Wallukat Gerd, Wenzel Katrin, Schimke Ingolf
Berlin Cures GmbH, Berlin, Germany.
Methods Mol Biol. 2019;1955:247-261. doi: 10.1007/978-1-4939-9148-8_19.
Autoantibodies directed against G-protein-coupled receptors (GPCR-AAB), an autoantibody type discovered in the 1970s, affect functionally their targets and are therefore called functional autoantibodies. GPCR-AAB are increasingly accepted as the origin or amplifier of various diseases. Here, we describe the present "gold standard" for measurement of GPCR-AAB in human blood. This bioassay monitors the chronotropic activity of GPCR-AAB by recording the spontaneous beating of cultured neonatal rat cardiomyocytes. The construction of this bioassay and its procedure and standardization for GPCR-AAB measurement are described in detail and also include the application of the bioassay for GPCR-AAB differentiation related to first the targeted receptors and IgG subclasses carrying the GPCR-AAB and second the extracellular receptor-binding site and specific epitopes targeted by the GPCR-AAB.
针对G蛋白偶联受体的自身抗体(GPCR-AAB)是20世纪70年代发现的一种自身抗体类型,它会在功能上影响其靶标,因此被称为功能性自身抗体。GPCR-AAB越来越被认为是各种疾病的起源或放大器。在这里,我们描述了目前用于测量人血液中GPCR-AAB的“金标准”。这种生物测定法通过记录培养的新生大鼠心肌细胞的自发搏动来监测GPCR-AAB的变时活性。详细描述了这种生物测定法的构建及其用于GPCR-AAB测量的程序和标准化,还包括该生物测定法在GPCR-AAB鉴别中的应用,首先是与携带GPCR-AAB的靶向受体和IgG亚类有关,其次是与GPCR-AAB靶向的细胞外受体结合位点和特定表位有关。