Shotgun Characterization of the Circulating IgM Antigenome of an Infectious Pathogen by Immunocapture-LC-MS/MS from Dried Serum Spots.

Autor:	Abad P; Department of Biochemistry and Molecular Biology, Universidad Complutense de Madrid, Faculty of Veterinary Sciences, 28040 Madrid, Spain., Coronado M; Department of Biochemistry and Molecular Biology, Universidad Complutense de Madrid, Faculty of Veterinary Sciences, 28040 Madrid, Spain., Vincelle-Nieto Á; Department of Biochemistry and Molecular Biology, Universidad Complutense de Madrid, Faculty of Veterinary Sciences, 28040 Madrid, Spain., Pérez-Benavente S; Department of Biochemistry and Molecular Biology, Universidad Complutense de Madrid, Faculty of Veterinary Sciences, 28040 Madrid, Spain., Fobil JN; Department of Biological, Environmental & Occupational Health Sciences, School of Public Health, College of Health Sciences, University of Ghana, P.O. Box LG 13, G-4381 Legon, Ghana., Puyet A; Department of Biochemistry and Molecular Biology, Universidad Complutense de Madrid, Faculty of Veterinary Sciences, 28040 Madrid, Spain.; 12 de Octubre Health Research Institute (imas12), 28041 Madrid, Spain., Diez A; Department of Biochemistry and Molecular Biology, Universidad Complutense de Madrid, Faculty of Veterinary Sciences, 28040 Madrid, Spain.; 12 de Octubre Health Research Institute (imas12), 28041 Madrid, Spain., Reyes-Palomares A; Department of Biochemistry and Molecular Biology, Universidad Complutense de Madrid, Faculty of Veterinary Sciences, 28040 Madrid, Spain.; 12 de Octubre Health Research Institute (imas12), 28041 Madrid, Spain., Azcárate IG; Faculty of Health Sciences, Rey Juan Carlos University, Alcorcón, 28922 Madrid, Spain., Bautista JM; Department of Biochemistry and Molecular Biology, Universidad Complutense de Madrid, Faculty of Veterinary Sciences, 28040 Madrid, Spain.; 12 de Octubre Health Research Institute (imas12), 28041 Madrid, Spain.
Jazyk:	angličtina
Zdroj:	Journal of proteome research [J Proteome Res] 2024 Feb 02; Vol. 23 (2), pp. 633-643. Date of Electronic Publication: 2024 Jan 06.
DOI:	10.1021/acs.jproteome.3c00439
Abstrakt:	One of the main challenges in compiling the complete collection of protein antigens from pathogens for the selection of vaccine candidates or intervention targets is to acquire a broad enough representation of them to be recognized by the highly diversified immunoglobulin repertoire in human populations. Dried serum spot sampling (DSS) retains a large repertoire of circulating immunoglobulins from each individual that can be representative of a population, according to the sample size. In this work, shotgun proteomics of an infectious pathogen based on DSS sampling coupled with IgM immunoprecipitation, liquid chromatography-mass spectrometry (LC-MS/MS), and bioinformatic analyses was combined to characterize the circulating IgM antigenome. Serum samples from a malaria endemic region at different clinical statuses were studied to optimize IgM binding efficiency and antibody leaching by varying serum/immunomagnetic bead ratios and elution conditions. The method was validated using Plasmodium falciparum extracts identifying 110 of its IgM-reactive antigens while minimizing the presence of human proteins and antibodies. Furthermore, the IgM antigen recognition profile differentiated between malaria-infected and noninfected individuals at the time of sampling. We conclude that a shotgun proteomics approach offers advantages in providing a high-throughput, reliable, and clean way to identify IgM-recognized antigens from trace amounts of serum. The mass spectrometry raw data and metadata have been deposited with ProteomeXchange via MassIVE with the PXD identifier PXD043800.
Databáze:	MEDLINE
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