RT Journal Article
T1 Shotgun Characterization of the Circulating IgM Antigenome of an Infectious Pathogen by Immunocapture-LC–MS/MS from Dried Serum Spots
A1 Abad González, Paloma
A1 Coronado Brieva, Montserrat
A1 Vincelle-Nieto, África
A1 Pérez Benavente, Susana
A1 Fobil, Julius N.
A1 Puyet Catalina, Antonio
A1 Díez Martín, Amalia
A1 Reyes Palomares, Armando Adolfo
A1 Azcárate, Isabel G.
A1 Bautista Santa Cruz, José Manuel
AB 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.
PB American Chemical Society
SN 1535-3893
YR 2024
FD 2024-01-06
LK https://hdl.handle.net/20.500.14352/103791
UL https://hdl.handle.net/20.500.14352/103791
LA eng
NO Paloma 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.3c00439
DS Docta Complutense
RD 8 abr 2025