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CELL BASED ASSAY FOR MEASUREMENT OF BIOLOGICAL ACTIVITY OF ANTI TSHR AUTOIMMUNE ANTIBODIES

机译:基于细胞的抗TSHR自动免疫抗体生物活性测定

摘要

The invention relates to a bioassay for the determination of biological activity of stimulating autoimmune antibodies directed against the TSHR receptor comprising the steps of a) incubating TSHR expressing cells with a patient sample, b) detecting the intensity of a signal in the supernatant of the cells that is caused by a change of cAMP concentration in the cells resulting from TSHR stimulation, c) determining the stimulating activity of the anti TSHR autoimmune antibodies by comparing the signal intensity of step b) with the signal intensity obtained after incubation of a first reference sample with TSHR expressing cells; the invention also refers to a bioassay for the determination of the biological activity of blocking autoimmune antibodies directed against the thyroid stimulating hormone receptor (TSHR), comprising the steps of a) incubating TSHR expressing cells with a patient sample, b) detecting the intensity of a signal in the supernatant of the cells that is caused by a change of cAMP concentration in the cells resulting from TSHR stimulation, c) determining the blocking/stimulation activity of the anti TSHR auto antibodies by comparing the signal intensity of step b) with the signal intensities obtained after incubation of a first and a second reference sample with TSHR expressing cells, wherein the incubation sample of step a) and the second reference sample contain the same amount of a TSHR stimulating agent.
机译:本发明涉及用于测定刺激针对TSHR受体的自身免疫抗体的生物学活性的生物测定法,包括以下步骤:a)将TSHR表达细胞与患者样品一起温育,b)检测细胞上清液中信号的强度这是由TSHR刺激引起的细胞中cAMP浓度变化引起的,c)通过将步骤b)的信号强度与孵育第一个参考样品后获得的信号强度进行比较,确定抗TSHR自身免疫抗体的刺激活性TSHR表达细胞;本发明还涉及用于测定阻断针对甲状腺刺激激素受体(TSHR)的自身免疫抗体的生物学活性的生物测定法,其包括以下步骤:a)将TSHR表达细胞与患者样品一起温育,b)检测β-内酰胺酶的强度。由TSHR刺激引起的细胞上清液中cAMP浓度变化引起的细胞上清中的信号; c)通过将步骤b)的信号强度与步骤b)的信号强度进行比较,确定抗TSHR自身抗体的阻断/刺激活性在第一和第二参考样品与表达TSHR的细胞孵育后获得的信号强度,其中步骤a)的孵育样品和第二参考样品包含相同量的TSHR刺激剂。

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