<?xml version="1.0" encoding="UTF-8" standalone="yes"?> <ProteomeXchangeDataset id="PXD028633" formatVersion="1.4.0" xsi:noNamespaceSchemaLocation="proteomeXchange-1.4.0.xsd" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance"> <CvList> <Cv fullName="PSI-MS" uri="https://raw.githubusercontent.com/HUPO-PSI/psi-ms-CV/master/psi-ms.obo" id="MS"/> <Cv fullName="PSI-MOD" uri="https://raw.githubusercontent.com/MICommunity/psidev/master/psi/mod/data/PSI-MOD.obo" id="MOD"/> <Cv fullName="UNIMOD" uri="http://www.unimod.org/obo/unimod.obo" id="UNIMOD"/> </CvList> <ChangeLog> <ChangeLogEntry date="2021-12-08">Updated publication reference for PubMed record(s): 34798331.</ChangeLogEntry> </ChangeLog> <DatasetSummary announceDate="2021-12-07" hostingRepository="PRIDE" title="Isolation of mouse and human MHC class I- and II-associated peptides for the analysis of immunopeptidomes by mass spectrometry"> <Description>Immunopeptidomics is an emerging field that fuels and guides the development of vaccines and immunotherapies. More specifically, it refers to the science of investigating the composition of peptides presented by major histocompatibility complex (MHC) class I and class II molecules using mass spectrometry (MS) technology platforms. Among all the steps in a MS-based immunopeptidomic workflow, sample preparationis critically important for capturing high-quality data of therapeutic relevance. Here, we describe step-by-step instructions to isolate MHC class I and II-associated peptides by immunoaffinity purification from quality control (QC) samples. Our QC samples are MHC class I and class II peptides isolated from mouse (EL4 and A20) and human (JY) cell lines. We thoroughly describe the various reagents and specfic antibodies that are used too isolate MHC-associated peptides from these cell lines, including the steps to verify the beads-binding efficiency of the antibody and the elution efficiency of the MHC-peptide complexes from the beads. QC samples generated using the described protocol can be used to establish and standardize immunopeptidomic workflow, as well as to benchmark new protocols. We anticipate that the visualized protocol provided in this report will represent a great starting point for any non-expert in addition to foster the instra- and inter-laboratory reproducibility of the sample preparation procedure in immunopeptidomics.</Description> <ReviewLevel> <cvParam cvRef="MS" accession="MS:1002854" name="Peer-reviewed dataset"/> </ReviewLevel> <RepositorySupport> <cvParam cvRef="MS" accession="MS:1002856" name="Supported dataset by repository"/> </RepositorySupport> </DatasetSummary> <DatasetIdentifierList> <DatasetIdentifier> <cvParam cvRef="MS" accession="MS:1001919" name="ProteomeXchange accession number" value="PXD028633"/> <cvParam cvRef="MS" accession="MS:1001921" name="ProteomeXchange accession number version number" value="2"/> </DatasetIdentifier> <DatasetIdentifier> <cvParam cvRef="MS" accession="MS:1001922" name="Digital Object Identifier (DOI)" value="10.6019/PXD028633"/> </DatasetIdentifier> </DatasetIdentifierList> <DatasetOriginList> <DatasetOrigin> <cvParam cvRef="MS" accession="MS:1002868" name="Original data"/> </DatasetOrigin> </DatasetOriginList> <SpeciesList> <Species> <cvParam cvRef="MS" accession="MS:1001469" name="taxonomy: scientific name" value="Mus musculus (Mouse)"/> <cvParam cvRef="MS" accession="MS:1001467" name="taxonomy: NCBI TaxID" value="10090"/> </Species> <Species> <cvParam cvRef="MS" accession="MS:1001469" name="taxonomy: scientific name" value="Homo sapiens (Human)"/> <cvParam cvRef="MS" accession="MS:1001467" name="taxonomy: NCBI TaxID" value="9606"/> </Species> </SpeciesList> <InstrumentList> <Instrument id="Instrument_1"> <cvParam cvRef="MS" accession="MS:1003029" name="Orbitrap Eclipse"/> </Instrument> </InstrumentList> <ModificationList> <cvParam cvRef="MOD" accession="MOD:00425" name="monohydroxylated residue"/> <cvParam cvRef="MOD" accession="MOD:00400" name="deamidated residue"/> </ModificationList> <ContactList> <Contact id="project_submitter"> <cvParam cvRef="MS" accession="MS:1000586" name="contact name" value="isabelle sirois"/> <cvParam cvRef="MS" accession="MS:1000589" name="contact email" value="isabelle.sirois.phd@gmail.com"/> <cvParam cvRef="MS" accession="MS:1000590" name="contact affiliation" value="Centre de recherche Hôpital Sainte-Justine"/> <cvParam cvRef="MS" accession="MS:1002037" name="dataset submitter"/> </Contact> <Contact id="project_lab_head"> <cvParam cvRef="MS" accession="MS:1002332" name="lab head"/> <cvParam cvRef="MS" accession="MS:1000586" name="contact name" value="Etienne Caron"/> <cvParam cvRef="MS" accession="MS:1000589" name="contact email" value="Etienne.caron@umontral.ca"/> <cvParam cvRef="MS" accession="MS:1000590" name="contact affiliation" value="1CHU Sainte-Justine Research Center, Montreal, QC H3T 1C5, Canada 2 Department of Pathology and cellular biology, Faculty of Medicine, Universite de Montreal, QC H3T 1J4, Canada"/> </Contact> </ContactList> <PublicationList> <Publication id="PMID34798331"> <cvParam cvRef="MS" accession="MS:1000879" name="PubMed identifier" value="34798331"/> <cvParam cvRef="MS" accession="MS:1002866" name="Reference" value="Kovalchik KA, Ma Q, Wessling L, Saab F, Despault J, Kubiniok P, Hamelin DJ, Faridi P, Li C, Purcell AW, Jang A, Paramithiotis E, Tognetti M, Reiter L, Bruderer R, Lanoix J, Bonneil É, Courcelles M, Thibault P, Caron E, Sirois I. MhcVizPipe: A Quality Control Software for Rapid Assessment of Small- to Large-Scale Immunopeptidome Data Sets. Mol Cell Proteomics. 2021:100178"/> </Publication> </PublicationList> <KeywordList> <cvParam cvRef="MS" accession="MS:1001925" name="submitter keyword" value="immunopeptidome, mass spectrometry, MHC class I associated peptides, MHC class II associated peptides, mouse, Human"/> </KeywordList> <FullDatasetLinkList> <FullDatasetLink> <cvParam cvRef="MS" accession="MS:1002852" name="Dataset FTP location" value="ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2021/12/PXD028633"/> </FullDatasetLink> <FullDatasetLink> <cvParam cvRef="MS" accession="MS:1001930" name="PRIDE project URI" value="http://www.ebi.ac.uk/pride/archive/projects/PXD028633"/> </FullDatasetLink> </FullDatasetLinkList> <DatasetFileList> <DatasetFile id="FILE_223" name="EL4_IPMHCI_1_12MAY2021_R1.mgf"> <cvParam cvRef="MS" accession="MS:1002850" name="Peak list file URI" value="ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2021/12/PXD028633/EL4_IPMHCI_1_12MAY2021_R1.mgf"/> </DatasetFile> <DatasetFile id="FILE_224" 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