3 years ago

Multichannel Open Tubular Enzyme Reactor Online Coupled with Mass Spectrometry for Detecting Ricin

Multichannel Open Tubular Enzyme Reactor Online Coupled with Mass Spectrometry for Detecting Ricin
Steven Ray Wilson, Elsa Lundanes, Siri Enger, Bent-Tore Røen, Ole Kristian Brandtzaeg
For counterterrorism purposes, a selective nano liquid chromatography–mass spectrometry (nanoLC-MS) platform was developed for detecting the highly lethal protein ricin from castor bean extract. Manual sample preparation steps were omitted by implementing a trypsin/Lys-C enzyme-immobilized multichannel reactor (MCR) consisting of 126 channels (8 μm inner diameter in all channels) that performed online digestion of proteins (5 min reaction time, instead of 4–16 h in previous in-solution methods). Reduction and alkylation steps were not required. The MCR allowed identification of ricin by signature peptides in all targeted mode injections performed, with a complete absence of carry-over in blank injections. The MCRs (interior volume ≈ 1 μL) have very low backpressure, allowing for trivial online coupling with commercial nanoLC-MS systems. The open tubular nature of the MCRs allowed for repeatable within/between-reactor preparation and performance.

Publisher URL: http://dx.doi.org/10.1021/acs.analchem.7b02590

DOI: 10.1021/acs.analchem.7b02590

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