Bloem Karien, van Leeuwen Astrid, Verbeek Gerrit, Nurmohamed Michael T, Wolbink Gerrit Jan, van der Kleij Desiree, Rispens Theo
Sanquin Research, Amsterdam, The Netherlands; Landsteiner Laboratory, Academic Medical Centre, University of Amsterdam, The Netherlands; Sanquin Diagnostic Services, Amsterdam, The Netherlands.
Sanquin Diagnostic Services, Amsterdam, The Netherlands.
J Immunol Methods. 2015 Mar;418:29-38. doi: 10.1016/j.jim.2015.01.007. Epub 2015 Jan 27.
Drug interference complicates assessment of immunogenicity of biologicals and results in an underestimation of anti-drug antibody (ADA) formation. Drug-tolerant assays have the potential to overcome such limitations. However, to which extent drug-tolerant assays provide an unbiased picture of the antibody response to a biological is unknown. In this study, we compared the measurement of ADA to adalimumab in 94 consecutive adalimumab-treated rheumatoid arthritis patients using the traditional antigen binding test (ABT) and four different drug-tolerant assays, the Ph-shift anti-Idiotype Antigen binding test (PIA) and three newly developed assays for this study: an acid-dissociation radioimmunoassay (ARIA), a temperature-shift radioimmunoassay (TRIA) and an electrochemoluminescence-based assay (ECL). Our results indicate that drug-tolerant assays provide a fairly consistent view on the antibody formation: quantitatively, the results from all four assays correlate well (Spearman r > 0.9). However, the percentage of ADA-positive patients ranges from 51 to 66% between assays, with the ARIA identifying the highest number of patients as positive. These differences are largely due to patients making low amounts of ADA; if ADA levels were above ca. 100 AU/ml, a patient was identified as positive in all four assays. Adalimumab concentrations were significantly lower in ADA-positive samples. Taken together, the results indicate that these different drug-tolerant assays provide a similar and reasonably consistent view on ADA responses, which however, breaks down at the lower end of the detectable range, and highlight that ADA is best reported quantitatively. Furthermore, if an even more sensitive drug-tolerant assay could be developed, one would probably find additional positive samples that will predominantly contain very low levels of ADA.
药物干扰会使生物制品免疫原性的评估变得复杂，并导致对抗药物抗体（ADA）形成的低估。药物耐受检测有潜力克服此类限制。然而，药物耐受检测在多大程度上能提供对生物制品抗体反应的无偏倚情况尚不清楚。在本研究中，我们使用传统抗原结合试验（ABT）和四种不同的药物耐受检测方法，即Ph-位移抗独特型抗原结合试验（PIA）以及为本研究新开发的三种检测方法：酸解离放射免疫测定法（ARIA）、温度位移放射免疫测定法（TRIA）和基于电化学发光的测定法（ECL），对94例连续接受阿达木单抗治疗的类风湿性关节炎患者中ADA与阿达木单抗的检测进行了比较。我们的结果表明，药物耐受检测对抗体形成提供了相当一致的观点：在定量方面，所有四种检测方法的结果相关性良好（Spearman相关系数r>0.9）。然而，不同检测方法之间ADA阳性患者的百分比在51%至66%之间变化，其中ARIA检测出的阳性患者数量最多。这些差异很大程度上是由于产生低水平ADA的患者所致；如果ADA水平高于约100 AU/ml，所有四种检测方法都会将该患者判定为阳性。ADA阳性样本中的阿达木单抗浓度显著更低。总体而言，结果表明这些不同的药物耐受检测方法对ADA反应提供了相似且合理一致的观点，不过，在可检测范围的下限处这种一致性会被打破，并突出了ADA最好进行定量报告。此外，如果能够开发出更灵敏的药物耐受检测方法，可能会发现更多主要含有极低水平ADA的阳性样本。
