Iwamoto Noriko, Yokoyama Kotoko, Takanashi Megumi, Yonezawa Atsushi, Matsubara Kazuo, Shimada Takashi
Leading Technology of Bioanalysis and Protein Chemistry, Shimadzu Corporation, Kyoto, Japan.
Department of Clinical Pharmacology and Therapeutics, Kyoto University Hospital, Kyoto, Japan.
Curr Pharm Biotechnol. 2018;19(6):495-505. doi: 10.2174/1389201019666180703093517.
Infliximab (IFX) is a chimeric therapeutic monoclonal antibody targeting tumor necrosis factor alpha (TNFα)-mediated inflammatory immune diseases. However, despite of an initial good clinical response, decrease in response to long-term treatment is a common observation.
Recent studies suggest that IFX level in circulation has a correlation with clinical bioavailability. Therefore, the management of IFX dosage for individual manifestation by IFX monitoring may be valuable for the improvement of therapeutic response and outcomes.
In order to develop a broad IFX therapeutic monitoring in human serum, we have developed the validated IFX bioanalysis for RemicadeTM and its biosimilar product using our nano-surface and molecular-orientation limited proteolysis (nSMOL) technology coupled with liquid chromatographytandem mass spectrometry (LC-MS/MS). The nSMOL chemistry has a unique property of Fabselective proteolysis, and makes it possible a global bioanalysis for many monoclonal antibodies.
The quantitation range of IFX in serum was from 0.293 to 300 μg/ml with good linearity. Quantitation verification at the concentrations of 0.293, 0.879, 14.1 and 240 µg/ml was within 1.56- 7.53% of precision and 98.9-111% of accuracy using H-chain signature peptide SINSATHYAESVK. Moreover, cross-verified bioanalysis of Remicade quantitation using biosimilar standard, and its opposite combination, obtained an identical and inter-comparative results.
The nSMOL strategy has the potential as a practical therapeutic monitoring technology in IFX therapeutic applications.
英夫利昔单抗（IFX）是一种嵌合治疗性单克隆抗体，用于治疗肿瘤坏死因子α（TNFα）介导的炎性免疫疾病。然而，尽管初始临床反应良好，但长期治疗反应下降是常见现象。
近期研究表明，循环中IFX水平与临床生物利用度相关。因此，通过监测IFX来管理个体表现的IFX剂量，可能对改善治疗反应和结果具有重要价值。
为了在人血清中开展广泛的IFX治疗监测，我们利用纳米表面和分子取向受限蛋白水解（nSMOL）技术结合液相色谱-串联质谱（LC-MS/MS），开发了用于类克（RemicadeTM）及其生物类似药产品的经过验证的IFX生物分析方法。nSMOL化学具有Fab选择性蛋白水解的独特特性，使得对许多单克隆抗体进行全面生物分析成为可能。
血清中IFX的定量范围为0.293至300μg/ml，线性良好。使用重链特征肽SINSATHYAESVK，在0.293、0.879、14.1和240μg/ml浓度下进行定量验证，精密度在1.56%-7.53%之间，准确度在98.9%-111%之间。此外，使用生物类似药标准品对类克定量进行交叉验证生物分析，以及相反的组合，均获得了相同且可相互比较的结果。
nSMOL策略在IFX治疗应用中具有作为实用治疗监测技术的潜力。
