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Miniaturized Bioaffinity Assessment Coupled to Mass Spectrometry for Guided Purification of Bioactives from Toad and Cone Snail

A nano-flow high-resolution screening platform, featuring a parallel chip-based microfluidic bioassay and mass spectrometry coupled to nano-liquid chromatography, was applied to screen animal venoms for nicotinic acetylcholine receptor like (nAChR) affinity by using the acetylcholine binding protein...

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Autores principales: Heus, Ferry, Otvos, Reka A., Aspers, Ruud L. E. G., van Elk, Rene, Halff, Jenny I., Ehlers, Andreas W., Dutertre, Sébastien, Lewis, Richard J., Wijmenga, Sybren, Smit, August B., Niessen, Wilfried M. A., Kool, Jeroen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4009767/
https://www.ncbi.nlm.nih.gov/pubmed/24833338
http://dx.doi.org/10.3390/biology3010139
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author Heus, Ferry
Otvos, Reka A.
Aspers, Ruud L. E. G.
van Elk, Rene
Halff, Jenny I.
Ehlers, Andreas W.
Dutertre, Sébastien
Lewis, Richard J.
Wijmenga, Sybren
Smit, August B.
Niessen, Wilfried M. A.
Kool, Jeroen
author_facet Heus, Ferry
Otvos, Reka A.
Aspers, Ruud L. E. G.
van Elk, Rene
Halff, Jenny I.
Ehlers, Andreas W.
Dutertre, Sébastien
Lewis, Richard J.
Wijmenga, Sybren
Smit, August B.
Niessen, Wilfried M. A.
Kool, Jeroen
author_sort Heus, Ferry
collection PubMed
description A nano-flow high-resolution screening platform, featuring a parallel chip-based microfluidic bioassay and mass spectrometry coupled to nano-liquid chromatography, was applied to screen animal venoms for nicotinic acetylcholine receptor like (nAChR) affinity by using the acetylcholine binding protein, a mimic of the nAChR. The potential of this microfluidic platform is demonstrated by profiling the Conus textile venom proteome, consisting of over 1,000 peptides. Within one analysis (<90 min, 500 ng venom injected), ligands are detected and identified. To show applicability for non-peptides, small molecular ligands such as steroidal ligands were identified in skin secretions from two toad species (Bufo alvarius and Bufo marinus). Bioactives from the toad samples were subsequently isolated by MS-guided fractionation. The fractions analyzed by NMR and a radioligand binding assay with α7-nAChR confirmed the identity and bioactivity of several new ligands.
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spelling pubmed-40097672014-05-07 Miniaturized Bioaffinity Assessment Coupled to Mass Spectrometry for Guided Purification of Bioactives from Toad and Cone Snail Heus, Ferry Otvos, Reka A. Aspers, Ruud L. E. G. van Elk, Rene Halff, Jenny I. Ehlers, Andreas W. Dutertre, Sébastien Lewis, Richard J. Wijmenga, Sybren Smit, August B. Niessen, Wilfried M. A. Kool, Jeroen Biology (Basel) Article A nano-flow high-resolution screening platform, featuring a parallel chip-based microfluidic bioassay and mass spectrometry coupled to nano-liquid chromatography, was applied to screen animal venoms for nicotinic acetylcholine receptor like (nAChR) affinity by using the acetylcholine binding protein, a mimic of the nAChR. The potential of this microfluidic platform is demonstrated by profiling the Conus textile venom proteome, consisting of over 1,000 peptides. Within one analysis (<90 min, 500 ng venom injected), ligands are detected and identified. To show applicability for non-peptides, small molecular ligands such as steroidal ligands were identified in skin secretions from two toad species (Bufo alvarius and Bufo marinus). Bioactives from the toad samples were subsequently isolated by MS-guided fractionation. The fractions analyzed by NMR and a radioligand binding assay with α7-nAChR confirmed the identity and bioactivity of several new ligands. MDPI 2014-02-13 /pmc/articles/PMC4009767/ /pubmed/24833338 http://dx.doi.org/10.3390/biology3010139 Text en © 2014 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).
spellingShingle Article
Heus, Ferry
Otvos, Reka A.
Aspers, Ruud L. E. G.
van Elk, Rene
Halff, Jenny I.
Ehlers, Andreas W.
Dutertre, Sébastien
Lewis, Richard J.
Wijmenga, Sybren
Smit, August B.
Niessen, Wilfried M. A.
Kool, Jeroen
Miniaturized Bioaffinity Assessment Coupled to Mass Spectrometry for Guided Purification of Bioactives from Toad and Cone Snail
title Miniaturized Bioaffinity Assessment Coupled to Mass Spectrometry for Guided Purification of Bioactives from Toad and Cone Snail
title_full Miniaturized Bioaffinity Assessment Coupled to Mass Spectrometry for Guided Purification of Bioactives from Toad and Cone Snail
title_fullStr Miniaturized Bioaffinity Assessment Coupled to Mass Spectrometry for Guided Purification of Bioactives from Toad and Cone Snail
title_full_unstemmed Miniaturized Bioaffinity Assessment Coupled to Mass Spectrometry for Guided Purification of Bioactives from Toad and Cone Snail
title_short Miniaturized Bioaffinity Assessment Coupled to Mass Spectrometry for Guided Purification of Bioactives from Toad and Cone Snail
title_sort miniaturized bioaffinity assessment coupled to mass spectrometry for guided purification of bioactives from toad and cone snail
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4009767/
https://www.ncbi.nlm.nih.gov/pubmed/24833338
http://dx.doi.org/10.3390/biology3010139
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