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. 2017 Dec 15;11(12):e0006138.
doi: 10.1371/journal.pntd.0006138. eCollection 2017 Dec.

Analysis of the efficacy of Taiwanese freeze-dried neurotoxic antivenom against Naja kaouthia, Naja siamensis and Ophiophagus hannah through proteomics and animal model approaches

Affiliations

Analysis of the efficacy of Taiwanese freeze-dried neurotoxic antivenom against Naja kaouthia, Naja siamensis and Ophiophagus hannah through proteomics and animal model approaches

Chien-Chun Liu et al. PLoS Negl Trop Dis. .

Abstract

In Southeast Asia, envenoming resulting from cobra snakebites is an important public health issue in many regions, and antivenom therapy is the standard treatment for the snakebite. Because these cobras share a close evolutionary history, the amino acid sequences of major venom components in different snakes are very similar. Therefore, either monovalent or polyvalent antivenoms may offer paraspecific protection against envenomation of humans by several different snakes. In Taiwan, a bivalent antivenom-freeze-dried neurotoxic antivenom (FNAV)-against Bungarus multicinctus and Naja atra is available. However, whether this antivenom is also capable of neutralizing the venom of other species of snakes is not known. Here, to expand the clinical application of Taiwanese FNAV, we used an animal model to evaluate the neutralizing ability of FNAV against the venoms of three common snakes in Southeast Asia, including two 'true' cobras Naja kaouthia (Thailand) and Naja siamensis (Thailand), and the king cobra Ophiophagus hannah (Indonesia). We further applied mass spectrometry (MS)-based proteomic techniques to characterize venom proteomes and identify FNAV-recognizable antigens in the venoms of these Asian snakes. Neutralization assays in a mouse model showed that FNAV effectively neutralized the lethality of N. kaouthia and N. siamensis venoms, but not O. hannah venom. MS-based venom protein identification results further revealed that FNAV strongly recognized three-finger toxin and phospholipase A2, the major protein components of N. kaouthia and N. siamensis venoms. The characterization of venom proteomes and identification of FNAV-recognizable venom antigens may help researchers to further develop more effective antivenom designed to block the toxicity of dominant toxic proteins, with the ultimate goal of achieving broadly therapeutic effects against these cobra snakebites.

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Conflict of interest statement

The authors have declared that no competing interests exist.

Figures

Fig 1
Fig 1. RP-HPLC separation of proteins from each cobra venom.
The crude venoms of (A) N. atra, (B) N. kaouthia, (C) N. siamensis, and (D) O. hannah were fractionated on a C18 column; each fraction was collected manually. Shown are the chromatographic patterns for the four HPLC-separated venoms.
Fig 2
Fig 2. Characterization of HPLC-separated, FNAV-recognizable N. atra venom proteins by LC-MS/MS and Western blot analyses.
HPLC-separated fractions of N. atra venom proteins were analyzed by SDS-PAGE, followed by (A) Coomassie Blue staining and (B) Western blotting using FNAV as a probe.
Fig 3
Fig 3. Pie charts showing the relative abundance of cobra venom protein families.
The composition of venom proteins from (A) N. naja, (B) N. kaouthia, (C) N. siamensis, and (D) O. hannah. Results are based on venom protein identification and HPLC peak intensity, and show the major protein components, expressed as a percentage.
Fig 4
Fig 4. Characterization of HPLC-separated, FNAV-recognizable N. kaouthia venom proteins by LC-MS/MS and Western blot analyses.
HPLC-separated fractions of N. kaouthia venom proteins were analyzed by SDS-PAGE, followed by (A) Coomassie Blue staining and (B) Western blotting using FNAV as a probe.
Fig 5
Fig 5. Characterization of HPLC-separated, FNAV-recognizable N. siamensis venom proteins by LC-MS/MS and Western blot analyses.
HPLC-separated fractions of N. siamensis venom proteins were analyzed by SDS-PAGE, followed by (A) Coomassie blue staining and (B) Western blotting using FNAV as a probe.
Fig 6
Fig 6. Characterization of HPLC-separated, FNAV-recognizable O. hannah venom proteins by LC-MS/MS and western blot analyses.
HPLC-separated fractions of O. hannah venom proteins were analyzed by SDS-PAGE, followed by (A) Coomassie Blue staining and (B) Western blotting using FNAV as a probe.

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