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Choo Hock Tan
Protein and Interactomics Laboratory, Department of Molecular Medicine, Faculty of Medicine, University of Malaya, Kuala Lumpur 50603, Malaysia

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Journal article
Published: 10 August 2021 in Toxins
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Inadequate effectiveness of Indian antivenoms in treating envenomation caused by the Spectacled Cobra/Indian Cobra (Naja naja) in Sri Lanka has been attributed to geographical variations in the venom composition. This study investigated the de novo venom-gland transcriptomics and venom proteomics of the Sri Lankan N. naja (NN-SL) to elucidate its toxin gene diversity and venom variability. The neutralization efficacy of a commonly used Indian antivenom product in Sri Lanka was examined against the lethality induced by NN-SL venom in mice. The transcriptomic study revealed high expression of 22 toxin genes families in NN-SL, constituting 46.55% of total transcript abundance. Three-finger toxins (3FTX) were the most diversely and abundantly expressed (87.54% of toxin gene expression), consistent with the dominance of 3FTX in the venom proteome (72.19% of total venom proteins). The 3FTX were predominantly S-type cytotoxins/cardiotoxins (CTX) and α-neurotoxins of long-chain or short-chain subtypes (α-NTX). CTX and α-NTX are implicated in local tissue necrosis and fatal neuromuscular paralysis, respectively, in envenomation caused by NN-SL. Intra-species variations in the toxin gene sequences and expression levels were apparent between NN-SL and other geographical specimens of N. naja, suggesting potential antigenic diversity that impacts antivenom effectiveness. This was demonstrated by limited potency (0.74 mg venom/ml antivenom) of the Indian polyvalent antivenom (VPAV) in neutralizing the NN-SL venom. A pan-regional antivenom with improved efficacy to treat N. naja envenomation is needed.

ACS Style

Kin Wong; Kae Tan; Nget Tan; Christeine Gnanathasan; Choo Tan. Elucidating the Venom Diversity in Sri Lankan Spectacled Cobra (Naja naja) through De Novo Venom Gland Transcriptomics, Venom Proteomics and Toxicity Neutralization. Toxins 2021, 13, 558 .

AMA Style

Kin Wong, Kae Tan, Nget Tan, Christeine Gnanathasan, Choo Tan. Elucidating the Venom Diversity in Sri Lankan Spectacled Cobra (Naja naja) through De Novo Venom Gland Transcriptomics, Venom Proteomics and Toxicity Neutralization. Toxins. 2021; 13 (8):558.

Chicago/Turabian Style

Kin Wong; Kae Tan; Nget Tan; Christeine Gnanathasan; Choo Tan. 2021. "Elucidating the Venom Diversity in Sri Lankan Spectacled Cobra (Naja naja) through De Novo Venom Gland Transcriptomics, Venom Proteomics and Toxicity Neutralization." Toxins 13, no. 8: 558.

Journal article
Published: 23 July 2021 in Toxins
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Snakebite envenomation is a serious neglected tropical disease, and its management is often complicated by the diversity of snake venoms. In Asia, pit vipers of the Ovophis species complex are medically important venomous snakes whose venom properties have not been investigated in depth. This study characterized the venom proteomes of Ovophis convictus (West Malaysia), Ovophis tonkinensis (northern Vietnam, southern China), and Ovophis okinavensis (Okinawa, Japan) by applying liquid chromatography-tandem mass spectrometry, which detected a high abundance of snake venom serine proteases (SVSP, constituting 40–60% of total venom proteins), followed by phospholipases A2, snake venom metalloproteinases of mainly P-III class, L-amino acid oxidases, and toxins from other protein families which were less abundant. The venoms exhibited different procoagulant activities in human plasma, with potency decreasing from O. tonkinensis >O. okinavensis >O. convictus. The procoagulant nature of venom confirms that consumptive coagulopathy underlies the pathophysiology of Ovophis pit viper envenomation. The hetero-specific antivenoms Gloydius brevicaudus monovalent antivenom (GbMAV) and Trimeresurus albolabris monovalent antivenom (TaMAV) were immunoreactive toward the venoms, and cross-neutralized their procoagulant activities, albeit at variably limited efficacy. In the absence of species-specific antivenom, these hetero-specific antivenoms may be useful in treating coagulotoxic envenomation caused by the different snakes in their respective regions.

ACS Style

Choo Tan; Praneetha Palasuberniam; Kae Tan. Snake Venom Proteomics, Immunoreactivity and Toxicity Neutralization Studies for the Asiatic Mountain Pit Vipers, Ovophis convictus, Ovophis tonkinensis, and Hime Habu, Ovophis okinavensis. Toxins 2021, 13, 514 .

AMA Style

Choo Tan, Praneetha Palasuberniam, Kae Tan. Snake Venom Proteomics, Immunoreactivity and Toxicity Neutralization Studies for the Asiatic Mountain Pit Vipers, Ovophis convictus, Ovophis tonkinensis, and Hime Habu, Ovophis okinavensis. Toxins. 2021; 13 (8):514.

Chicago/Turabian Style

Choo Tan; Praneetha Palasuberniam; Kae Tan. 2021. "Snake Venom Proteomics, Immunoreactivity and Toxicity Neutralization Studies for the Asiatic Mountain Pit Vipers, Ovophis convictus, Ovophis tonkinensis, and Hime Habu, Ovophis okinavensis." Toxins 13, no. 8: 514.

Journal article
Published: 05 July 2021 in Comparative Biochemistry and Physiology Part D: Genomics and Proteomics
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Envenomation by two medically important Sundaic pit vipers, Trimeresurus wiroti (Malaysia) and Trimeresurus puniceus (Indonesia), causes hemotoxic syndrome with a potentially fatal outcome. Research on the compositions and antigenicity of these pit viper venoms is however lacking, limiting our understanding of the pathophysiology and treatment of envenomation. This study investigated the venom proteomes of both species through a protein decomplexation strategy, applying C18 reverse-phase high-performance liquid chromatography followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and protein identification through nano-electrospray ionization liquid chromatography-tandem mass spectrometry (nano-ESI-LCMS/MS) of trypsin-digested peptides. The venom antigenicity was profiled against the Thai Green Pit Viper Antivenom (GPVAV, a hetero-specific antivenom), using indirect enzyme-linked immunosorbent assay (ELISA). The venom proteomes of T. wiroti and T. puniceus consisted of 10 and 12 toxin families, respectively. The major proteins were of diverse snake venom serine proteases (19–30% of total venom proteins), snake venom metalloproteinases (17–26%), disintegrins (9–16%), phospholipases A2 (8–28%) and C-type lectins (~8%). These were putative snake toxins implicated in hemorrhage and coagulopathy, consistent with clinical hemotoxicity. GPVAV showed strong immunorecognition toward high and medium molecular weight proteins (e.g., SVMP and PLA2) in both venoms, while a lower binding activity was observed toward small proteins such as disintegrins. Conserved antigenicity in the major hemotoxins supported toxicity cross-neutralization by GPVAV and indicated that the immunorecognition of low molecular weight toxins may be optimized for improved binding efficacy. Taken together, the study provides insights into the pathophysiology and antivenom treatment of envenomation caused by T. wiroti and T. puniceus in the region.

