraj
Graduate scholarship and the Royal Golden Jubilee Ph.D. program of The Thailand Research Fund
(TRF); Nitat Sookrung and Potjanee Srimanote are TRF grantees; Wanpen Chaicumpa received
DPG5380001 grant of TRF.

References

Chulanetra M, Sookrung N, Srimanote P, Indrawattana N, Thanongsaksrikul J, Sakolvaree Y, Chongsa-Nguan M, Kurazono H, Chaicumpa W. Toxic marine puffer fish in Thailand seas and tetrodotoxin they contained. Toxins (2011) 3:1349-62.
Chulanetra M, Bangphoomi K, Sookrung N, Thanongsaksrikul J, Srimanote P, Sakolvaree Y, Choowongkomon K, Chaicumpa W. Human ScFv that block sodium ion channel activity of tetrodotoxin. Toxicon (2012) 59: 272-82.
Diener M, Christain B, Ahmed MS, Luckas B. Determination of tetrodotoxin and its analogs in the puffer fish Takifugu ablongus from Bangladesh by hydrophilic interaction chromatography and mass spectrometric detection. Anal Bioanal Chem (2007) 389: 1997–2002.
Kodama M, Sato S. "Puffer Fish Toxin,". In: Bureau of Environmental Health. Ministry of Health and Welfare, Japan. The Manual for the Methods of Food Sanitation Tests. Tokyo: Food Hygienic Association (2005). p. 661–66.
Kogure Z, Tamplin ML, Simidu U, Colewell RR. A tissue culture assay for tetrodotoxin, saxitoxin, and related toxin. Toxicon (1988) 26: 191–97.
Kulkeaw K, Sakolvaree Y, Srimanote P, Tongtawe P, Maneewatch S, Sookrung N, Tungtrongchitr A, Tapchaisri P, Kurazono H, Chaicumpa W. Human monoclonal ScFv neutralize lethal Thai cobra, Naja kouathia, neurotoxin. J Proteomics (2009) 72: 270-82.

Keywords: Puffer fish, Tetrodotoxin, Human single chain antibody, Mouse bio-assay, Sodium Channels, phage display

Conference: 15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013.

Presentation Type: Abstract

Topic: Translational immunology and immune intervention

Citation: Chulanetra M, Thanongsaksrikul J, Bangphoomi K, Srimanote P, Sookrung N, Kurazono H, Choowongkomon K and Chaicumpa W (2013). Human monoclonal ScFv that blocks sodium ion activity of tetrodotoxin. Front. Immunol. Conference Abstract: 15th International Congress of Immunology (ICI). doi: 10.3389/conf.fimmu.2013.02.00532

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Received: 04 Jun 2013; Published Online: 22 Aug 2013.

* Correspondence: Prof. Wanpen Chaicumpa, Department of Parasitology, Faculty of Medicine Siriraj hospital, Mahidol University, Bangkok, Thailand, wanpen.cha@mahidol.ac.th