一种经验证的 LC-MS/MS 方法，用于在不进行 IgG 纯化和还原的情况下，通过快速胰蛋白酶消化定量测定大鼠血清中的达妥木单抗。
A validated LC-MS/MS method for the quantitation of daratumumab in rat serum using rapid tryptic digestion without IgG purification and reduction.
机构信息
Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201210, China; University of Chinese Academy of Sciences, Beijing 100049, China.
Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 201210, China; School of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, China.
出版信息
J Pharm Biomed Anal. 2024 Jun 15;243:116083. doi: 10.1016/j.jpba.2024.116083. Epub 2024 Mar 2.
Daratumumab, a humanized monoclonal antibody utilized in treating immunoglobulin light-chain amyloidosis and relapsed/refractory multiple myeloma, was quantified in rat serum through a simple, economical and effective liquid chromatography tandem-mass spectrometry (LC-MS/MS) method. A surrogate peptide, LLIYDASNR, derived from trypsin hydrolysis, was quantitatively analyzed with LLIYDASN [C, N] RAT as an internal standard. This corrected variations from sample pretreatment and mass spectrometry response, involving denaturation and trypsin hydrolysis in a two-step process lasting approximately 1 hour. Methodological validation demonstrated a linear range of 1 µg/mL to 1000 µg/mL in rat serum. Precision, accuracy, matrix effect, sensitivity, stability, selectivity, carryover, and interference met acceptance criteria. The validated LC-MS/MS approach was successfully applied to a pharmacokinetic study of daratumumab in rats at an intravenous dose of 15 mg/kg.
达雷妥尤单抗是一种人源化单克隆抗体，用于治疗免疫球蛋白轻链淀粉样变性和复发/难治性多发性骨髓瘤。通过一种简单、经济和有效的液相色谱串联质谱（LC-MS/MS）方法，对大鼠血清中的达雷妥尤单抗进行定量分析。通过内标 LLIYDASN [C, N] RAT，对来源于胰蛋白酶水解的替代肽 LLIYDASNR 进行定量分析。该方法校正了样品预处理和质谱响应过程中的变异，涉及大约 1 小时的两步变性和胰蛋白酶水解过程。方法学验证表明，大鼠血清中的线性范围为 1 µg/mL 至 1000 µg/mL。精密度、准确度、基质效应、灵敏度、稳定性、选择性、残留和干扰均符合可接受标准。该经验证的 LC-MS/MS 方法成功应用于大鼠静脉注射 15mg/kg 达雷妥尤单抗的药代动力学研究。
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