{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,24]],"date-time":"2026-10-24T01:58:26Z","timestamp":1792807106912,"version":"4.5.0"},"reference-count":40,"publisher":"Oxford University Press (OUP)","issue":"8","license":[{"start":{"date-parts":[[2023,2,27]],"date-time":"2023-02-27T00:00:00Z","timestamp":1677456000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"funder":[{"DOI":"10.13039\/100006492","name":"Division of Intramural Research, National Institute of Allergy and Infectious Diseases","doi-asserted-by":"publisher","award":["R01AI122912"],"award-info":[{"award-number":["R01AI122912"]}],"id":[{"id":"10.13039\/100006492","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000057","name":"HHS | NIH | National Institute of General Medical Sciences","doi-asserted-by":"publisher","award":["T32GM095421"],"award-info":[{"award-number":["T32GM095421"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000057","name":"HHS | NIH | National Institute of General Medical Sciences","doi-asserted-by":"publisher","award":["P20GM113132"],"award-info":[{"award-number":["P20GM113132"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100016127","name":"Fast Grants","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100016127","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Fred Hutchinson Cancer Center Covid Pilot Award","id":[{"id":"https:\/\/ror.org\/007ps6h72","id-type":"ROR","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2023,4,15]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>The study of Ag-specific lymphocytes has been a key advancement in immunology over the past few decades. The development of multimerized probes containing Ags, peptide:MHC complexes, or other ligands was one innovation allowing the direct study of Ag-specific lymphocytes by flow cytometry. Although these types of study are now common and performed by thousands of laboratories, quality control and assessment of probe quality are often minimal. In fact, many of these types of probe are made in-house, and protocols vary between laboratories. Although peptide:MHC multimers can often be obtained from commercial sources or core facilities, few such services exist for Ag multimers. To ensure high quality and consistency with ligand probes, we have developed an easy and robust multiplexed approach using commercially available beads able to bind Abs specific for the ligand of interest. Using this assay, we have sensitively assessed the performance of peptide:MHC and Ag tetramers and have found considerable batch-to-batch variability in performance and stability over time more easily than using murine or human cell-based assays. This bead-based assay can also reveal common production errors such as miscalculation of Ag concentration. This work could set the stage for the development of standardized assays for all commonly used ligand probes to limit laboratory-to-laboratory technical variation and experimental failure caused by probe underperformance.<\/jats:p>","DOI":"10.4049\/jimmunol.2200934","type":"journal-article","created":{"date-parts":[[2023,3,8]],"date-time":"2023-03-08T09:11:27Z","timestamp":1678266687000},"page":"1156-1165","source":"Crossref","is-referenced-by-count":14,"title":["Validation of Ligand Tetramers for the Detection of Antigen-Specific Lymphocytes"],"prefix":"10.1093","volume":"210","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8086-5239","authenticated-orcid":false,"given":"Kristin S","family":"Fitzpatrick","sequence":"first","affiliation":[{"name":"Immunology and Vaccine Development Program, Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Center , Seattle, WA"},{"name":"Molecular Medicine and Mechanisms of Disease PhD Program, University of Washington , Seattle, WA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hanna N","family":"Degefu","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth College , Lebanon, NH"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katrina","family":"Poljakov","sequence":"additional","affiliation":[{"name":"Immunology and Vaccine Development Program, Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Center , Seattle, WA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Madeleine G","family":"Bibby","sequence":"additional","affiliation":[{"name":"Immunology and Vaccine Development Program, Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Center , Seattle, WA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5155-4116","authenticated-orcid":false,"given":"Allison J","family":"Remington","sequence":"additional","affiliation":[{"name":"Immunology and Vaccine Development Program, Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Center , Seattle, WA"},{"name":"Division of Dermatology, Department of Medicine, University of Washington , Seattle, WA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3146-4812","authenticated-orcid":false,"given":"Tyler G","family":"Searles","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth College , Lebanon, NH"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4260-9784","authenticated-orcid":false,"given":"Matthew D","family":"Gray","sequence":"additional","affiliation":[{"name":"Immunology and Vaccine Development Program, Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Center , Seattle, WA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5858-2662","authenticated-orcid":false,"given":"Jim","family":"Boonyaratanakornkit","sequence":"additional","affiliation":[{"name":"Immunology and Vaccine Development Program, Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Center , Seattle, WA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7909-3868","authenticated-orcid":false,"given":"Pamela C","family":"Rosato","sequence":"additional","affiliation":[{"name":"Department of Microbiology and Immunology, Geisel School of Medicine at Dartmouth College , Lebanon, NH"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9718-2901","authenticated-orcid":false,"given":"Justin J","family":"Taylor","sequence":"additional","affiliation":[{"name":"Immunology and Vaccine Development Program, Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Center , Seattle, WA"},{"name":"Department of Immunology, University of Washington , Seattle, WA"},{"name":"Department of Global Health, University of Washington , Seattle, WA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2023,2,27]]},"reference":[{"key":"2025021008031023100_r1","doi-asserted-by":"crossref","first-page":"1379","DOI":"10.1073\/pnas.84.5.1379","article-title":"Isolation of high-affinity memory B cells: phycoerythrin as a probe for antigen-binding cells","volume":"84","author":"Hayakawa","year":"1987","journal-title":"Proc. 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