ACS Style

Louisa Pernee Lee; Kae Yi Tan; Choo Hock Tan. Snake venom proteomics and antivenomics of two Sundaic lance-headed pit vipers: Trimeresurus wiroti (Malaysia) and Trimeresurus puniceus (Indonesia). Comparative Biochemistry and Physiology Part D: Genomics and Proteomics 2021, 40, 100875 .

AMA Style

Louisa Pernee Lee, Kae Yi Tan, Choo Hock Tan. Snake venom proteomics and antivenomics of two Sundaic lance-headed pit vipers: Trimeresurus wiroti (Malaysia) and Trimeresurus puniceus (Indonesia). Comparative Biochemistry and Physiology Part D: Genomics and Proteomics. 2021; 40 ():100875.

Chicago/Turabian Style

Louisa Pernee Lee; Kae Yi Tan; Choo Hock Tan. 2021. "Snake venom proteomics and antivenomics of two Sundaic lance-headed pit vipers: Trimeresurus wiroti (Malaysia) and Trimeresurus puniceus (Indonesia)." Comparative Biochemistry and Physiology Part D: Genomics and Proteomics 40, no. : 100875.

Journal article
Published: 09 February 2021 in Toxins
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Envenomation resulted from sea snake bite is a highly lethal health hazard in Southeast Asia. Although commonly caused by sea snakes of Hydrophiinae, each species is evolutionarily distinct and thus, unveiling the toxin gene diversity within individual species is important. Applying next-generation sequencing, this study investigated the venom-gland transcriptome of Hydrophis curtus (spine-bellied sea snake) from Penang, West Malaysia. The transcriptome was de novo assembled, followed by gene annotation and sequence analyses. Transcripts with toxin annotation were only 96 in number but highly expressed, constituting 48.18% of total FPKM in the overall transcriptome. Of the 21 toxin families, three-finger toxins (3FTX) were the most abundantly expressed and functionally diverse, followed by phospholipases A2. Lh_FTX001 (short neurotoxin) and Lh_FTX013 (long neurotoxin) were the most dominant 3FTXs expressed, consistent with the pathophysiology of envenomation. Lh_FTX001 and Lh_FTX013 were variable in amino acid compositions and predicted epitopes, while Lh_FTX001 showed high sequence similarity with the short neurotoxin from Hydrophis schistosus, supporting cross-neutralization effect of Sea Snake Antivenom. Other toxins of low gene expression, for example, snake venom metalloproteinases and L-amino acid oxidases not commonly studied in sea snake venom were also identified, enriching the knowledgebase of sea snake toxins for future study.

ACS Style

Choo Tan; Kae Tan. De Novo Venom-Gland Transcriptomics of Spine-Bellied Sea Snake (Hydrophis curtus) from Penang, Malaysia—Next-Generation Sequencing, Functional Annotation and Toxinological Correlation. Toxins 2021, 13, 127 .

AMA Style

Choo Tan, Kae Tan. De Novo Venom-Gland Transcriptomics of Spine-Bellied Sea Snake (Hydrophis curtus) from Penang, Malaysia—Next-Generation Sequencing, Functional Annotation and Toxinological Correlation. Toxins. 2021; 13 (2):127.

Chicago/Turabian Style

Choo Tan; Kae Tan. 2021. "De Novo Venom-Gland Transcriptomics of Spine-Bellied Sea Snake (Hydrophis curtus) from Penang, Malaysia—Next-Generation Sequencing, Functional Annotation and Toxinological Correlation." Toxins 13, no. 2: 127.

Journal article
Published: 14 January 2021 in Toxins
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The Senegalese cobra, Naja senegalensis, is a non-spitting cobra species newly erected from the Naja haje complex. Naja senegalensis causes neurotoxic envenomation in Western Africa but its venom properties remain underexplored. Applying a protein decomplexation proteomic approach, this study unveiled the unique complexity of the venom composition. Three-finger toxins constituted the major component, accounting for 75.91% of total venom proteins. Of these, cardiotoxin/cytotoxin (~53%) and alpha-neurotoxins (~23%) predominated in the venom proteome. Phospholipase A2, however, was not present in the venom, suggesting a unique snake venom phenotype found in this species. The venom, despite the absence of PLA2, is highly lethal with an intravenous LD50 of 0.39 µg/g in mice, consistent with the high abundance of alpha-neurotoxins (predominating long neurotoxins) in the venom. The hetero-specific VINS African Polyvalent Antivenom (VAPAV) was immunoreactive to the venom, implying conserved protein antigenicity in the venoms of N. senegalensis and N. haje. Furthermore, VAPAV was able to cross-neutralize the lethal effect of N. senegalensis venom but the potency was limited (0.59 mg venom completely neutralized per mL antivenom, or ~82 LD50 per ml of antivenom). The efficacy of antivenom should be further improved to optimize the treatment of cobra bite envenomation in Africa.

ACS Style

Kin Wong; Kae Tan; Nget Tan; Choo Tan. A Neurotoxic Snake Venom without Phospholipase A2: Proteomics and Cross-Neutralization of the Venom from Senegalese Cobra, Naja senegalensis (Subgenus: Uraeus). Toxins 2021, 13, 60 .

AMA Style

Kin Wong, Kae Tan, Nget Tan, Choo Tan. A Neurotoxic Snake Venom without Phospholipase A2: Proteomics and Cross-Neutralization of the Venom from Senegalese Cobra, Naja senegalensis (Subgenus: Uraeus). Toxins. 2021; 13 (1):60.

