A novel third-generation TSH receptor antibody (TRAb) enzyme-linked immunosorbent assay based on a murine monoclonal TSH receptor-binding antibody

Abstract

TSH receptor (TSHR) autoantibody (TRAb) is the serological hallmark of Graves' disease (GD). Third-generation enzyme-linked immunosorbent assays (ELISAs) using monoclonal TRAbs instead of TSH have been found useful for TRAb analysis recently. For the first time, a mouse monoclonal antibody (mAb) against TSHR was analyzed for TRAb detection and compared with human mAb M22 and TSH by the same competitive binding assay technique. A mouse monoclonal antibody (T7) binding to the TSH receptor and inhibiting TSH binding was generated and used for TRAb analysis in a third-generation ELISA. Obtained TRAb levels were compared with a second-generation TRAb assay employing bovine TSH and a third-generation assay with human mAb M22 as TSHR-binding reagents by investigating 89 patients with GD, 56 with Hashimoto's thyroiditis (HT), 73 with non-autoimmune thyroid diseases, 17 with rheumatoid arthritis, and 100 healthy subjects. The T7-based TRAb ELISA did not reveal a significantly different assay performance (area under the curve [AUC]) in contrast to the TSH and M22-based TRAb ELISAs by receiver operating characteristic (ROC) curve analysis (AUC-T7 0.967, AUC-TSH 0.972, AUC-M22 0.958, p > 0.05, respectively). After adjustment of cutoffs by ROC, all three TRAb ELISAs demonstrated sensitivities and specificities above 89.9% and 96.0%, respectively. Both thirdgeneration TRAb ELISAs showed a tendency for a higher prevalence of TRAb positives in HT in contrast to the secondgeneration ELISA. Mouse mAbs against the TSHR may be used for the reliable detection of TRAb by third-generation TRAb ELISA. The earlier reported higher sensitivity of third-generation TRAb ELISA in GD needs to be considered in the context of a slightly lower specificity regarding HT.

Relationship between the night shift work and thyroid disorders: A systematic review and meta-analysis

“…We included all the studies discussing the relationship between the night shift work and the thyroid disorders during the last ten years. Although the shift work is under investigation for many decades, we decided to restrict our analysis to a short timeframe since the 3rd generation assay, which has spread worldwide in the last 10 years, now ensures an excellent analytical sensitivity, able to determine very low TSH concentrations (Roggenbuck et al 2018). No restrictions were considered regarding age, sex, language, and ethnicity of the population.…”

The effect of radioiodine treatment on the characteristics of TRAb in Graves’ disease

“…Although a positive TRAb test result suggests the presence of TSAb or TBAb, it is reasonable to presume that a positive result in a patient with hyperthyroidism is due to TSAb. TSAb behaves like TSH and stimulates the synthesis of thyroid hormone by binding to TSHR, which leads to hyperthyroidism [ 8 ]. TSAb also causes diffuse, hypervascular goiter in many GD patients.…”

Third generation radioimmunoassay (RIA) for TSH receptor autoantibodies (TRAb) – one step less, similar results?

“…The mouse monoclonal TRAb T7 recognizing the TSH binding site of the TSHR was employed for the detection of TRAb in human sera by a novel 3 rd -generation RIA technique. Human TSHR was immobilized to polystyrene tubes by a mouse monoclonal TRAb not interfering with the TSH-binding site as reported elsewhere [20]. In contrast to the current two-step TRAb RIA procedure, 100 µL of neat patient serum or calibrators were simultaneously incubated with 50 µL of start buffer and 100 µL of 125 I-labelled T7 in the TSHR coated tubes at RT for 2 hours while shaking.…”

“…The human stimulating monoclonal TRAb M22 was the first to be used in an ELISA to detect TRAb [25,33]. It was later shown that other monoclonal TRAb, such as the murine T7, were not inferior to the former in similar assay designs [20].…”