Parallel in vitro ion channel and in vivo zebrafish assaying of elapid snake venoms following chromatographic separation of toxin components

Snake venoms are complex bioactive mixtures designed to paralyse, kill, or digest prey. These venoms are of pharmacological interest due to their ability to modulate molecular targets such as ion channels and receptors with high specificity and potency. Traditional studies often focus on in vitro mo...

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Main Authors: Arif Arrahman, Haifeng Xu, Muzaffar A. Khan, Tijmen S. Bos, Julien Slagboom, Guus C. van der Velden, Ulrike Nehrdich, Nicholas R. Casewell, Michael K. Richardson, Christian Tudorache, Fernanda C. Cardoso, Jeroen Kool
Format: Article
Language:English
Published: Elsevier 2025-07-01
Series:SLAS Discovery
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Online Access:http://www.sciencedirect.com/science/article/pii/S2472555225000322
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author Arif Arrahman
Haifeng Xu
Muzaffar A. Khan
Tijmen S. Bos
Julien Slagboom
Guus C. van der Velden
Ulrike Nehrdich
Nicholas R. Casewell
Michael K. Richardson
Christian Tudorache
Fernanda C. Cardoso
Jeroen Kool
author_facet Arif Arrahman
Haifeng Xu
Muzaffar A. Khan
Tijmen S. Bos
Julien Slagboom
Guus C. van der Velden
Ulrike Nehrdich
Nicholas R. Casewell
Michael K. Richardson
Christian Tudorache
Fernanda C. Cardoso
Jeroen Kool
author_sort Arif Arrahman
collection DOAJ
description Snake venoms are complex bioactive mixtures designed to paralyse, kill, or digest prey. These venoms are of pharmacological interest due to their ability to modulate molecular targets such as ion channels and receptors with high specificity and potency. Traditional studies often focus on in vitro molecular analysis or in vivo behavioural effects, limiting comprehensive understanding. Here, we present a high-throughput screening platform that combines in vitro ion channel assays with in vivo zebrafish larval bioassays using nanofractionation analytics. This method integrates post-column calcium flux assays, zebrafish paralytic bioassays, toxin mass spectrometry, and proteomics to link bioactivity with toxin identification. Using elapid snake venoms (genus Dendroaspis, Naja, and Hemachatus) as a proof of concept, we identified several toxins modulating ion channels with paralytic effects on zebrafish larvae. Our approach enables parallel acquisition of in vitro and in vivo data, offering a robust guide for identifying and characterising ion channel modulators with defined molecular targets.
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spelling doaj-art-92f12c63cbf041c19ddd8a0f4cf56f0b2025-08-20T02:05:10ZengElsevierSLAS Discovery2472-55522025-07-013410023910.1016/j.slasd.2025.100239Parallel in vitro ion channel and in vivo zebrafish assaying of elapid snake venoms following chromatographic separation of toxin componentsArif Arrahman0Haifeng Xu1Muzaffar A. Khan2Tijmen S. Bos3Julien Slagboom4Guus C. van der Velden5Ulrike Nehrdich6Nicholas R. Casewell7Michael K. Richardson8Christian Tudorache9Fernanda C. Cardoso10Jeroen Kool11Amsterdam Institute of Molecular and Life Sciences (AIMMS), Division of Bioanalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, Faculty of Sciences, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands; Centre for Analytical Sciences Amsterdam (CASA), The Netherlands; Faculty of Pharmacy Universitas Indonesia, Depok, Jawa Barat, IndonesiaAmsterdam Institute of Molecular and Life Sciences (AIMMS), Division of Bioanalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, Faculty of Sciences, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands; Centre for Analytical Sciences Amsterdam (CASA), The NetherlandsInstitute Biology Leiden, Leiden University, Leiden, The Netherlands; Department of Microbiology and Molecular Genetics, Bahauddin Zakariya University, Multan, PakistanAmsterdam Institute of Molecular and Life Sciences (AIMMS), Division of Bioanalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, Faculty of Sciences, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands; Centre