Chicago/Turabian Style

Kin Wong; Kae Tan; Nget Tan; Choo Tan. 2021. "A Neurotoxic Snake Venom without Phospholipase A2: Proteomics and Cross-Neutralization of the Venom from Senegalese Cobra, Naja senegalensis (Subgenus: Uraeus)." Toxins 13, no. 1: 60.

Signal transduction
Published: 28 August 2020 in PLOS Neglected Tropical Diseases
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The aim of this study was to develop an in vitro assay for use in place of in vivo assays of snake venom lethality and antivenom neutralizing potency. A novel in vitro assay has been developed based on the binding of post-synaptically acting α-neurotoxins to nicotinic acetylcholine receptor (nAChR), and the ability of antivenoms to prevent this binding. The assay gave high correlation in previous studies with the in vivo murine lethality tests (Median Lethal Dose, LD50), and the neutralization of lethality assays (Median Effective Dose, ED50) by antisera against Naja kaouthia, Naja naja and Bungarus candidus venoms. Here we show that, for the neurotoxic venoms of 20 elapid snake species from eight genera and four continents, the in vitro median inhibitory concentrations (IC50s) for α-neurotoxin binding to purified nAChR correlated well with the in vivo LD50s of the venoms (R2 = 0.8526, p < 0.001). Furthermore, using this assay, the in vitro ED50s of a horse pan-specific antiserum against these venoms correlated significantly with the corresponding in vivo murine ED50s, with R2 = 0.6896 (p < 0.01). In the case of four elapid venoms devoid or having a very low concentration of α-neurotoxins, no inhibition of nAChR binding was observed. Within the philosophy of 3Rs (Replacement, Reduction and Refinement) in animal testing, the in vitro α-neurotoxin-nAChR binding assay can effectively substitute the mouse lethality test for toxicity and antivenom potency evaluation for neurotoxic venoms in which α-neurotoxins predominate. This will greatly reduce the number of mice used in toxicological research and antivenom production laboratories. The simpler, faster, cheaper and less variable in vitro assay should also expedite the development of pan-specific antivenoms against various medically important snakes in many parts of the world. Snakebite envenomation is an important public health problem recognized by the World Health Organization (WHO) as a neglected tropical disease affecting about 2 million of poor people of the tropical world. The most effective therapy is the timely administration of efficacious antivenoms which are usually produced in horses. The serum/plasma of horse immunized with snake venoms is purified and tested for its efficacies in neutralizing the target venoms. The neutralization is assayed using mice injected with the venom together with the antivenom. This assay requires about 60 mice for each pair of venom and antivenom. The assay is expensive, laborious, giving highly variable results and is objected on ethical and religious grounds. The present study involves the development of an in vitro assay involving the binding of a snake neurotoxin to a soluble receptor protein called nicotinic acetylcholine receptor. It is shown here that this receptor binding assay gave good correlation with the assay using mice. The test tube assay is simpler, faster, cheaper and less variable when compared with the mouse assay and thus could reduce or even replace the use of life animal. Furthermore, it could expedite the development of effective antivenoms against various venomous snakes in many parts of the world.

ACS Style

Kritsada Pruksaphon; Kae Yi Tan; Choo Hock Tan; Pavinee Simsiriwong; José María Gutiérrez; Kavi Ratanabanangkoon. An in vitro α-neurotoxin—nAChR binding assay correlates with lethality and in vivo neutralization of a large number of elapid neurotoxic snake venoms from four continents. PLOS Neglected Tropical Diseases 2020, 14, e0008581 .

AMA Style

Kritsada Pruksaphon, Kae Yi Tan, Choo Hock Tan, Pavinee Simsiriwong, José María Gutiérrez, Kavi Ratanabanangkoon. An in vitro α-neurotoxin—nAChR binding assay correlates with lethality and in vivo neutralization of a large number of elapid neurotoxic snake venoms from four continents. PLOS Neglected Tropical Diseases. 2020; 14 (8):e0008581.

Chicago/Turabian Style

Kritsada Pruksaphon; Kae Yi Tan; Choo Hock Tan; Pavinee Simsiriwong; José María Gutiérrez; Kavi Ratanabanangkoon. 2020. "An in vitro α-neurotoxin—nAChR binding assay correlates with lethality and in vivo neutralization of a large number of elapid neurotoxic snake venoms from four continents." PLOS Neglected Tropical Diseases 14, no. 8: e0008581.

Journal article
Published: 25 April 2020 in International Journal of Biological Macromolecules
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Envenomation by Naja annulifera (snouted cobra), a non-spitting African cobra, can result in local tissue damage and fatal paralysis but a species-specific antivenom treatment is currently lacking. In this study, we investigated the quantitative proteome of N. annulifera venom, incorporating HPLC and LC-MS/MS to elucidate the venom toxicity. The immunoreactivities and in vivo neutralization activities of two hetero-specific antivenom products (Premium Serums Pan Africa polyvalent antivenom, PANAF and VINS African polyvalent antivenom, VAPAV) against the venom were subsequently examined. N. annulifera venom comprises 10 toxin families, with three-finger toxin (3FTx) being the most abundantly expressed (~78%). Within 3FTx, cytotoxin is the most dominant form and made up three-quarter of the venom bulk (~74%), whereas alpha-neurotoxins constitute <4% of the total venom proteins. Phospholipase A2 was undetected in the venom proteome, consistent with the unusual absence of PLA2 from the venoms of cobras in the Uraeus subgenus. In ELISA, PANAF and VAPAV showed comparable immunoreactivity toward the protein antigens of N. annulifera venom. These antivenoms, despite being raised against hetero-specific venoms, were capable of cross-neutralizing the lethal effect of N. annulifera venom in mice, with PANAF being marginally more potent.

ACS Style

Kae Yi Tan; Kin Ying Wong; Nget Hong Tan; Choo Hock Tan. Quantitative proteomics of Naja annulifera (sub-Saharan snouted cobra) venom and neutralization activities of two antivenoms in Africa. International Journal of Biological Macromolecules 2020, 158, 605 -616.

AMA Style

Kae Yi Tan, Kin Ying Wong, Nget Hong Tan, Choo Hock Tan. Quantitative proteomics of Naja annulifera (sub-Saharan snouted cobra) venom and neutralization activities of two antivenoms in Africa. International Journal of Biological Macromolecules. 2020; 158 ():605-616.

