Ophiophagus hannah venom: proteome, components bound by Naja kaouthia antivenin and neutralization by N. kaouthia neurotoxin-specific human ScFv.
Venomous snakebites are an important health problem in tropical and subtropical countries. King cobra (Ophiophagus hannah) is the largest venomous snake found in South and Southeast Asia. In this study, the O. hannah venom proteome and the venom components cross-reactive to N. kaouthia monospecific antivenin were studied. O. hannah venom consisted of 14 different protein families, including three finger toxins, phospholipases, cysteine-rich secretory proteins, cobra venom factor, muscarinic toxin, L-amino acid oxidase, hypothetical proteins, low cysteine protein, phosphodiesterase, proteases, vespryn toxin, Kunitz, growth factor activators and others (coagulation factor, endonuclease, 5'-nucleotidase). N. kaouthia antivenin recognized several functionally different O. hannah venom proteins and mediated paratherapeutic efficacy by rescuing the O. hannah envenomed mice from lethality. An engineered human ScFv specific to N. kaouthia long neurotoxin (NkLN-HuScFv) cross-neutralized the O. hannah venom and extricated the O. hannah envenomed mice from death in a dose escalation manner. Homology modeling and molecular docking revealed that NkLN-HuScFv interacted with residues in loops 2 and 3 of the neurotoxins of both snake species, which are important for neuronal acetylcholine receptor binding. The data of this study are useful for snakebite treatment when and where the polyspecific antivenin is not available. Because the supply of horse-derived antivenin is limited and the preparation may cause some adverse effects in recipients, a cocktail of recombinant human ScFvs for various toxic venom components shared by different venomous snakes, exemplified by the in vitro produced NkLN-HuScFv in this study, should contribute to a possible future route for an improved alternative to the antivenins.
Graduate Program in Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand. witchu.joy@hotmail.com.
Witchuda
W
Danpaiboon
Department of Molecular Tropical Medicine and Genetics, Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, Thailand. onrapak.rea@mahidol.ac.th.
Onrapak
O
Reamtong
Department of Research and Development, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand. nitat.soo@mahidol.ac.th.
Nitat
N
Sookrung
Laboratory for Research and Technology Development, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand. watee.see@gmail.com.
Watee
W
Seesuay
Laboratory for Research and Technology Development, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand. jub922@yahoo.com.
Yuwaporn
Y
Sakolvaree
Laboratory for Research and Technology Development, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand. gskmu@hotmail.com.
Jeeraphong
J
Thanongsaksrikul
Center for Agriculture Biotechnology and Department of Veterinary Pathology, Faculty of Veterinary Medicine, Kasetsart University, Kam-paeng-saen Campus, Nakhon-pathom 73140, Thailand. fddvt66@outlook.com.
Fonthip
F
Dong-din-on
Graduate Program in Biomedical Science, Faculty of Allied Health Sciences, Thammasat University, Pathumthani 12120, Thailand. srimanote_p@y7mail.com.
Potjanee
P
Srimanote
Laboratory for Research and Technology Development, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand. mam_mt41@hotmail.com.
Kanyarat
K
Thueng-in
Laboratory for Research and Technology Development, Department of Parasitology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand. wanpen.cha@mahidol.ac.th.
Wanpen
W
Chaicumpa
D000818:Animals; D000997:Antivenins / Q000378:metabolism*; D002853:Chromatography, Liquid; D004546:Elapid Venoms / Q000737:chemistry* / Q000378:metabolism / Q000633:toxicity; D017815:Elapidae; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007928:Lethal Dose 50; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D009498:Neurotoxins / Q000276:immunology / Q000494:pharmacology*; D009500:Neutralization Tests; D020543:Proteome; D057127:Single-Chain Antibodies / Q000276:immunology*; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry
D000997:Antivenins; D004546:Elapid Venoms; D009498:Neurotoxins; D020543:Proteome; D057127:Single-Chain Antibodies