Jourdil Jean-François, Némoz Benjamin, Gautier-Veyret Elodie, Romero Charlotte, Stanke-Labesque Françoise
University Hospital Grenoble-Alpes.
University Grenoble Alpes.
Ther Drug Monit. 2018 Aug;40(4):417-424. doi: 10.1097/FTD.0000000000000514.
Adalimumab (ADA) and infliximab (IFX) are therapeutic monoclonal antibodies targeting tumor necrosis factor-alpha (TNFα). They are used to treat inflammatory diseases. Clinical trials have suggested that therapeutic drug monitoring for ADA or IFX could improve treatment response and cost effectiveness. However, ADA and IFX were quantified by ELISA in all these studies, and the discrepancies between the results obtained raise questions about their reliability. We describe here the validation of a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous quantification of ADA and IFX in human samples.
Full-length antibodies labeled with stable isotopes were added to plasma samples as an internal standard. Samples were then prepared using Mass Spectrometry Immunoassay followed by trypsin digestion before ADA and IFX quantification by LC-MS/MS. ADA and IFX were quantified in serum from patients treated with ADA (n = 21) or IFX (n = 22), and the concentrations obtained were compared with those obtained with a commercial ELISA kit.
The chromatography run lasted 8.6 minutes, and the quantification range was 1-26 mg/L. The method was reproducible, repeatable, and accurate. For both levels of internal quality control, for ADA and IFX, interday and intraday coefficients of variation and accuracies were all within 15%, in accordance with FDA recommendations. No significant cross-contamination effect was noted. Good agreement was found between LC-MS/MS and ELISA results, for both ADA and IFX.
This LC-MS/MS method can be used for the quantification of ADA and IFX in a single analytical run and for the optimization of LC-MS/MS resource use in clinical pharmacology laboratories.
阿达木单抗（ADA）和英夫利昔单抗（IFX）是靶向肿瘤坏死因子-α（TNFα）的治疗性单克隆抗体。它们用于治疗炎症性疾病。临床试验表明，对ADA或IFX进行治疗药物监测可改善治疗反应并提高成本效益。然而，在所有这些研究中，ADA和IFX均通过酶联免疫吸附测定（ELISA）进行定量，所得结果之间的差异引发了对其可靠性的质疑。我们在此描述一种液相色谱-串联质谱（LC-MS/MS）方法用于同时定量人样本中ADA和IFX的验证。
将用稳定同位素标记的全长抗体作为内标添加到血浆样本中。然后使用质谱免疫测定法制备样本，接着进行胰蛋白酶消化，之后通过LC-MS/MS对ADA和IFX进行定量。对接受ADA治疗的患者（n = 21）或IFX治疗的患者（n = 22）的血清中的ADA和IFX进行定量，并将所得浓度与使用商用ELISA试剂盒获得的浓度进行比较。
色谱运行持续8.6分钟，定量范围为1-26 mg/L。该方法具有可重复性、可再现性和准确性。对于ADA和IFX的两个内部质量控制水平，日间和日内变异系数及准确度均在15%以内，符合美国食品药品监督管理局（FDA）的建议。未观察到明显的交叉污染效应。对于ADA和IFX，LC-MS/MS与ELISA结果之间均发现良好的一致性。
这种LC-MS/MS方法可用于在单次分析运行中对ADA和IFX进行定量，并可优化临床药理学实验室中LC-MS/MS资源的使用。