Chicago/Turabian Style

Kae Yi Tan; Kin Ying Wong; Nget Hong Tan; Choo Hock Tan. 2020. "Quantitative proteomics of Naja annulifera (sub-Saharan snouted cobra) venom and neutralization activities of two antivenoms in Africa." International Journal of Biological Macromolecules 158, no. : 605-616.

Journal article
Published: 09 April 2020 in Acta Tropica
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The banded krait, Bungarus fasciatus is a medically important venomous snake in Asia. The wide distribution of this species in Southeast Asia and southern China indicates potential geographical variation of the venom which may impact the clinical management of snakebite envenomation. This study investigated the intraspecific venom variation of B. fasciatus from five geographical locales through a venom decomplexing proteomic approach, followed by toxinological and immunological studies. The venom proteomes composed of a total of 9 toxin families, comprising 22 to 31 proteoforms at varying abundances. The predominant proteins were phospholipase A2 (including beta-bungarotoxin), Kunitz-type serine protease inhibitor (KSPI) and three-finger toxins (3FTx), which are toxins that cause neurotoxicity and lethality. The venom lethality varied with geographical origins of the snake, with intravenous median lethal doses (LD50) ranging from 0.45−2.55 µg/g in mice. The Thai Bungarus fasciatus monovalent antivenom (BFMAV) demonstrated a dose-dependent increasing immunological binding activity toward all venoms; however, its in vivo neutralization efficacy varied vastly with normalized potency values ranging from 3 to 28 mg/g, presumably due to the compositional differences of dominant proteins in the different venoms. The findings support that antivenom use should be optimized in different geographical areas. The development of a pan-regional antivenom may be a more sustainable solution for the treatment of snakebite envenomation.

ACS Style

Yi Lin Hia; Kae Yi Tan; Choo Hock Tan. Comparative venom proteomics of banded krait (Bungarus fasciatus) from five geographical locales: Correlation of venom lethality, immunoreactivity and antivenom neutralization. Acta Tropica 2020, 207, 105460 .

AMA Style

Yi Lin Hia, Kae Yi Tan, Choo Hock Tan. Comparative venom proteomics of banded krait (Bungarus fasciatus) from five geographical locales: Correlation of venom lethality, immunoreactivity and antivenom neutralization. Acta Tropica. 2020; 207 ():105460.

Chicago/Turabian Style

Yi Lin Hia; Kae Yi Tan; Choo Hock Tan. 2020. "Comparative venom proteomics of banded krait (Bungarus fasciatus) from five geographical locales: Correlation of venom lethality, immunoreactivity and antivenom neutralization." Acta Tropica 207, no. : 105460.

Research
Published: 01 January 2020 in Journal of Venomous Animals and Toxins including Tropical Diseases
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The Eastern Russell's viper, Daboia siamensis, is a WHO Category 1 medically important venomous snake. It has a wide but disjunct distribution in Southeast Asia. The specific antivenom, D. siamensis Monovalent Antivenom (DsMAV-Thailand) is produced in Thailand but not available in Indonesia, where a heterologous trivalent antivenom, Serum Anti Bisa Ular (SABU), is used instead. This study aimed to investigate the geographical venom variation of D. siamensis from Thailand (Ds-Thailand) and Indonesia (Ds-Indonesia), and the immunorecognition of the venom proteins by antivenoms. The venom proteins were decomplexed with reverse-phase high-performance liquid chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, followed by in-solution tryptic digestion, nano-liquid chromatography-tandem mass spectrometry and protein identification. The efficacies of DsMAV-Thailand and SABU in binding the various venom fractions were assessed using an enzyme-linked immunosorbent assay optimized for immunorecognition profiling. The two most abundant protein families in Ds-Thailand venom are phospholipase A2 (PLA2) and Kunitz-type serine protease inhibitor (KSPI). Those abundant in Ds-Indonesia venom are PLA2 and serine protease. KSPI and vascular endothelial growth factor were detected in Ds-Thailand venom, whereas L-amino acid oxidase and disintegrin were present in Ds-Indonesia venom. Common proteins shared between the two included snaclecs, serine proteases, metalloproteinases, phosphodiesterases, 5'nucleotidases and nerve growth factors at varying abundances. DsMAV-Thailand exhibited strong immunorecognition of the major protein fractions in both venoms, but low immunoreactivity toward the low molecular weight proteins e.g. KSPI and disintegrins. On the other hand, SABU was virtually ineffective in binding all fractionated venom proteins. D. siamensis venoms from Thailand and Indonesia varied geographically in the protein subtypes and abundances. The venoms, nevertheless, shared conserved antigenicity that allowed effective immunorecognition by DsMAV-Thailand but not by SABU, consistent with the neutralization efficacy of the antivenoms. A specific, appropriate antivenom is needed in Indonesia to treat Russell's viper envenomation.

ACS Style

Thava Malar Changra Lingam; Kae Yi Tan; Choo Hock Tan. Proteomics and antivenom immunoprofiling of Russell’s viper (Daboia siamensis) venoms from Thailand and Indonesia. Journal of Venomous Animals and Toxins including Tropical Diseases 2020, 26, e20190048 .

AMA Style

Thava Malar Changra Lingam, Kae Yi Tan, Choo Hock Tan. Proteomics and antivenom immunoprofiling of Russell’s viper (Daboia siamensis) venoms from Thailand and Indonesia. Journal of Venomous Animals and Toxins including Tropical Diseases. 2020; 26 ():e20190048.

Chicago/Turabian Style

Thava Malar Changra Lingam; Kae Yi Tan; Choo Hock Tan. 2020. "Proteomics and antivenom immunoprofiling of Russell’s viper (Daboia siamensis) venoms from Thailand and Indonesia." Journal of Venomous Animals and Toxins including Tropical Diseases 26, no. : e20190048.

Journal article
Published: 17 December 2019 in Acta Tropica
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The wide distribution of king cobra (Ophiophagus hannah), a medically important venomous snake in Asia could be associated with geographical variation in the toxicity and antigenicity of the venom. This study investigated the lethality of king cobra venoms (KCV) from four geographical locales (Malaysia, Thailand, Indonesia, China), and the immunological binding as well as in vivo neutralization activities of three antivenom products (Thai Ophiophagus hannah monovalent antivenom, OHMAV; Indonesian Serum Anti Bisa Ular, SABU; Chinese Naja atra monovalent antivenom, NAMAV) toward the venoms. The Indonesian and Chinese KCV were more lethal (median lethal dose, LD50 ~0.5 μg/g) than those from Malaysia and Thailand (LD50 ~1.0 μg/g). The antivenoms, composed of F(ab)’2, were variably immunoreactive toward the KCV from all locales, with OHMAV exhibited the highest immunological binding activity. In mice, OHMAV neutralized the neurotoxic lethality of Thai KCV most effectively (normalized potency = 118 mg venom neutralized per g antivenom) followed by Malaysian, Indonesian and Chinese KCV. In comparison, the hetero-specific SABU was remarkably less potent by at least 6 to10 folds, whereas NAMAV appeared to be non-effective. The finding supports that a specific king cobra antivenom is needed for the effective treatment of king cobra envenomation in each region.

