Improved Specificity of Glutamate Decarboxylase 65 Autoantibody Measurement Using Luciferase-Based Immunoprecipitation System Assays.

Autor:	Wyatt RC; Diabetes and Metabolism Unit, Translational Health Sciences, University of Bristol, Bristol, U.K., Grace SL; Diabetes and Metabolism Unit, Translational Health Sciences, University of Bristol, Bristol, U.K., Brigatti C; San Raffaele Diabetes Research Institute, IRCCS San Raffaele Scientific Institute, Milan, Italy., Marzinotto I; San Raffaele Diabetes Research Institute, IRCCS San Raffaele Scientific Institute, Milan, Italy., Gillard BT; Diabetes and Metabolism Unit, Translational Health Sciences, University of Bristol, Bristol, U.K., Shoemark DK; School of Biochemistry, University of Bristol, Bristol, U.K., Chandler K; Diabetes and Metabolism Unit, Translational Health Sciences, University of Bristol, Bristol, U.K., Achenbach P; Institute of Diabetes Research, Helmholtz Munich, German Center for Environmental Health, Munich, Germany.; Technical University of Munich, School of Medicine, Klinikum rechts der Isar, Forschergruppe Diabetes, Munich, Germany., Piemonti L; Division of Genetics and Cell Biology, IRCCS San Raffaele Scientific Institute, Milan, Italy., Long AE; Diabetes and Metabolism Unit, Translational Health Sciences, University of Bristol, Bristol, U.K., Gillespie KM; Diabetes and Metabolism Unit, Translational Health Sciences, University of Bristol, Bristol, U.K., Lampasona V; Division of Genetics and Cell Biology, IRCCS San Raffaele Scientific Institute, Milan, Italy., Williams AJK; Diabetes and Metabolism Unit, Translational Health Sciences, University of Bristol, Bristol, U.K.
Jazyk:	angličtina
Zdroj:	Diabetes [Diabetes] 2024 Apr 01; Vol. 73 (4), pp. 565-571.
DOI:	10.2337/db23-0550
Abstrakt:	Autoantibodies to glutamate decarboxylase (GADA) are widely used in the prediction and classification of type 1 diabetes. GADA radiobinding assays (RBAs) using N-terminally truncated antigens offer improved specificity, but radioisotopes limit the high-throughput potential for population screening. Luciferase-based immunoprecipitation system (LIPS) assays are sensitive and specific alternatives to RBAs with the potential to improve risk stratification. The performance of assays using the Nanoluc luciferase (Nluc)-conjugated GAD65 constructs, Nluc-GAD65(96-585) and full length Nluc-GAD65(1-585), were evaluated in 434 well-characterized serum samples from patients with recent-onset type 1 diabetes and first-degree relatives. Nonradioactive, high-throughput LIPS assays are quicker and require less serum than RBAs. Of 171 relatives previously tested single autoantibody positive for autoantibodies to full-length GAD65 by RBA but had not progressed to diabetes, fewer retested positive by LIPS using either truncated (n = 72) or full-length (n = 111) antigen. The Nluc-GAD65(96-585) truncation demonstrated the highest specificity in LIPS assays overall, but in contrast to RBA, N-terminus truncations did not result in a significant increase in disease-specificity compared with the full-length antigen. This suggests that binding of nonspecific antibodies is affected by the conformational changes resulting from addition of the Nluc antigen. Nluc-GAD65(96-585) LIPS assays offer low-blood-volume, high-specificity GADA tests for screening and diagnostics. (© 2024 by the American Diabetes Association.)
Databáze:	MEDLINE
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