Advancing Luciferase-Based Antibody Immunoassays to Next-Generation Mix and Read Testing
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...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-02-01
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| Series: | Biosensors |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2079-6374/13/3/303 |
| _version_ | 1797613228225724416 |
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| author | Peter D. Burbelo Youngmi Ji Michael J. Iadarola |
| author_facet | Peter D. Burbelo Youngmi Ji Michael J. Iadarola |
| author_sort | Peter D. Burbelo |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-11T06:52:55Z |
| format | Article |
| id | doaj.art-c47fa9aa60e64447aa592983bd959eda |
| institution | Directory Open Access Journal |
| issn | 2079-6374 |
| language | English |
| last_indexed | 2024-03-11T06:52:55Z |
| publishDate | 2023-02-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biosensors |
| spelling | doaj.art-c47fa9aa60e64447aa592983bd959eda2023-11-17T09:53:28ZengMDPI AGBiosensors2079-63742023-02-0113330310.3390/bios13030303Advancing Luciferase-Based Antibody Immunoassays to Next-Generation Mix and Read TestingPeter D. Burbelo0Youngmi Ji1Michael J. Iadarola2Adeno-Associated Virus Biology Section, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 202892, USAAdeno-Associated Virus Biology Section, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, MD 202892, USADepartment of Perioperative Medicine, Clinical Center, National Institutes of Health, Bethesda, MD 202892, USAAntibody 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.https://www.mdpi.com/2079-6374/13/3/303antibodyautoantibodyLIPSluciferase-based immunoassayssplit luciferase |
| spellingShingle | Peter D. Burbelo Youngmi Ji Michael J. Iadarola Advancing Luciferase-Based Antibody Immunoassays to Next-Generation Mix and Read Testing Biosensors antibody autoantibody LIPS luciferase-based immunoassays split luciferase |
| title | Advancing Luciferase-Based Antibody Immunoassays to Next-Generation Mix and Read Testing |
| title_full | Advancing Luciferase-Based Antibody Immunoassays to Next-Generation Mix and Read Testing |
| title_fullStr | Advancing Luciferase-Based Antibody Immunoassays to Next-Generation Mix and Read Testing |
| title_full_unstemmed | Advancing Luciferase-Based Antibody Immunoassays to Next-Generation Mix and Read Testing |
| title_short | Advancing Luciferase-Based Antibody Immunoassays to Next-Generation Mix and Read Testing |
| title_sort | advancing luciferase based antibody immunoassays to next generation mix and read testing |
| topic | antibody autoantibody LIPS luciferase-based immunoassays split luciferase |
| url | https://www.mdpi.com/2079-6374/13/3/303 |
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