for Analytical Sciences Amsterdam (CASA), The NetherlandsAmsterdam Institute of Molecular and Life Sciences (AIMMS), Division of Bioanalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, Faculty of Sciences, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands; Centre for Analytical Sciences Amsterdam (CASA), The NetherlandsInstitute Biology Leiden, Leiden University, Leiden, The NetherlandsInstitute Biology Leiden, Leiden University, Leiden, The NetherlandsCentre for Snakebite Research and Interventions, Department of Tropical Disease Biology, Liverpool School of Tropical Medicine, Liverpool, United KingdomInstitute Biology Leiden, Leiden University, Leiden, The NetherlandsInstitute Biology Leiden, Leiden University, Leiden, The NetherlandsInstitute for Molecular Bioscience, The University of Queensland, Brisbane, AustraliaAmsterdam Institute of Molecular and Life Sciences (AIMMS), Division of Bioanalytical Chemistry, Department of Chemistry and Pharmaceutical Sciences, Faculty of Sciences, Vrije Universiteit Amsterdam, Amsterdam, The Netherlands; Centre for Analytical Sciences Amsterdam (CASA), The Netherlands; Corresponding author.Snake venoms are complex bioactive mixtures designed to paralyse, kill, or digest prey. These venoms are of pharmacological interest due to their ability to modulate molecular targets such as ion channels and receptors with high specificity and potency. Traditional studies often focus on in vitro molecular analysis or in vivo behavioural effects, limiting comprehensive understanding. Here, we present a high-throughput screening platform that combines in vitro ion channel assays with in vivo zebrafish larval bioassays using nanofractionation analytics. This method integrates post-column calcium flux assays, zebrafish paralytic bioassays, toxin mass spectrometry, and proteomics to link bioactivity with toxin identification. Using elapid snake venoms (genus Dendroaspis, Naja, and Hemachatus) as a proof of concept, we identified several toxins modulating ion channels with paralytic effects on zebrafish larvae. Our approach enables parallel acquisition of in vitro and in vivo data, offering a robust guide for identifying and characterising ion channel modulators with defined molecular targets.http://www.sciencedirect.com/science/article/pii/S2472555225000322Elapid snake venomzebrafish in vivo assayion channel in vitro assaysmass spectrometrynanofractionationtoxicity assay
spellingShingle Arif Arrahman
Haifeng Xu
Muzaffar A. Khan
Tijmen S. Bos
Julien Slagboom
Guus C. van der Velden
Ulrike Nehrdich
Nicholas R. Casewell
Michael K. Richardson
Christian Tudorache
Fernanda C. Cardoso
Jeroen Kool
Parallel in vitro ion channel and in vivo zebrafish assaying of elapid snake venoms following chromatographic separation of toxin components
SLAS Discovery
Elapid snake venom
zebrafish in vivo assay
ion channel in vitro assays
mass spectrometry
nanofractionation
toxicity assay
title Parallel in vitro ion channel and in vivo zebrafish assaying of elapid snake venoms following chromatographic separation of toxin components
title_full Parallel in vitro ion channel and in vivo zebrafish assaying of elapid snake venoms following chromatographic separation of toxin components
title_fullStr Parallel in vitro ion channel and in vivo zebrafish assaying of elapid snake venoms following chromatographic separation of toxin components
title_full_unstemmed Parallel in vitro ion channel and in vivo zebrafish assaying of elapid snake venoms following chromatographic separation of toxin components
title_short Parallel in vitro ion channel and in vivo zebrafish assaying of elapid snake venoms following chromatographic separation of toxin components
title_sort parallel in vitro ion channel and in vivo zebrafish assaying of elapid snake venoms following chromatographic separation of toxin components
topic Elapid snake venom
zebrafish in vivo assay
ion channel in vitro assays
mass spectrometry
nanofractionation
toxicity assay
url http://www.sciencedirect.com/science/article/pii/S2472555225000322
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