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Guidance document: validation of a high-performance liquid chromatography-tandem mass spectrometry immunopeptidomics assay for the identification of HLA class I ligands suitable for pharmaceutical therapies.

Abstract

For more than two decades naturally presented, human leukocyte antigen (HLA)-restricted peptides (immunopeptidome) have been eluted and sequenced using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Since, identified disease-associated HLA ligands have been characterized and evaluated as potential active substances. Treatments based on HLA-presented peptides have shown promising results in clinical application as personalized T cell-based immunotherapy. Peptide vaccination cocktails are produced as investigational medicinal products under GMP conditions. In order to support clinical trials based on HLA-presented tumor-associated antigens, in this study the sensitive LC-MS/MS HLA class I antigen identification pipeline was fully validated for our technical equipment according to the current US Food and Drug Administration (FDA) and European Medicines Agency (EMA) guidelines.The immunopeptidomes of JY cells with or without spiked-in, isotope labeled peptides, of peripheral blood mononuclear cells of healthy volunteers as well as a chronic lymphocytic leukemia and a bladder cancer sample were reliably identified using a data-dependent acquisition method. As the LC-MS/MS pipeline is used for identification purposes, the validation parameters include accuracy, precision, specificity, limit of detection and robustness.

Authors+Show Affiliations

Department of Immunology, Interfaculty Institute for Cell Biology, Germany.Interfaculty Institute for Cell Biology, Germany.Interfaculty Institute for Cell Biology, Germany.Interfaculty Institute for Cell Biology, Germany.Interfaculty Institute for Cell Biology, Germany.University of Tubingen.University of Tuebingen.University of T�bingen, Germany stefan.stevanovic@uni-tuebingen.de.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

31937595

Citation

Ghosh, Michael, et al. "Guidance Document: Validation of a High-performance Liquid Chromatography-tandem Mass Spectrometry Immunopeptidomics Assay for the Identification of HLA Class I Ligands Suitable for Pharmaceutical Therapies." Molecular & Cellular Proteomics : MCP, 2020.
Ghosh M, Gauger M, Marcu A, et al. Guidance document: validation of a high-performance liquid chromatography-tandem mass spectrometry immunopeptidomics assay for the identification of HLA class I ligands suitable for pharmaceutical therapies. Mol Cell Proteomics. 2020.
Ghosh, M., Gauger, M., Marcu, A., Nelde, A., Denk, M., Schuster, H., ... Stevanovic, S. (2020). Guidance document: validation of a high-performance liquid chromatography-tandem mass spectrometry immunopeptidomics assay for the identification of HLA class I ligands suitable for pharmaceutical therapies. Molecular & Cellular Proteomics : MCP, doi:10.1074/mcp.C119.001652.
Ghosh M, et al. Guidance Document: Validation of a High-performance Liquid Chromatography-tandem Mass Spectrometry Immunopeptidomics Assay for the Identification of HLA Class I Ligands Suitable for Pharmaceutical Therapies. Mol Cell Proteomics. 2020 Jan 14; PubMed PMID: 31937595.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Guidance document: validation of a high-performance liquid chromatography-tandem mass spectrometry immunopeptidomics assay for the identification of HLA class I ligands suitable for pharmaceutical therapies. AU - Ghosh,Michael, AU - Gauger,Marion, AU - Marcu,Ana, AU - Nelde,Annika, AU - Denk,Monika, AU - Schuster,Heiko, AU - Rammensee,Hans Georg, AU - Stevanovic,Stefan, Y1 - 2020/01/14/ PY - 2020/01/14/accepted PY - 2019/07/01/received PY - 2020/01/09/revised PY - 2020/1/16/entrez KW - Cancer therapeutics KW - GMP KW - Immunopeptidome KW - Individualized medicine* KW - LC–MS/MS KW - Mass Spectrometry KW - Peptidomics KW - Personalized medicine KW - Validation JF - Molecular & cellular proteomics : MCP JO - Mol. Cell Proteomics N2 - For more than two decades naturally presented, human leukocyte antigen (HLA)-restricted peptides (immunopeptidome) have been eluted and sequenced using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Since, identified disease-associated HLA ligands have been characterized and evaluated as potential active substances. Treatments based on HLA-presented peptides have shown promising results in clinical application as personalized T cell-based immunotherapy. Peptide vaccination cocktails are produced as investigational medicinal products under GMP conditions. In order to support clinical trials based on HLA-presented tumor-associated antigens, in this study the sensitive LC-MS/MS HLA class I antigen identification pipeline was fully validated for our technical equipment according to the current US Food and Drug Administration (FDA) and European Medicines Agency (EMA) guidelines.The immunopeptidomes of JY cells with or without spiked-in, isotope labeled peptides, of peripheral blood mononuclear cells of healthy volunteers as well as a chronic lymphocytic leukemia and a bladder cancer sample were reliably identified using a data-dependent acquisition method. As the LC-MS/MS pipeline is used for identification purposes, the validation parameters include accuracy, precision, specificity, limit of detection and robustness. SN - 1535-9484 UR - https://www.unboundmedicine.com/medline/citation/31937595/Guidance_document:_validation_of_a_high-performance_liquid_chromatography-tandem_mass_spectrometry_immunopeptidomics_assay_for_the_identification_of_HLA_class_I_ligands_suitable_for_pharmaceutical_therapies L2 - http://www.mcponline.org/cgi/pmidlookup?view=long&pmid=31937595 DB - PRIME DP - Unbound Medicine ER -