Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Multiplex Avidity Assay for Evaluating Thyroid Autoantibodies

Elena Savvateeva
* ORCID logo ,
Vera Sokolova
,
Marina Yukina
,
Nurana Nuralieva
ORCID logo ,
Elena Kulagina
,
Maria Isaeva
,
Ekaterina Troshina
,
Dmitry Gryadunov
Version 1 : Received: 9 November 2023 / Approved: 9 November 2023 / Online: 9 November 2023 (11:05:17 CET)

How to cite: Savvateeva, E.; Sokolova, V.; Yukina, M.; Nuralieva, N.; Kulagina, E.; Isaeva, M.; Troshina, E.; Gryadunov, D. Multiplex Avidity Assay for Evaluating Thyroid Autoantibodies. Preprints 2023, 2023110628. https://doi.org/10.20944/preprints202311.0628.v1 Savvateeva, E.; Sokolova, V.; Yukina, M.; Nuralieva, N.; Kulagina, E.; Isaeva, M.; Troshina, E.; Gryadunov, D. Multiplex Avidity Assay for Evaluating Thyroid Autoantibodies. Preprints 2023, 2023110628. https://doi.org/10.20944/preprints202311.0628.v1

Abstract

Autoantibodies (Abs) to thyroid antigens are associated with autoimmune thyroid disease (AITD), although sometimes they have no clear clinical significance. The aim of this study was to evaluate the avidity of thyroid Abs in the sera of AITD patients and non-AITD Abs carriers. A multiplex microarray-based assay was developed to detect the avidity of thyroid disease-associated Abs, including Abs to thyroperoxidase (TPO), thyroglobulin (Tg), pendrin, thyroid hormones T3 and T4, pyruvate kinase, and carbonic anhydrase 2. The method is based on indirect multiplex assay with an additional step involving the chaotrope destruction of the autoantibody-antigen complexes. The avidity index (AI) is then calculated. The method was tested on clinical cohort (n =117) and then confirmed by studies with another independent cohort (n =123). For the confirmation study, the median AI for AbTg was 29.9% among healthy Abs carriers, 52.6% for AITD patients, and 92.7% for non-AITD type 1 diabetes (T1D) patients. Median AI for AbTPO were 39.9% in healthy carriers, 73.4% in AITD patients, 83.2% in T1D patients, and 98.5% in AITD patients with thyroid neoplasm. The presence of high-avidity AbTg in T1D patients was also confirmed by a modified ELISA. Identification of high-avidity autoantibodies to thyroid proteins in non-thyroid T1D patients requires further studies.

Keywords

avidity; avidity index (AI); thyroid autoantibodies; protein microarray; multiplex assay; immunoassay

Subject

Biology and Life Sciences, Biochemistry and Molecular Biology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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