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A New Luminescence Assay for Autoantibodies to Mammalian Cell–Prepared Insulinoma-Associated Protein 2
OBJECTIVE—Insulinoma-associated protein 2 (IA-2) is a major autoantigen in type 1 diabetes, and IA-2 autoantibodies are routinely detected by a liquid-phase radioimmunoprecipitation assay. The present experiments were initiated to develop a new assay that does not require the use of radioisotopes or...
Autores principales: | , , , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
American Diabetes Association
2008
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2518352/ https://www.ncbi.nlm.nih.gov/pubmed/18535195 http://dx.doi.org/10.2337/dc08-0286 |
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author | Burbelo, Peter D. Hirai, Hiroki Leahy, Hannah Lernmark, Ake Ivarsson, Sten A. Iadarola, Michael J. Louis Notkins, Abner |
author_facet | Burbelo, Peter D. Hirai, Hiroki Leahy, Hannah Lernmark, Ake Ivarsson, Sten A. Iadarola, Michael J. Louis Notkins, Abner |
author_sort | Burbelo, Peter D. |
collection | PubMed |
description | OBJECTIVE—Insulinoma-associated protein 2 (IA-2) is a major autoantigen in type 1 diabetes, and IA-2 autoantibodies are routinely detected by a liquid-phase radioimmunoprecipitation assay. The present experiments were initiated to develop a new assay that does not require the use of radioisotopes or autoantigens prepared in bacteria or by in vitro transcription/translation. RESEARCH DESIGN AND METHODS—IA-2 luciferase fusion protein was expressed in mammalian cells and assayed for autoantibodies by liquid-phase luciferase immunoprecipitation. RESULTS—Our study showed that there was no significant difference between the luciferase immunoprecipitation and the radioimmunoprecipitation assays in sensitivity and specificity, and comparison of the two assays revealed a high correlation coefficient (R(2) = 0.805). CONCLUSIONS—The luciferase system offers a robust, inexpensive, nonradioactive method for the detection of autoantibodies to mammalian cell–prepared IA-2 and could be of practical value at the clinical level. |
format | Text |
id | pubmed-2518352 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2008 |
publisher | American Diabetes Association |
record_format | MEDLINE/PubMed |
spelling | pubmed-25183522009-09-01 A New Luminescence Assay for Autoantibodies to Mammalian Cell–Prepared Insulinoma-Associated Protein 2 Burbelo, Peter D. Hirai, Hiroki Leahy, Hannah Lernmark, Ake Ivarsson, Sten A. Iadarola, Michael J. Louis Notkins, Abner Diabetes Care Emerging Treatments and Technologies OBJECTIVE—Insulinoma-associated protein 2 (IA-2) is a major autoantigen in type 1 diabetes, and IA-2 autoantibodies are routinely detected by a liquid-phase radioimmunoprecipitation assay. The present experiments were initiated to develop a new assay that does not require the use of radioisotopes or autoantigens prepared in bacteria or by in vitro transcription/translation. RESEARCH DESIGN AND METHODS—IA-2 luciferase fusion protein was expressed in mammalian cells and assayed for autoantibodies by liquid-phase luciferase immunoprecipitation. RESULTS—Our study showed that there was no significant difference between the luciferase immunoprecipitation and the radioimmunoprecipitation assays in sensitivity and specificity, and comparison of the two assays revealed a high correlation coefficient (R(2) = 0.805). CONCLUSIONS—The luciferase system offers a robust, inexpensive, nonradioactive method for the detection of autoantibodies to mammalian cell–prepared IA-2 and could be of practical value at the clinical level. American Diabetes Association 2008-09 /pmc/articles/PMC2518352/ /pubmed/18535195 http://dx.doi.org/10.2337/dc08-0286 Text en Copyright © 2008, DIABETES CARE Readers may use this article as long as the work is properly cited, the use is educational and not for profit, and the work is not altered. See http://creativecommons.org/licenses/by-nc-nd/3.0/ for details. |
spellingShingle | Emerging Treatments and Technologies Burbelo, Peter D. Hirai, Hiroki Leahy, Hannah Lernmark, Ake Ivarsson, Sten A. Iadarola, Michael J. Louis Notkins, Abner A New Luminescence Assay for Autoantibodies to Mammalian Cell–Prepared Insulinoma-Associated Protein 2 |
title | A New Luminescence Assay for Autoantibodies to Mammalian Cell–Prepared Insulinoma-Associated Protein 2 |
title_full | A New Luminescence Assay for Autoantibodies to Mammalian Cell–Prepared Insulinoma-Associated Protein 2 |
title_fullStr | A New Luminescence Assay for Autoantibodies to Mammalian Cell–Prepared Insulinoma-Associated Protein 2 |
title_full_unstemmed | A New Luminescence Assay for Autoantibodies to Mammalian Cell–Prepared Insulinoma-Associated Protein 2 |
title_short | A New Luminescence Assay for Autoantibodies to Mammalian Cell–Prepared Insulinoma-Associated Protein 2 |
title_sort | new luminescence assay for autoantibodies to mammalian cell–prepared insulinoma-associated protein 2 |
topic | Emerging Treatments and Technologies |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2518352/ https://www.ncbi.nlm.nih.gov/pubmed/18535195 http://dx.doi.org/10.2337/dc08-0286 |
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