Multiplexed monitoring of therapeutic antibodies for inflammatory diseases using Fab-selective proteolysis nSMOL coupled with LC-MS
Multiplexed monitoring of therapeutic antibodies for inflammatory diseases using Fab-selective proteolysis nSMOL coupled with LC-MS
复制标题
DOI：
10.1016/j.jim.2019.06.014
复制
发表时间：
2019-09-01
影响因子：
2.2
通讯作者：
Shimada, Takashi
中科院分区：
文献类型：
--
作者：
Iwamoto, Noriko;Takanashi, Megumi;Shimada, Takashi
单克隆抗体加速了许多疾病治疗方案的出现，其中的分子机制已被详细阐明。因此，一种能够通用分析抗体以进行临床药代动力学和横断面研究的分析方法是必不可少的。我们开发了一种具有 Fab 选择性胰蛋白酶反应的通用抗体生物分析方法，称为纳米表面和分子定向有限（nSMOL）蛋白水解，可收集生物样品中的特定抗体特征肽。使用nSMOL方法，我们充分验证了许多抗体、Fc融合蛋白及其生物仿制药的生物分析。由于观察到缓解和复发，炎症免疫疾病通常需要长期的临床治疗。体循环中准确的抗体监测有助于改善临床结果。由于可以选择多种生物药品作为实际治疗方案，因此同时定量多种抗体的测定开发将适用于许多治疗监测。在本研究中，我们使用 nSMOL 正常反应条件和使用酸化还原加速条件的两种混合抗体（托珠单抗和美泊利单抗），如我们之前的论文中报道的。此外，这种多重检测已使用临床患者样本进行了验证。 nSMOL 方法能够同时量化人血清中的多种免疫抑制抗体，并且 nSMOL 可能适用于多种炎症免疫疾病的药物相互作用测定或治疗性抗体监测，以优化给药。
Monoclonal antibodies have accelerated the availability of treatment options for many diseases in which the molecular mechanism has been elucidated in detail. Therefore, an assay that can universally analyze antibodies for clinical pharmacokinetics and cross-sectional studies would be indispensable.We have developed a universal antibody bioanalysis with a Fab-selective tryptic reaction, named nano-surface and molecular-orientation limited (nSMOL) proteolysis, that collects the specific antibody signature peptides in biological samples. Using the nSMOL method, we have fully validated the bioanalysis of many antibodies, Fc-fusion proteins, and their biosimilars.Inflammatory immune diseases often require long-term clinical management because of the remission and relapse observed. Accurate antibody monitoring in systemic circulation could contribute to the improvement of clinical outcomes. Because several biopharmaceuticals can be selected as practical treatment options, the assay development that quantitates many antibodies simultaneously would be applicable in many theraprutic monitoring.In this study, we have validated the LC-MS bioanalysis method for seven-mixed antibodies (Infliximab, Adalimumab, Ustekinumab, Golimumab, Eculizumab, Etanercept, and Abatacept) using the nSMOL normal reaction condition and two-mixed antibodies (Tocilizumab and Mepolizumab) using the acidified reduction acceleration condition, as reported in our previous papers. Moreover, this multiplexed assay has been verified using clinical patient samples. The nSMOL approach enables the quantization of several immunosuppressive antibodies simultaneously in human serum, and nSMOL can potentially be applicable to the drug-drug interaction assays or therapeutic antibody monitoring of several inflammatory immune diseases to optimize administration.