Abad González, PalomaCoronado Brieva, MontserratVincelle-Nieto, ÁfricaPérez Benavente, SusanaFobil, Julius N.Puyet Catalina, AntonioDíez Martín, AmaliaReyes Palomares, Armando AdolfoAzcárate, Isabel G.Bautista Santa Cruz, José Manuel2024-05-072024-05-072024-01-06Paloma Abad, Montserrat Coronado, África Vincelle-Nieto, Susana Pérez-Benavente, Julius N. Fobil, Antonio Puyet, Amalia Diez, Armando Reyes-Palomares, Isabel G. Azcárate, and José M. Bautista. Journal of Proteome Research 2024 23 (2), 633-643 DOI: 10.1021/acs.jproteome.3c004391535-389310.1021/acs.jproteome.3c00439https://hdl.handle.net/20.500.14352/103791One 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.engAttribution-NonCommercial-NoDerivatives 4.0 Internationalhttp://creativecommons.org/licenses/by-nc-nd/4.0/journal article1535-3907https://doi.org/10.1021/acs.jproteome.3c0043938183416https://pubmed.ncbi.nlm.nih.gov/38183416/embargoed access577.1612.015612.017ImmunoglobulinImmunomicsMalariaAntigensInfectionImmunoprecipitationCiencias Biomédicas2403 Bioquímica