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. and Kool, Jeroen (2014) Miniaturized bioaffinity assessment coupled to mass spectrometry for guided purification of bioactives from toad and cone snail. Biology, 3 1: 139-156. doi:10.3390/biology3010139


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
Title Miniaturized bioaffinity assessment coupled to mass spectrometry for guided purification of bioactives from toad and cone snail
Journal name Biology   Check publisher's open access policy
ISSN 2079-7737
Publication date 2014-02-13
Sub-type Article (original research)
DOI 10.3390/biology3010139
Open Access Status DOI
Volume 3
Issue 1
Start page 139
End page 156
Total pages 18
Place of publication Basel, Switzerland
Publisher MDPI AG
Collection year 2015
Abstract 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.
Keyword Bufo alvarius
Bufo marinus
Conus textile
Lymnaea stagnalis acetylcholine binding protein (Ls-AChBP)
Nano-liquid chromatography-mass spectrometry (nano-LC-MS)
On-line microfluidics
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2015 Collection
Institute for Molecular Bioscience - Publications
 
Versions
Version Filter Type
Citation counts: Scopus Citation Count Cited 1 times in Scopus Article | Citations
Google Scholar Search Google Scholar
Created: Tue, 17 Jun 2014, 01:53:22 EST by System User on behalf of Institute for Molecular Bioscience