ACS Style

Kae Yi Tan; Tzu Shan Ng; Aymeric Bourges; Ahmad Khaldun Ismail; Tri Maharani; Sumana Khomvilai; Visith Sitprija; Nget Hong Tan; Choo Hock Tan. Geographical variations in king cobra (Ophiophagus hannah) venom from Thailand, Malaysia, Indonesia and China: On venom lethality, antivenom immunoreactivity and in vivo neutralization. Acta Tropica 2019, 203, 105311 .

AMA Style

Kae Yi Tan, Tzu Shan Ng, Aymeric Bourges, Ahmad Khaldun Ismail, Tri Maharani, Sumana Khomvilai, Visith Sitprija, Nget Hong Tan, Choo Hock Tan. Geographical variations in king cobra (Ophiophagus hannah) venom from Thailand, Malaysia, Indonesia and China: On venom lethality, antivenom immunoreactivity and in vivo neutralization. Acta Tropica. 2019; 203 ():105311.

Chicago/Turabian Style

Kae Yi Tan; Tzu Shan Ng; Aymeric Bourges; Ahmad Khaldun Ismail; Tri Maharani; Sumana Khomvilai; Visith Sitprija; Nget Hong Tan; Choo Hock Tan. 2019. "Geographical variations in king cobra (Ophiophagus hannah) venom from Thailand, Malaysia, Indonesia and China: On venom lethality, antivenom immunoreactivity and in vivo neutralization." Acta Tropica 203, no. : 105311.

Short communication
Published: 27 June 2019 in Toxicon
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Daboia siamensis monovalent antivenom (DSMAV, Thailand) exhibited comparable immunoreactivity toward the venoms of eastern Russell's vipers from Thailand and Indonesia. It also effectively neutralized the procoagulant and lethal effects of both venoms, showing high potency. The Indonesian heterologous trivalent antivenom SABU (Serum Anti Bisa Ular), however, has very weak immunoreactivity and it failed to neutralize the Russell's viper venoms. DSMAV appears to be the appropriate choice of antivenom to treat Russell's viper envenoming.

ACS Style

Thava Malar Changra Lingam; Kae Yi Tan; Choo Hock Tan. Thai Russell's viper monospecific antivenom is immunoreactive and effective in neutralizing the venom of Daboia siamensis from Java, Indonesia. Toxicon 2019, 168, 95 -97.

AMA Style

Thava Malar Changra Lingam, Kae Yi Tan, Choo Hock Tan. Thai Russell's viper monospecific antivenom is immunoreactive and effective in neutralizing the venom of Daboia siamensis from Java, Indonesia. Toxicon. 2019; 168 ():95-97.

Chicago/Turabian Style

Thava Malar Changra Lingam; Kae Yi Tan; Choo Hock Tan. 2019. "Thai Russell's viper monospecific antivenom is immunoreactive and effective in neutralizing the venom of Daboia siamensis from Java, Indonesia." Toxicon 168, no. : 95-97.

Journal article
Published: 12 June 2019 in Journal of Proteomics
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The Philippine cobra, Naja philippinensis, is a WHO Category 1 venomous snake of medical importance responsible for fatal envenomation in the northern Philippines. To elucidate the venom proteome and pathophysiology of envenomation, N. philippinensis venom proteins were decomplexed with reverse-phase high-performance liquid chromatography, and protein fractions were subsequently digested with trypsin, followed by nano-liquid chromatography-tandem mass spectrometry analysis and data mining. Three-finger toxins (3FTX, 66.63% of total venom proteins) and phospholipases A2 (PLA2, 22.88%) constitute the main bulk of venom proteome. Other proteins are present at low abundances (<4% each); these include metalloproteinase, serine protease, cobra venom factor, cysteine-rich secretory protein, vespryn, phosphodiesterase, 5′ nucleotidase and nerve growth factor. In the three-finger toxin family, the alpha-neurotoxins comprise solely short neurotoxins (SNTX, 44.55%), supporting that SNTX is the principal toxin responsible for neuromuscular paralysis and lethality reported in clinical envenomation. Cytotoxins (CTX) are the second most abundant 3FTX proteins in the venom (21.31%). The presence of CTX correlates with the venom cytotoxic effect, which is more prominent in murine cells than in human cells. From the practical standpoint, SNTX-driven neuromuscular paralysis is significant in N. philippinensis envenomation. Antivenom production and treatment should be tailored accordingly to ensure effective neutralization of SNTX. The venom proteome of Naja philippinensis, the Philippine cobra, is unravelled for the first time. Approximately half the protein bulk of the venom is made up of short neurotoxins (44.5% of total venom proteins). As the only alpha-neurotoxins present in the venom, short neurotoxins are the causative toxins of the post-synaptic blockade and fast-onset neuromuscular paralysis in N. philippinensis envenomation. A substantial amount of cytotoxins (21.31%) was also detected in N. philippinensis venom, supporting that the venom is cytotoxic although the effect is much weaker in human cells compared to murine cells. The finding is consistent with the low incidence of local tissue necrosis in N. philippinensis envenomation, although this does not negate the need for monitoring and care of bite wound in the patients.

ACS Style

Choo Hock Tan; Kin Ying Wong; Ho Phin Chong; Nget Hong Tan; Kae Yi Tan. Proteomic insights into short neurotoxin-driven, highly neurotoxic venom of Philippine cobra (Naja philippinensis) and toxicity correlation of cobra envenomation in Asia. Journal of Proteomics 2019, 206, 103418 .

AMA Style

Choo Hock Tan, Kin Ying Wong, Ho Phin Chong, Nget Hong Tan, Kae Yi Tan. Proteomic insights into short neurotoxin-driven, highly neurotoxic venom of Philippine cobra (Naja philippinensis) and toxicity correlation of cobra envenomation in Asia. Journal of Proteomics. 2019; 206 ():103418.

