ArticleNature protocols2015
Complementary IMAC enrichment methods for HLA-associated phosphopeptide identification by mass spectrometry.
Article in Nature protocols, 2015. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 31 papers.
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Who cites it
31 citing papers in PubMed, 81 citations in OpenAlex.
- Strategies for multimodal spatiotemporal profiling of phosphorylation in cilia biology.Journal of cell science · 2025Review
- Mass Spectrometry-Based Proteomics for the Masses: Peptide and Protein Identification in the Hunt Laboratory During the 2000's.Molecular & cellular proteomics : MCP · 2024Article
- Crucial Parameters for Immunopeptidome Characterization: A Systematic Evaluation.International journal of molecular sciences · 2024Article
- Current perspectives on mass spectrometry-based immunopeptidomics: the computational angle to tumor antigen discovery.Journal for immunotherapy of cancer · 2023Review
- Characteristics of Immune Memory and Effector Activity to Cancer-Expressed MHC Class I Phosphopeptides Differ in Healthy Donors and Ovarian Cancer Patients.Cancer immunology research · 2021Article
- Tumor Infiltrating Lymphocytes Target HLA-I Phosphopeptides Derived From Cancer Signaling in Colorectal Cancer.Frontiers in immunology · 2021Article
- Mild Acid Elution and MHC Immunoaffinity Chromatography Reveal Similar Albeit Not Identical Profiles of the HLA Class I Immunopeptidome.Journal of proteome research · 2021Article
- MHC Phosphopeptides: Promising Targets for Immunotherapy of Cancer and Other Chronic Diseases.Molecular & cellular proteomics : MCP · 2021Review
- Critical Review of Existing MHC I Immunopeptidome Isolation Methods.Molecules (Basel, Switzerland) · 2020Review
- Glass Fiber-Supported Hybrid Monolithic Spin Tip for Enrichment of Phosphopeptides from Urinary Extracellular Vesicles.Analytical chemistry · 2020Article
- Towards new horizons: characterization, classification and implications of the tumour antigenic repertoire.Nature reviews. Clinical oncology · 2020Review
- Immunopeptidomic Analysis Reveals That Deamidated HLA-bound Peptides Arise Predominantly from Deglycosylated Precursors.Molecular & cellular proteomics : MCP · 2020Article
- Empirical and Rational Design of T Cell Receptor-Based Immunotherapies.Frontiers in immunology · 2020Review
- Neoantigen-Specific Adoptive Cell Therapies for Cancer: Making T-Cell Products More Personal.Frontiers in immunology · 2020Review
- The Human Immunopeptidome Project: A Roadmap to Predict and Treat Immune Diseases.Molecular & cellular proteomics : MCP · 2020Article
- Murine xenograft bioreactors for human immunopeptidome discovery.Scientific reports · 2019Article
- Mass spectrometry-based identification of MHC-bound peptides for immunopeptidomics.Nature protocols · 2019Article
- Phytic acid functionalized FeMikrochimica acta · 2019Article
- Review
- Minimal Information About an Immuno-Peptidomics Experiment (MIAIPE).Proteomics · 2018Article
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Authors and funding
10 authors at 3 institutions in 2 countries.
Funding
Abstract
Phosphorylation events within cancer cells often become dysregulated, leading to oncogenic signaling and abnormal cell growth. Phosphopeptides derived from aberrantly phosphorylated proteins that are presented on tumors and not on normal tissues by human leukocyte antigen (HLA) class I molecules are promising candidates for future cancer immunotherapies, because they are tumor specific and have been shown to elicit cytotoxic T cell responses. Robust phosphopeptide enrichments that are suitable for low input amounts must be developed to characterize HLA-associated phosphopeptides from clinical samples that are limited by material availability. We present two complementary mass spectrometry-compatible, iron(III)-immobilized metal affinity chromatography (IMAC) methods that use either nitrilotriacetic acid (NTA) or iminodiacetic acid (IDA) in-house-fabricated columns. We developed these protocols to enrich for subfemtomole-level phosphopeptides from cell line and human tissue samples containing picograms of starting material, which is an order of magnitude less material than what is commonly used. In addition, we added a peptide esterification step to increase phosphopeptide specificity from these low-input samples. To date, hundreds of phosphopeptides displayed on melanoma, ovarian cancer, leukemia and colorectal cancer have been identified using these highly selective phosphopeptide enrichment protocols in combination with a program called 'CAD Neutral Loss Finder' that identifies all spectra containing the characteristic neutral loss of phosphoric acid from phosphorylated serine and threonine residues. This methodology enables the identification of HLA-associated phosphopeptides presented by human tissue samples containing as little as nanograms of peptide material in 2 d.
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