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Dissection of schistosome tissues under LC–MS compatible preservative conditions for quantitative proteomics.

Authors :
Neves, Leandro Xavier
Wilson, R. Alan
Brownridge, Philip
Holman, Stephen W.
Harman, Victoria M.
Eyers, Claire E.
Beynon, Robert J.
Castro‐Borges, William
Source :
Rapid Communications in Mass Spectrometry: RCM. Jan2024 Supplement 1, Vol. 38, p1-15. 15p.
Publication Year :
2024

Abstract

Schistosomes are blood flukes with specialised tissues and organs, each one playing a pivotal role in perpetuating the parasite life cycle. Herein, we describe a detailed methodology for preserving the proteome of adult Schistosoma mansoni worms during manual dissection for enrichment of tissues associated with the parasite's alimentary tract. We provide step‐by‐step directions for specimen storage and dissection while in preservative solution, tissue homogenisation, protein extraction and digestion using a methodology fully compatible with downstream quantitative liquid chromatography–mass spectrometry analysis. Our methodology uses label‐free and QconCAT‐based absolute quantification for detection of S. mansoni oesophageal gland products proposed as vaccine candidates. Through stabilisation of the proteome and minimising sample degradation during dissection our approach has allowed us to access the hidden proteome of target tissues not readily available from total lysates because of their small volume. This protocol can be replicated or adapted to other Schistosoma species lacking quantitative proteomics characterisation of specialised tissues for discovery of proteins with potential diagnostic and therapeutic utility. [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
09514198
Volume :
38
Database :
Academic Search Index
Journal :
Rapid Communications in Mass Spectrometry: RCM
Publication Type :
Academic Journal
Accession number :
175853496
Full Text :
https://doi.org/10.1002/rcm.9523