Chicago/Turabian Style

Choo Hock Tan; Kin Ying Wong; Ho Phin Chong; Nget Hong Tan; Kae Yi Tan. 2019. "Proteomic insights into short neurotoxin-driven, highly neurotoxic venom of Philippine cobra (Naja philippinensis) and toxicity correlation of cobra envenomation in Asia." Journal of Proteomics 206, no. : 103418.

Short communication
Published: 22 February 2019 in Toxicon
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Gel filtration chromatography and gel electrophoresis revealed minimal protein degradation in lyophilized antivenoms which were 2-year expired (Hemato Polyvalent, Neuro Polyvalent; Thailand) and 18-year expired (Hemato Bivalent, Neuro Bivalent; Taiwan). All expired antivenoms retained immunological binding activity, and were able to neutralize the hemotoxic or neurotoxic as well as lethal effects of the homologous snake venoms. The findings show that antivenoms under proper storage conditions may remain relatively stable beyond the indicated shelf life. (< 75 words)

ACS Style

Kae Yi Tan; Sin Teng Liew; Qian Yi Tan; Farah Nadhirah Abdul-Rahman; Nor Izzati Azmi; Si Mui Sim; Nget Hong Tan; Sumana Khomvilai; Visith Sitprija; Choo Hock Tan. Evaluating the physicochemical properties and efficacy of recently expired and aged antivenom products from Thailand and Taiwan. Toxicon 2019, 160, 55 -58.

AMA Style

Kae Yi Tan, Sin Teng Liew, Qian Yi Tan, Farah Nadhirah Abdul-Rahman, Nor Izzati Azmi, Si Mui Sim, Nget Hong Tan, Sumana Khomvilai, Visith Sitprija, Choo Hock Tan. Evaluating the physicochemical properties and efficacy of recently expired and aged antivenom products from Thailand and Taiwan. Toxicon. 2019; 160 ():55-58.

Chicago/Turabian Style

Kae Yi Tan; Sin Teng Liew; Qian Yi Tan; Farah Nadhirah Abdul-Rahman; Nor Izzati Azmi; Si Mui Sim; Nget Hong Tan; Sumana Khomvilai; Visith Sitprija; Choo Hock Tan. 2019. "Evaluating the physicochemical properties and efficacy of recently expired and aged antivenom products from Thailand and Taiwan." Toxicon 160, no. : 55-58.

Communication
Published: 14 February 2019 in Toxins
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The protein abundances of phospholipases A2 in cobra venom proteomes appear to vary among cobra species. To determine the unique distribution of snake venom phospholipases A2 (svPLA2) in the cobras, the svPLA2 activities for 15 cobra species were examined with an acidimetric and a colorimetric assay, using egg yolk suspension and 4-nitro-3-octanoyloxy benzoic acid (NOBA) as the substrate. The colorimetric assay showed significant correlation between svPLA2 enzymatic activities with the svPLA2 protein abundances in venoms. High svPLA2 activities were observed in the venoms of Asiatic spitting cobras (Naja sputatrix, Naja sumatrana) and moderate activities in Asiatic non-spitters (Naja naja, Naja atra, Naja kaouthia), African spitters (subgenus Afronaja), and forest cobra (subgenus Boulengerina). African non-spitting cobras of subgenus Uraeus (Naja haje, Naja annulifera, Naja nivea, Naja senegalensis) showed exceptionally low svPLA2 enzymatic activities. The negligible PLA2 activity in Uraeus cobra venoms implies that PLA2 may not be ubiquitous in all snake venoms. The svPLA2 in cobra envenoming varies depending on the cobra species. This may potentially influence the efficacy of cobra antivenom in specific use for venom neutralization.

ACS Style

Choo Hock Tan; Kin Ying Wong; Nget Hong Tan; Tzu Shan Ng; Kae Yi Tan. Distinctive Distribution of Secretory Phospholipases A2 in the Venoms of Afro-Asian Cobras (Subgenus: Naja, Afronaja, Boulengerina and Uraeus). Toxins 2019, 11, 116 .

AMA Style

Choo Hock Tan, Kin Ying Wong, Nget Hong Tan, Tzu Shan Ng, Kae Yi Tan. Distinctive Distribution of Secretory Phospholipases A2 in the Venoms of Afro-Asian Cobras (Subgenus: Naja, Afronaja, Boulengerina and Uraeus). Toxins. 2019; 11 (2):116.

Chicago/Turabian Style

Choo Hock Tan; Kin Ying Wong; Nget Hong Tan; Tzu Shan Ng; Kae Yi Tan. 2019. "Distinctive Distribution of Secretory Phospholipases A2 in the Venoms of Afro-Asian Cobras (Subgenus: Naja, Afronaja, Boulengerina and Uraeus)." Toxins 11, no. 2: 116.

Journal article
Published: 11 February 2019 in Toxins
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The equatorial spitting cobra, Naja sumatrana, is a distinct species of medically important venomous snakes, listed as WHO Category 1 in Southeast Asia. The diversity of its venom genes has not been comprehensively examined, although a few toxin sequences annotated to Naja sputatrix were reported previously through cloning studies. To investigate this species venom genes’ diversity, de novo venom-gland transcriptomics of N. sumatrana from West Malaysia was conducted using next-generation sequencing technology. Genes encoding toxins represented only 60 of the 55,396 transcripts, but were highly expressed, contributing to 79.22% of total gene expression (by total FPKM) in the venom-glands. The toxin transcripts belong to 21 families, and 29 transcripts were further identified as full-length. Three-finger toxins (3FTx) composed of long, short, and non-conventional groups, constituted the majority of toxin transcripts (91.11% of total toxin FPKM), followed by phospholipase A2 (PLA2, 7.42%)—which are putatively pro-inflammatory and cytotoxic. The remaining transcripts in the 19 families were expressed at extremely low levels. Presumably, these toxins were associated with ancillary functions. Our findings unveil the diverse toxin genes unique to N. sumatrana, and provide insights into the pathophysiology of N. sumatrana envenoming.

ACS Style

Ho Phin Chong; Kae Yi Tan; Nget Hong Tan; Choo Hock Tan. Exploring the Diversity and Novelty of Toxin Genes in Naja sumatrana, the Equatorial Spitting Cobra from Malaysia through De Novo Venom-Gland Transcriptomics. Toxins 2019, 11, 104 .

