van Strien Jolinde, Dijk Lisanne, Atiqi Sadaf, Schouten Rogier, Bloem Karien, Wolbink Gerrit Jan, Loeff Floris, Rispens Theo
Department of Immunopathology, Sanquin Research, Amsterdam, the Netherlands; Landsteiner Laboratory, Academic Medical Centre, University of Amsterdam, the Netherlands.
Biologics Laboratory, Sanquin Diagnostic Services, Amsterdam, the Netherlands.
J Immunol Methods. 2023 Mar;514:113436. doi: 10.1016/j.jim.2023.113436. Epub 2023 Jan 28.
Accurate anti-drug antibody (ADA) measurements in patient sera requires dissociation of ADA-drug complexes combined with sensitive and specific ADA detection. Bridging type immunoassays are often used despite several disadvantages associated with this approach. A good drug-tolerant alternative is the acid-dissociation radioimmunoassay (ARIA), but this method is not easily implemented in most labs as specialized facilities are required for working with radioactive materials. We describe an innovative method for ADA detection that combines the advantages of antigen binding tests like the ARIA with the convenience of regular immunoassays. This acid-dissociation lanthanide-fluorescence immunoassay (ALFIA) involves dissociation of ADA-drug complexes, followed by binding to an europium-labeled drug derivative and subsequently an IgG pulldown on Sepharose beads. After europium elution, detection is achieved by measuring time-resolved fluorescence originating from europium chelate complexes. We measured anti-adalimumab ADA levels in sera of 94 rheumatoid arthritis patients using the ALFIA and showed this method to be highly drug tolerant, sensitive and specific for anti-adalimumab ADAs.
准确测量患者血清中的抗药物抗体（ADA）需要解离ADA-药物复合物，并结合灵敏且特异的ADA检测方法。尽管桥接型免疫测定法存在一些缺点，但仍经常被使用。一种良好的耐药物替代方法是酸解离放射免疫测定法（ARIA），但由于处理放射性物质需要专门的设施，这种方法在大多数实验室中不易实施。我们描述了一种创新的ADA检测方法，该方法结合了抗原结合试验（如ARIA）的优点和常规免疫测定法的便利性。这种酸解离镧系元素荧光免疫测定法（ALFIA）包括解离ADA-药物复合物，然后与铕标记的药物衍生物结合，随后在琼脂糖珠上进行IgG下拉。铕洗脱后，通过测量铕螯合物复合物产生的时间分辨荧光来实现检测。我们使用ALFIA测量了94例类风湿性关节炎患者血清中的抗阿达木单抗ADA水平，并表明该方法对抗阿达木单抗ADA具有高度的耐药物性、灵敏性和特异性。
