Miniaturized bioactivity assessment coupled to mass spectrometry for guided purification of bioactives from toad and cone snail

F.A.H. Heus, R.A. Otvos, R.L.E.G. Aspers, R. van Elk, J.I. Halff, A.W. Ehlers, S. Duterte, R.J. Lewis, S Wijmenga, A.B. Smit, W.M.A. Niessen, J. Kool

Research output: Contribution to JournalArticleAcademicpeer-review

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. © 2013 by the authors; licensee MDPI, Basel, Switzerland.
Original languageEnglish
Pages (from-to)139-156
JournalBiology
Volume3
DOIs
Publication statusPublished - 2014

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