AMA Style

Ho Phin Chong, Kae Yi Tan, Nget Hong Tan, Choo Hock Tan. Exploring the Diversity and Novelty of Toxin Genes in Naja sumatrana, the Equatorial Spitting Cobra from Malaysia through De Novo Venom-Gland Transcriptomics. Toxins. 2019; 11 (2):104.

Chicago/Turabian Style

Ho Phin Chong; Kae Yi Tan; Nget Hong Tan; Choo Hock Tan. 2019. "Exploring the Diversity and Novelty of Toxin Genes in Naja sumatrana, the Equatorial Spitting Cobra from Malaysia through De Novo Venom-Gland Transcriptomics." Toxins 11, no. 2: 104.

Journal article
Published: 06 February 2019 in Toxins
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Trimeresurus nebularis is a montane pit viper that causes bites and envenomation to various communities in the central highland region of Malaysia, in particular Cameron’s Highlands. To unravel the venom composition of this species, the venom proteins were digested by trypsin and subjected to nano-liquid chromatography-tandem mass spectrometry (LC-MS/MS) for proteomic profiling. Snake venom metalloproteinases (SVMP) dominated the venom proteome by 48.42% of total venom proteins, with a characteristic distribution of P-III: P-II classes in a ratio of 2:1, while P-I class was undetected. Snaclecs constituted the second most venomous protein family (19.43%), followed by snake venom serine proteases (SVSP, 14.27%), phospholipases A2 (5.40%), disintegrins (5.26%) and minor proteins including cysteine-rich secretory proteins, L-amino acid oxidases, phosphodiesterases, 5′-nucleotidases. The venomic profile correlates with local (painful progressive edema) and systemic (hemorrhage, coagulopathy, thrombocytopenia) manifestation of T. nebularis envenoming. As specific antivenom is unavailable for T. nebularis, the hetero-specific Thai Green Pit viper Monovalent Antivenom (GPVAV) was examined for immunological cross-reactivity. GPVAV exhibited good immunoreactivity to T. nebularis venom and the antivenom effectively cross-neutralized the hemotoxic and lethal effects of T. nebularis (lethality neutralizing potency = 1.6 mg venom per mL antivenom). The findings supported GPVAV use in treating T. nebularis envenoming.

ACS Style

Choo Hock Tan; Kae Yi Tan; Tzu Shan Ng; Evan S.H. Quah; Ahmad Khaldun Ismail; Sumana Khomvilai; Visith Sitprija; Nget Hong Tan. Venomics of Trimeresurus (Popeia) nebularis, the Cameron Highlands Pit Viper from Malaysia: Insights into Venom Proteome, Toxicity and Neutralization of Antivenom. Toxins 2019, 11, 95 .

AMA Style

Choo Hock Tan, Kae Yi Tan, Tzu Shan Ng, Evan S.H. Quah, Ahmad Khaldun Ismail, Sumana Khomvilai, Visith Sitprija, Nget Hong Tan. Venomics of Trimeresurus (Popeia) nebularis, the Cameron Highlands Pit Viper from Malaysia: Insights into Venom Proteome, Toxicity and Neutralization of Antivenom. Toxins. 2019; 11 (2):95.

Chicago/Turabian Style

Choo Hock Tan; Kae Yi Tan; Tzu Shan Ng; Evan S.H. Quah; Ahmad Khaldun Ismail; Sumana Khomvilai; Visith Sitprija; Nget Hong Tan. 2019. "Venomics of Trimeresurus (Popeia) nebularis, the Cameron Highlands Pit Viper from Malaysia: Insights into Venom Proteome, Toxicity and Neutralization of Antivenom." Toxins 11, no. 2: 95.

Journal article
Published: 23 December 2018 in Toxins
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The venom proteome of Hydrophis curtus (synonym: Lapemis hardwickii) from Penang, Malaysia was investigated with nano-electrospray ionization-liquid chromatography tandem mass spectrometry (ESI-LCMS/MS) of the reverse-phase high-performance liquid chromatography (HPLC) venom fractions. Thirty distinct protein forms were identified as toxins from ten families. The three major protein families were phospholipase A2 (PLA2, 62.0% of total venom proteins), three-finger toxin (3FTX, 26.33%) and cysteine-rich secretory protein (CRiSP, 9.00%). PLA2 comprises diverse homologues (11 forms), predominantly the acidic subtypes (48.26%). 3FTX composed of one short alpha-neurotoxin (SNTX, 22.89%) and four long alpha-neurotoxins (LNTX, 3.44%). Both SNTX and LNTX were lethal in mice (intravenous LD50 = 0.10 and 0.24 μg/g, respectively) but the PLA2 were non-lethal (LD50 >1 μg/g). The more abundant and toxic SNTX appeared to be the main driver of venom lethality (holovenom LD50 = 0.20 μg/g). The heterologous Sea Snake Antivenom (SSAV, Australia) effectively cross-neutralized the venom (normalized potency = 9.35 mg venom neutralized per g antivenom) and the two neurotoxins in vivo, with the LNTX being neutralized more effectively (normalized potency = 3.5 mg toxin/g antivenom) than SNTX (normalized potency = 1.57 mg/g). SSAV immunorecognition was strong toward PLA2 but moderate-to-weak toward the alpha-neurotoxins, indicating that neutralization of the alpha-neurotoxins should be further improved.

ACS Style

Choo Hock Tan; Kae Yi Tan; Tzu Shan Ng; Si Mui Sim; Nget Hong Tan. Venom Proteome of Spine-Bellied Sea Snake (Hydrophis curtus) from Penang, Malaysia: Toxicity Correlation, Immunoprofiling and Cross-Neutralization by Sea Snake Antivenom. Toxins 2018, 11, 3 .

AMA Style

Choo Hock Tan, Kae Yi Tan, Tzu Shan Ng, Si Mui Sim, Nget Hong Tan. Venom Proteome of Spine-Bellied Sea Snake (Hydrophis curtus) from Penang, Malaysia: Toxicity Correlation, Immunoprofiling and Cross-Neutralization by Sea Snake Antivenom. Toxins. 2018; 11 (1):3.

Chicago/Turabian Style

Choo Hock Tan; Kae Yi Tan; Tzu Shan Ng; Si Mui Sim; Nget Hong Tan. 2018. "Venom Proteome of Spine-Bellied Sea Snake (Hydrophis curtus) from Penang, Malaysia: Toxicity Correlation, Immunoprofiling and Cross-Neutralization by Sea Snake Antivenom." Toxins 11, no. 1: 3.

