{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,29]],"date-time":"2026-09-29T14:08:28Z","timestamp":1790690908224,"version":"4.1.0"},"reference-count":41,"publisher":"American Chemical Society (ACS)","issue":"38","license":[{"start":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T00:00:00Z","timestamp":1788134400000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T00:00:00Z","timestamp":1788134400000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,8,31]],"date-time":"2026-08-31T00:00:00Z","timestamp":1788134400000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/100014989","name":"Chan Zuckerberg Initiative","doi-asserted-by":"publisher","award":["DAF2023-331951"],"award-info":[{"award-number":["DAF2023-331951"]}],"id":[{"id":"10.13039\/100014989","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"publisher","award":["R35GM124755"],"award-info":[{"award-number":["R35GM124755"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"publisher"}]},{"name":"National Science Foundation (NSF)","award":["2018860"],"award-info":[{"award-number":["2018860"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,9,29]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Fragment-ion signals are often too weak to support confident quantification at trace levels. This challenge is particularly acute for limited-input measurements, including single cells. We introduce hydraplexing, a fragment-readout strategy that installs multiple isobaric barcodes on a single analyte so that barcode-derived fragment ions add in tandem mass spectrometry (MS2). Unlike conventional isobaric labeling that increases sample plexity, hydraplexing multiplexes barcode-release pathways within a single precursor to amplify signal quantitation. Here, hydraplexing is established using a standalone multiamine model analyte, 1,2,4,5-tetraaminobenzene (TAB), bearing 2\u20134 tandem mass tags as TAB-TMT2, TAB-TMT3, and TAB-TMT4 derivatives, which isolates barcode multiplicity without coupling the scaffold to target analytes. Using this controlled system, we mapped barcode-release dissociation networks and defined collision-energy regimes that maximize reporter-ion (RI) output while maintaining precursor transmission during higher-energy collisional dissociation. RI output increased with tag number across direct-infusion electrospray as well as liquid chromatography and capillary electrophoresis separations using parallel-reaction monitoring. Complementary fragment ions (TMTc) peaked at lower collision energies and were more condition-sensitive, identifying RIs as the more robust quantitative channel under the conditions tested. This amplification persisted in a peptide-rich single-cell Xenopus laevis digest matrix, where TAB-TMT4 produced an \u223c3.5-fold higher RI\/precursor response than TAB-TMT2. Hydraplexing was further transferred to 3,4,5-triaminobenzoic acid (TABA), a carboxylate-functionalized scaffold forming 1\u00d7\u20133\u00d7 TMT derivatives, for which the 2\u00d7 and 3\u00d7 derivatives yielded 1.4-fold and 2.7-fold higher RI output than the 1\u00d7 derivative. Hydraplexing establishes fragment-level signal summation as a tunable design principle for engineering additive tag-derived fragment-ion outputs.<\/jats:p>","DOI":"10.1021\/acs.analchem.6c01384","type":"journal-article","created":{"date-parts":[[2026,9,3]],"date-time":"2026-09-03T05:13:11Z","timestamp":1788412391000},"page":"27918-27926","source":"Crossref","is-referenced-by-count":0,"title":["Hydraplexing: Design Principles for Fragment-Level\nSignal Amplification Using a Multi-Amine Model Analyte"],"prefix":"10.1021","volume":"98","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1103-1358","authenticated-orcid":true,"given":"Ailing","family":"Li","sequence":"first","affiliation":[{"name":"University of Maryland , , , ,","place":["College Park, Maryland, United States, 20742"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Navid J.","family":"Ayon","sequence":"additional","affiliation":[{"name":"University of Maryland , , , ,","place":["College Park, Maryland, United States, 20742"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4704-4997","authenticated-orcid":true,"given":"Peter","family":"Nemes","sequence":"additional","affiliation":[{"name":"University of Maryland , , , ,","place":["College Park, Maryland, United States, 20742"]},{"name":"University of Maryland , , , ,","place":["College Park, Maryland, United States, 20742"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,8,31]]},"reference":[{"issue":"28","key":"2026092909093530100_cit1","doi-asserted-by":"publisher","first-page":"6889","DOI":"10.1007\/s00216-023-04759-8","article-title":"A review of the current state of single-cell proteomics\nand future perspective","volume":"415","author":"Ahmad","year":"2023","journal-title":"Anal. Bioanal. Chem."