BartsMS, Blizard Institute, Barts and the London School of Medicine and Dentistry, Queen Mary University of London, London, E1 2AT, United Kingdom.
School of Biological and Chemical Sciences, Queen Mary University of London, Mile End Road, London, E1 4NS, United Kingdom.
Sci Rep. 2020 Feb 5;10(1):1860. doi: 10.1038/s41598-020-58041-3.
The occurrence of anti-drug antibodies following administration of therapeutic monoclonal antibody to patients is a growing problem that is attracting attention from frontline clinicians. Ideally, an initial indicative point of care test would provide guidance to seek testing approved by the regulatory authorities. Here we describe a platform for the detection of IgG anti-drug antibodies that may provide an initial screen for all therapeutic monoclonal antibodies. Synthetic genes encoding Nanoluciferase polypeptides were inserted between the variable heavy and light domain encoding region of known antibody drugs (alemtuzumab and adalimumab) to generate recombinant single chain GloBodies, which retain the drug antibody paratopes and Nanoluciferase activity. In the presence of anti-drug antibodies, the GloBody is bound by specific IgG in the sample. These complexes are captured on immobilised Protein G and the luciferase activity determined. The amount of light generated being indicative of the anti-drug IgG antibody levels in serum. It should be possible to assemble GloBody reagents for all therapeutic monoclonal antibodies and adapt the capture phase to include additional specific isotypes. The assay has the potential to be developed for use with a drop of blood allowing initial pre-screening in a point of care setting.
在将治疗性单克隆抗体施用于患者后，抗药物抗体的出现是一个日益引起关注的问题，引起了一线临床医生的关注。理想情况下，初步的即时护理测试将为寻求监管机构批准的测试提供指导。在这里，我们描述了一种用于检测 IgG 抗药物抗体的平台，该平台可能为所有治疗性单克隆抗体提供初步筛选。在已知抗体药物（阿仑单抗和阿达木单抗）的可变重链和轻链编码区之间插入编码 Nanoluciferase 多肽的合成基因，以生成保留药物抗体表位和 Nanoluciferase 活性的重组单链 GloBody。在存在抗药物抗体的情况下，样品中的特异性 IgG 结合 GloBody。将这些复合物捕获在固定化的 Protein G 上，并测定荧光素酶活性。产生的光量指示血清中抗药物 IgG 抗体的水平。应该有可能为所有治疗性单克隆抗体组装 GloBody 试剂，并适应捕获阶段以包括其他特异性同种型。该测定法有可能开发用于一滴血，允许在即时护理环境中进行初步预筛选。
