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Open Access Methodology

A mass spectrometric strategy for absolute quantification of Plasmodium falciparum proteins of low abundance

Paul M Southworth12, John E Hyde1 and Paul FG Sims1*

Author Affiliations

1 Manchester Interdisciplinary Biocentre, Faculty of Life Sciences, University of Manchester, 131 Princess Street, Manchester M1 7DN, UK

2 Laboratory of Malaria Immunology and Vaccinology, National Institute of Allergy and Infectious Disease, National Institutes of Health, 12735 Twinbrook Parkway, Rockville MD 20814, USA

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Malaria Journal 2011, 10:315  doi:10.1186/1475-2875-10-315

Published: 25 October 2011

Abstract

Selected reaction monitoring mass spectrometry has been combined with the use of an isotopically labelled synthetic protein, made up of proteotypic tryptic peptides selected from parasite proteins of interest. This allows, for the first time, absolute quantification of proteins from Plasmodium falciparum. This methodology is demonstrated to be of sufficient sensitivity to quantify, even within whole cell extracts, proteins of low abundance from the folate pathway as well as more abundant "housekeeping" proteins.

Keywords:
Absolute quantification of proteins; enzymes of folate metabolism; heavy isotope labelling; malaria parasites; QconCAT