Norharmane matrix enhances detection of endotoxin by MALDI-MS for simultaneous profiling of pathogen, host, and vector systems

Scott, A.J., Flinders, B., Cappell, J., Liang, T., Pelc, R.S., Tran, B., Kilgour, D.P.A. ORCID: 0000-0002-3860-7532, Heeren, R.M.A., Goodlett, D.R. and Ernst, R.K., 2016. Norharmane matrix enhances detection of endotoxin by MALDI-MS for simultaneous profiling of pathogen, host, and vector systems. Pathogens and Disease, ftw097. ISSN 2049-632X

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Abstract

The discovery of novel pathogenic mechanisms engaged during bacterial infections requires the evolution of advanced techniques. Here, we evaluate the dual polarity matrix norharmane (NRM) to improve detection of bacterial lipid A (endotoxin), from host and vector tissues infected with Francisella novicida (Fn). We evaluated NRM for improved detection and characterization of a wide range of lipids in both positive and negative polarities, including lipid A and phospholipids across a range of matrix assisted laser desorption-ionization (MALDI)-coupled applications. NRM matrix improved the limit of detection (LOD) for monophosphoryl lipid A (MPLA) down to picogram-level representing a ten-fold improvement of LOD versus 2,5-dihydroxybenzoic acid (DHB) and 100-fold improvement of LOD versus 9-aminoacridine (9-AA). Improved LOD for lipid A subsequently facilitated detection of the Fn lipid A major ion (m/z 1665) from extracts of infected mouse spleen and the temperature-modified Fn lipid A at m/z 1637 from infected D. variabilis ticks. Finally, we simultaneously mapped bacterial phospholipid signatures within an Fn infected spleen along with exclusively host-derived inositol-based phospholipid (m/z 933) demonstrating co-profiling for the host-pathogen interaction. Expanded use of NRM matrix in other infection models and endotoxin-targeting imaging experiments will improve our understanding of the lipid interactions at the host-pathogen interface.

Item Type: Journal article
Publication Title: Pathogens and Disease
Creators: Scott, A.J., Flinders, B., Cappell, J., Liang, T., Pelc, R.S., Tran, B., Kilgour, D.P.A., Heeren, R.M.A., Goodlett, D.R. and Ernst, R.K.
Publisher: Oxford University Press
Date: 19 September 2016
ISSN: 2049-632X
Identifiers:
NumberType
10.1093/femspd/ftw097DOI
Divisions: Schools > School of Science and Technology
Depositing User: Jonathan Gallacher
Date Added: 27 Sep 2016 15:56
Last Modified: 13 Oct 2017 15:32
URI: http://irep.ntu.ac.uk/id/eprint/28659

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