Marin Clémence, Noé Gaëlle, Schlemmer Dimitri, Beaulieu Quentin, Robidou Pascal, Mansour Bochra, Hirtz Christophe, Vialaret Jérôme, Antignac Marie, Moyon Quentin, Benameur Neila, Amoura Zahir, Zahr Noël
AP-HP Sorbonne Université, Pitié-Salpêtrière Hospital, Department of Pharmacology, CIC-1901, Pharmacokinetics and Therapeutic Drug Monitoring Unit, UMR-S 1166, F-75013 Paris, France.
AP-HP. Sorbonne Université, Laboratoire de suivi thérapeutique pharmacologique spécialisé, F-75013 Paris, France.
J Pharm Biomed Anal. 2023 Nov 30;236:115730. doi: 10.1016/j.jpba.2023.115730. Epub 2023 Sep 17.
Belimumab is a monoclonal antibody against B cell activating factor (BLyS). This monoclonal antibody (mAb) has been shown to be effective in reducing disease activity in patients with systemic lupus erythematosus (SLE). Belimumab is available in two forms as a lyophilized powder for intravenous (IV) use, or single-dose syringe for subcutaneous (SC) use. The aim of this study was to develop and validate a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for quantitation of belimumab in human serum.
All analyses relied on nano-surface and molecular-orientation limited (nSMOL) proteolysis coupled with LC-MS/MS. Quantifications was performed in multiple reactions monitoring (MRM) mode, and electrospray ionization was conducted in positive mode.
Belimumab was quantified with signature peptide QAPGQGLEWMGGIPFGTAK and normalized using P14R. The total run time per assay was 10 min. Linearity was measured from 5 to 800 μg/mL (r² > 0.995). Accuracy and precision based on three quality control levels range from 11.2 - 9.51 % and 1.24 - 13.12 % respectively. The carryover was less than 7 %. In all, 87 patient samples were processed (65, IV; 22, SC). Mean concentration of belimumab was significantly higher for SC (93.0 ± 74.0 µg/mL) than for IV (67.4 ± 38.9 µg/mL) administration.
We have developed the first method of belimumab quantification combining LC-MS/MS and nSMOL proteolysis. It can be used for future clinical pharmacokinetic studies of belimumab and for investigating the relationship between belimumab concentration, efficacy, and toxicity in SLE patients.
贝利尤单抗是一种抗B细胞活化因子（BLyS）的单克隆抗体。这种单克隆抗体已被证明可有效降低系统性红斑狼疮（SLE）患者的疾病活动度。贝利尤单抗有两种剂型，一种是用于静脉注射（IV）的冻干粉末，另一种是用于皮下注射（SC）的单剂量注射器。本研究的目的是开发并验证一种液相色谱 - 串联质谱（LC-MS/MS）方法，用于定量测定人血清中的贝利尤单抗。
所有分析均依赖于纳米表面和分子取向受限（nSMOL）蛋白酶解结合LC-MS/MS。定量在多反应监测（MRM）模式下进行，电喷雾电离以正模式进行。
使用特征肽QAPGQGLEWMGGIPFGTAK对贝利尤单抗进行定量，并使用P14R进行归一化。每次测定的总运行时间为10分钟。线性范围为5至800μg/mL（r²>0.995）。基于三个质量控制水平的准确度和精密度分别为11.2 - 9.51%和1.24 - 13.12%。残留率小于7%。总共处理了87份患者样本（65份静脉注射样本；22份皮下注射样本）。皮下注射（SC）给药的贝利尤单抗平均浓度（93.0±74.0μg/mL）显著高于静脉注射（IV）给药（67.4±38.9μg/mL）。
我们开发了第一种结合LC-MS/MS和nSMOL蛋白酶解的贝利尤单抗定量方法。它可用于未来贝利尤单抗的临床药代动力学研究，以及研究SLE患者中贝利尤单抗浓度、疗效和毒性之间的关系。
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