{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,2]],"date-time":"2026-06-02T01:32:05Z","timestamp":1780363925830,"version":"3.54.1"},"reference-count":83,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2023,2,21]],"date-time":"2023-02-21T00:00:00Z","timestamp":1676937600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Intramural Research Program of the National Institute of Dental and Craniofacial Research and the Clinical Center, NIH","award":["ZIA CL090034-10"],"award-info":[{"award-number":["ZIA CL090034-10"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biosensors"],"abstract":"<jats:p>Antibody measurements play a central role in the diagnosis of many autoimmune and infectious diseases. One antibody detection technology, Luciferase Immunoprecipitation Systems (LIPS), utilizes genetically encoded recombinant luciferase antigen fusion proteins in an immunoglobulin capture format to generate robust antibody measurement with high diagnostic sensitivity and specificity. The LIPS technology has been highly useful in detecting antibodies for research diagnostics and the discovery of new autoantigens. The methodology of the assay requires immunoglobulin binding reagents such as protein A\/G beads and washing steps to process the immune complex before antibody levels are measured by light production with a luminometer. Recently, simplified mix and read immunoassays based on split components of the nanoluciferase enzyme in a complementation format have been developed for antibody measurements without requiring immunoglobulin-capturing beads or washing steps. The mix and read immunoassays utilize two or three nanoluciferase fragments which when reconstituted via antigen-specific antibody binding generate a functional enzyme. At present, these split luciferase tests have been developed mainly for detecting SARS-CoV-2 antibodies. Here, we describe the traditional LIPS technology and compare it to the new split luciferase methodologies focusing on their technical features, strengths, limitations, and future opportunities for diagnostic research, and clinical applications.<\/jats:p>","DOI":"10.3390\/bios13030303","type":"journal-article","created":{"date-parts":[[2023,2,22]],"date-time":"2023-02-22T02:31:55Z","timestamp":1677033115000},"page":"303","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Advancing Luciferase-Based Antibody Immunoassays to Next-Generation Mix and Read Testing"],"prefix":"10.3390","volume":"13","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1717-048X","authenticated-orcid":false,"given":"Peter D.","family":"Burbelo","sequence":"first","affiliation":[{"name":"Adeno-Associated Virus Biology Section, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 202892, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Youngmi","family":"Ji","sequence":"additional","affiliation":[{"name":"Adeno-Associated Virus Biology Section, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 202892, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael J.","family":"Iadarola","sequence":"additional","affiliation":[{"name":"Department of Perioperative Medicine, Clinical Center, National Institutes of Health, Bethesda, MD 202892, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,2,21]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"e0003521","DOI":"10.1128\/CMR.00035-21","article-title":"The Standardization and Control of Serology and Nucleic Acid Testing for Infectious Diseases","volume":"34","author":"Dimech","year":"2021","journal-title":"Clin. 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