Validating parameters of a luciferase reporter gene assay to measure neutralizing antibodies to IFNβ in multiple sclerosis patients
Validating parameters of a luciferase reporter gene assay to measure neutralizing antibodies to IFNβ in multiple sclerosis patients
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DOI：
10.1016/j.jim.2008.03.014
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发表时间：
2008-07-31
影响因子：
2.2
通讯作者：
Oger, Joel
中科院分区：
文献类型：
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作者：
Lam, Regina;Farrell, Rachel;Oger, Joel
在一些接受干扰素治疗的多发性硬化症（MS）患者中可能出现中和抗体（nab）。NAb降低药物生物利用度，高滴度降低药物疗效。我们描述了R. Farrell和G. Giovannoni荧光素酶报告基因测定法测定nab对INF β的有效性。我们用优化的荧光素酶方法检测了163例IFN β治疗的MS患者血清，并将结果与参考细胞病变效应（CPE）方法的结果进行了比较，该方法使用A549细胞和脑心肌炎病毒（EMCV）。结合抗体（BAbs）使用捕获ELISA作为CPE试验中nab的筛选试验来测量。荧光素酶法和ELISA/CPE法测定NAb状态无显著差异。荧光素酶法和A549/EMCV CPE法获得的Log10 NAb滴度相关性非常好。效价的检测间变异系数为17.8 ~ 29.3%，检测下变异系数为6.3 ~ 15.2%。荧光素酶测定是可靠的，适当的敏感性和需要更少的时间比目前可用的NAb方法。(C) 2008 Elsevier B.V.版权所有
Neutralizing antibodies (NAbs) can occur in some multiple sclerosis (MS) patients receiving interferon beta (IFN beta) therapy. NAbs reduce drug bioavailabity and high NAb titers reduce drug efficacy. We describe the validation of the R. Farrell and G. Giovannoni luciferase reporter gene assay to measure NAbs to INF beta. We assayed 163 sera from IFN beta treated MS patients with an optimized luciferase method and compared the results to those obtained with the reference cytopathic effect (CPE) method using A549 cells and an encephalomyocarditis virus (EMCV). Binding antibodies (BAbs) were measured using a capture ELISA as a screening test for NAbs in the CPE assay. NAb status measured by the luciferase and the ELISA/CPE method did not yield a significant difference. Log10 NAb titers obtained from the luciferase assay and the A549/EMCV CPE methods correlated very well. The inter-assay coefficient of variation for titers was between 17.8-29.3%, and the infra-assay coefficient of variation was between 6.3-15.2%. The luciferase assay is reliable, appropriately sensitive and requires less time than the currently available NAb methods. (C) 2008 Elsevier B.V. All rights reserved.