ArticleNature protocols2018
Metal-isotope-tagged monoclonal antibodies for high-dimensional mass cytometry.
Article in Nature protocols, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 135 papers, 1 of them a synthesis that pooled it.
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Who cites it
135 citing papers in PubMed, 1 synthesis or guideline pooled it, 263 citations in OpenAlex.
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- Mass Cytometry Reveals Cell Type-specific Immune Remodeling in Placentas of Women with Chronic Hepatitis B Infection.Journal of clinical and translational hepatology · 2026Article
- AURKB-mediated phosphorylation of metabolic enzyme CD73 drives immune suppression and renal cell carcinoma progression.Cell death and differentiation · 2026Article
- Protocol for single-cell epigenetic profiling in human organoids and tumoroids with Epi-CyTOF.STAR protocols · 2026Article
- Engineering Cathepsin S Selective Chemical Probes and Antibody-Drug Conjugates through Substrate Profiling with Unnatural Amino Acids.Journal of medicinal chemistry · 2026Article
- SHP1 expression in tumor-associated dendritic cells drives immunoevasion via impairing CD8Oncogenesis · 2026Article
- Epstein-Barr virus orchestrates spatial reorganization and immunomodulation in the classic Hodgkin lymphoma tumor microenvironment.Cell reports. Medicine · 2026Article
- Article
- The use of mass cytometry (CyTOF) to evaluate the cellular uptake of stable radiopharmaceutical surrogates in single cells: a proof-of-concept study.EJNMMI radiopharmacy and chemistry · 2026Article
- Single-cell spatial atlas of the aging human breast.Nature aging · 2026Article
- CyTOF analysis of immune characteristics in cSLE: belimumab treatment and refractory cases.Frontiers in immunology · 2026Article
- Unveiling the immune landscape and microenvironmental architecture of IgA nephropathy using single-cell imaging mass cytometry.Clinical & translational immunology · 2026Article
- Sequential versus Simultaneous Quantitative Analysis of Biomarkers in Individual Cells by ICP-MS and Mass Cytometry: A Focus on Immunotherapy.Analytical chemistry · 2025Article
- Elemental mass spectrometry imaging of biomolecules using metal-conjugated probes.Nature reviews. Chemistry · 2025Review
- Single-cell spatial proteomics.Histology and histopathology · 2025Review
- MALDI mass spectrometry imaging of extracellular matrix proteins.The Analyst · 2025Review
- Polymer-Based Mass Cytometry Reagents: Synthesis and Biomedical Applications.Molecules (Basel, Switzerland) · 2025Review
- CyTOF as a suitable tool for stratification and monitoring of cancer patients.Journal of translational medicine · 2025Review
75 more citing papers are in PubMed but not listed here.
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Authors and funding
5 authors at 3 institutions in 1 country.
Funding
Abstract
Advances in single-cell mass cytometry have increasingly improved highly multidimensional characterization of immune cell heterogeneity. The immunoassay multiplexing capacity relies on monoclonal antibodies labeled with stable heavy-metal isotopes. To date, a variety of rare-earth elements and noble and post-transition metal isotopes have been used in mass cytometry; nevertheless, the methods used for antibody conjugation differ because of the individual metal coordination chemistries and distinct stabilities of various metal cations. Herein, we provide three optimized protocols for conjugating monoclonal IgG antibodies with 48 high-purity heavy-metal isotopes: (i) 38 isotopes of lanthanides, 2 isotopes of indium, and 1 isotope of yttrium; (ii) 6 isotopes of palladium; and (iii) 1 isotope of bismuth. Bifunctional chelating agents containing coordinative ligands of monomeric DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid) or polymeric pentetic acid (DTPA) were used to stably sequester isotopic cations in aqueous solutions and were subsequently coupled to IgG antibodies using site-specific biorthogonal reactions. Furthermore, quantification methods based on antibody inherent absorption at 280 nm and on extrinsic absorption at 562 nm after staining with bicinchoninic acid (BCA) are reported to determine metal-isotope-tagged antibodies. In addition, a freeze-drying procedure to prepare palladium isotopic mass tags is described. To demonstrate the utility, experiments using six palladium-tagged CD45 antibodies for barcoding assays of live immune cells in cytometry by time-of-flight (CyTOF) are described. Conjugation of pure isotopes of lanthanides, indium, or yttrium takes ~3.5 h. Conjugation of bismuth takes ~4 h. Preparation of palladium mass tags takes ~8 h. Conjugation of pure isotopes of palladium takes ~2.5 h. Antibody titration takes ~4 h.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.