},{"key":"2026092909093530100_cit2","doi-asserted-by":"publisher","first-page":"87","DOI":"10.1007\/978-1-0716-2525-5_5","article-title":"Single-cell\nmass-spectrometry of metabolites\nand proteins for systems and functional biology","volume-title":"Neuromethods","author":"Portero","year":"2022"},{"key":"2026092909093530100_cit3","doi-asserted-by":"publisher","first-page":"106","DOI":"10.1002\/mas.21824","article-title":"Biological\nmass spectrometry enables spatiotemporal \u2019omics: From tissues\nto cells to organelles","volume":"43","author":"Pade","year":"2024","journal-title":"Mass Spectrom. Rev."},{"issue":"3","key":"2026092909093530100_cit4","doi-asserted-by":"publisher","first-page":"363","DOI":"10.1038\/s41592-023-01791-5","article-title":"Single-cell proteomics\nenabled by next-generation sequencing or mass spectrometry","volume":"20","author":"Bennett","year":"2023","journal-title":"Nat. Methods"},{"issue":"13","key":"2026092909093530100_cit5","doi-asserted-by":"publisher","first-page":"dev201492","DOI":"10.1242\/dev.201492","article-title":"Single-cell\nproteomics: quantifying post-transcriptional regulation during development\nwith mass-spectrometry","volume":"150","author":"Slavov","year":"2023","journal-title":"Development"},{"issue":"1-2","key":"2026092909093530100_cit6","doi-asserted-by":"publisher","first-page":"e23012","DOI":"10.1002\/dvg.23012","article-title":"New-generation mass\nspectrometry expands the toolbox of cell and developmental biology","volume":"55","author":"Lombard-Banek","year":"2017","journal-title":"Genesis"},{"key":"2026092909093530100_cit7","doi-asserted-by":"publisher","first-page":"100","DOI":"10.3389\/fcell.2016.00100","article-title":"High-sensitivity\nmass spectrometry\nfor probing gene translation in single embryonic cells in the early\nfrog (Xenopus) embryo","volume":"4","author":"Lombard-Banek","year":"2016","journal-title":"Frontiers\nin Cell and Developmental Biology"},{"issue":"1","key":"2026092909093530100_cit8","doi-asserted-by":"publisher","first-page":"42","DOI":"10.3390\/molecules24010042","article-title":"Recent advances\nand new perspectives in capillary electrophoresis mass spectrometry\nfor single cell omics","volume":"24","author":"DeLaney","year":"2019","journal-title":"Molecules"},{"key":"2026092909093530100_cit9","doi-asserted-by":"publisher","first-page":"149012","DOI":"10.1016\/j.scitotenv.2021.149012","article-title":"Non-targeted\nLC-MS and CE-MS for biomarker discovery in bioreactors: Influence\nof separation, mass spectrometry and data processing tools","volume":"798","author":"Ho\u0308cker","year":"2021","journal-title":"Sci. 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Proteome Res."},{"issue":"11","key":"2026092909093530100_cit30","doi-asserted-by":"publisher","first-page":"5340","DOI":"10.1021\/ac400845e","article-title":"Increasing throughput\nin targeted proteomics assays: 54-plex quantitation in a single mass\nspectrometry run","volume":"85","author":"Everley","year":"2013","journal-title":"Anal. Chem."},{"issue":"12","key":"2026092909093530100_cit31","doi-asserted-by":"publisher","first-page":"5398","DOI":"10.1038\/s41596-021-00616-z","article-title":"Multiplexed\nsingle-cell proteomics\nusing SCoPE2","volume":"16","author":"Petelski","year":"2021","journal-title":"Nat. Protoc."},{"issue":"2","key":"2026092909093530100_cit32","doi-asserted-by":"publisher","first-page":"692","DOI":"10.1021\/acs.jproteome.3c00541","article-title":"Dilute to enrich for\ndeeper proteomics: A yolk-depleted carrier for limited populations\nof embryonic (frog) cells","volume":"23","author":"Pade","year":"2024","journal-title":"J. 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