Development of a general method for quantifying IgG-based therapeutic monoclonal antibodies in human plasma using protein G purification coupled with a two internal standard calibration strategy using LC-MS/MS
Development of a general method for quantifying IgG-based therapeutic monoclonal antibodies in human plasma using protein G purification coupled with a two internal standard calibration strategy using LC-MS/MS
复制标题
DOI：
10.1016/j.aca.2018.02.040
复制
发表时间：
2018-08-17
影响因子：
6.2
通讯作者：
Kuo, Ching-Hua
中科院分区：
文献类型：
--
作者：
Chiu, Huai-Hsuan;Liao, Hsiao-Wei;Kuo, Ching-Hua
近年来，单克隆抗体（mAb）药物在治疗各种疾病中引起了极大的兴趣。免疫球蛋白G（IgG）代表了已被美国食品药品监督管理局（FDA）批准的mAb药物的高百分比。为了便于治疗药物监测和药代动力学/药效学研究，我们开发了一种通用的液相色谱-串联质谱（LC-MS/MS）方法来定量人血浆中基于IgG的mAb的浓度。选择三种基于IgG的药物（贝伐单抗、纳武单抗和派姆单抗）来证明我们的方法。蛋白G珠用于样品预处理，因为它们具有捕获基于IgG的药物的通用能力。通过使用LC-MS/MS定量胰蛋白酶消化后获得的替代肽。为了校准LC-MS/MS分析期间发生的样品制备误差和基质效应，我们使用了两种内标（IS）方法，包括基于IgG的药物-IS托珠单抗和柱后输注IS。发现使用两种内标物有效地提高了定量准确度，对于在三种不同浓度下测试的所有mAb药物，定量准确度在15%以内。对该方法的精密度、准确度、线性和灵敏度进行了验证。该方法在临床样品中的成功应用证明了其在临床分析中的适用性。预计这种通用方法可以应用于其他基于mAb的药物，用于精确医学和临床研究。(c)2018爱思唯尔B. V.保留所有权利。
Monoclonal antibody (mAb) drugs have generated much interest in recent years for treating various diseases. Immunoglobulin G (IgG) represents a high percentage of mAb drugs that have been approved by the Food and Drug Administration (FDA). To facilitate therapeutic drug monitoring and pharmacokinetic/pharmacodynamic studies, we developed a general liquid chromatography-tandem mass spectrometry (LC-MS/MS) method to quantify the concentration of IgG-based mAbs in human plasma. Three IgG-based drugs (bevacizumab, nivolumab and pembrolizumab) were selected to demonstrate our method. Protein G beads were used for sample pretreatment due to their universal ability to trap IgG-based drugs. Surrogate peptides that were obtained after trypsin digestion were quantified by using LC-MS/MS. To calibrate sample preparation errors and matrix effects that occur during LC-MS/MS analysis, we used two internal standards (IS) method that include the IgG-based drug-IS tocilizumab and post-column infused IS. Using two internal standards was found to effectively improve quantification accuracy, which was within 15% for all mAb drugs that were tested at three different concentrations. This general method was validated in term of its precision, accuracy, linearity and sensitivity for 3 demonstration mAb drugs. The successful application of the method to clinical samples demonstrated its' applicability in clinical analysis. It is anticipated that this general method could be applied to other mAb-based drugs for use in precision medicine and clinical studies. (c) 2018 Elsevier B.V. All rights reserved.