Journal article
Published: 29 October 2018 in Journal of Proteomics
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The proteome of the Pakistani B. sindanus venom was investigated with reverse-phase HPLC and nano-ESI-LCMS/MS analysis. At least 36 distinct proteins belonging to 8 toxin protein families were identified. Three-finger toxin (3FTx), phospholipase A2 (including β-bungarotoxin A-chains) and Kunitz-type serine protease inhibitor (KSPI) were the most abundant, constituting ~95% of total venom proteins. The other toxin proteins of low abundance are snake venom metalloproteinase (SVMP), L-amino acid oxidase (LAAO), acetylcholinesterase (AChE), vespryn and cysteine-rich secretory protein (CRiSP). The venom was highly lethal to mice with LD50 values of 0.04 μg/g (intravenous) and 0.15 μg/g (subcutaneous). The 3FTx proteins are diverse, comprising kappa-neurotoxins, neurotoxin-like protein, non-conventional toxins and muscarinic toxin-like proteins. Kappa-neurotoxins and β-bungarotoxins represent the major toxins that mediate neurotoxicity in B. sindanus envenoming. Alpha-bungarotoxin, commonly present in the Southeast Asian krait venoms, was undetected. The Indian VINS Polyvalent Antivenom (VPAV) was immunoreactive toward the venom, and it moderately cross-neutralized the venom lethality (potency = 0.25 mg/ml). VPAV was able to reverse the neurotoxicity and prevent death in experimentally envenomed mice, but the recovery time was long. The unique toxin composition of B. sindanus venom may be considered in the formulation of a more effective pan-regional, polyspecific antivenom. Bungarus sindanus, an endemic krait species distributed mainly in the Sindh Province of Pakistan is a cause of snake envenomation. Its specific antivenom is, however, lacking. The proteomic study of its venom revealed a substantial presence of κ-bungarotoxins and β-bungarotoxins. The toxin profile corroborates the potent neurotoxicity and lethality of the venom tested in vivo. The heterologous Indian VINS polyvalent antivenom (VPAV) cross-reacted with B. sindanus venom and cross-neutralized the venom neurotoxicity and lethality in mice, albeit the efficacy was moderate. The findings imply that B. sindanus and the phylogenetically related B. caeruleus of India share certain venom epitopes. Research should be advanced to improve the efficacy spectrum of a pan-regional polyspecific antivenom.

ACS Style

Angeline Mei Feng Oh; Choo Hock Tan; Kae Yi Tan; Naeem H. Quraishi; Nget Hong Tan. Venom proteome of Bungarus sindanus (Sind krait) from Pakistan and in vivo cross-neutralization of toxicity using an Indian polyvalent antivenom. Journal of Proteomics 2018, 193, 243 -254.

AMA Style

Angeline Mei Feng Oh, Choo Hock Tan, Kae Yi Tan, Naeem H. Quraishi, Nget Hong Tan. Venom proteome of Bungarus sindanus (Sind krait) from Pakistan and in vivo cross-neutralization of toxicity using an Indian polyvalent antivenom. Journal of Proteomics. 2018; 193 ():243-254.

Chicago/Turabian Style

Angeline Mei Feng Oh; Choo Hock Tan; Kae Yi Tan; Naeem H. Quraishi; Nget Hong Tan. 2018. "Venom proteome of Bungarus sindanus (Sind krait) from Pakistan and in vivo cross-neutralization of toxicity using an Indian polyvalent antivenom." Journal of Proteomics 193, no. : 243-254.

Protocol
Published: 02 October 2018 in Functional Proteomics
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Snake venoms are complex mixtures of proteins and peptides that play vital roles in the survival of venomous snakes. As with their diverse pharmacological activities, snake venoms can be highly variable, hence the importance of understanding the compositional details of different snake venoms. However, profiling venom protein mixtures is challenging, in particular when dealing with the diversity of protein subtypes and their abundances. Here we described an optimized strategy combining a protein decomplexation method with in-solution trypsin digestion and mass spectrometry of snake venom proteins. The approach involves the integrated use of C18 reverse-phase high-performance liquid chromatography (RP-HPLC), sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and nano-electrospray ionization tandem mass spectrometry (nano-ESI-LC-MS/MS).

ACS Style

Choo Hock Tan; Kae Yi Tan; Nget Hong Tan. A Protein Decomplexation Strategy in Snake Venom Proteomics. Functional Proteomics 2018, 1871, 83 -92.

AMA Style

Choo Hock Tan, Kae Yi Tan, Nget Hong Tan. A Protein Decomplexation Strategy in Snake Venom Proteomics. Functional Proteomics. 2018; 1871 ():83-92.

Chicago/Turabian Style

Choo Hock Tan; Kae Yi Tan; Nget Hong Tan. 2018. "A Protein Decomplexation Strategy in Snake Venom Proteomics." Functional Proteomics 1871, no. : 83-92.

Protocol
Published: 02 October 2018 in Advanced Structural Safety Studies
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Reverse-phase high-performance liquid chromatography is commonly employed as a decomplexing strategy in snake venom proteomics. The chromatographic fractions often contain relatively pure toxins that can be assessed functionally for toxicity level through the determination of their median lethal doses (LD50). Further, antivenom efficacy can be evaluated specifically against these venom fractions to understand the limitation of the antivenom as the treatment for snake envenomation. However, methods of toxicity assessment and antivenom evaluation vary across laboratories; hence there is a need to standardize the protocols and parameters, in particular those related to the neutralizing efficacy of antivenom. This chapter outlines the important in vivo techniques and data interpretation that can be applied in the functional study of snake venom proteomes.

ACS Style

Choo Hock Tan; Kae Yi Tan. Functional Application of Snake Venom Proteomics in In Vivo Antivenom Assessment. Advanced Structural Safety Studies 2018, 153 -158.

AMA Style

Choo Hock Tan, Kae Yi Tan. Functional Application of Snake Venom Proteomics in In Vivo Antivenom Assessment. Advanced Structural Safety Studies. 2018; ():153-158.

Chicago/Turabian Style

Choo Hock Tan; Kae Yi Tan. 2018. "Functional Application of Snake Venom Proteomics in In Vivo Antivenom Assessment." Advanced Structural Safety Studies , no. : 153-158.