Lebert Dorothee, Picard Guillaume, Beau-Larvor Charlotte, Troncy Lysiane, Lacheny Christine, Maynadier Bernadette, Low Walter, Mouz Nicolas, Brun Virginie, Klinguer-Hamour Christine, Jaquinod Michel, Beck Alain
1PROMISE Advanced Proteomics, Grenoble, F-38040, France.
2Centre d'Immunologie Pierre Fabre (CIPF), 5 Av. Napoléon III, BP 60497, 74164 Saint-Julien-en-Genevois, France.
Bioanalysis. 2015;7(10):1237-51. doi: 10.4155/bio.15.56. Epub 2015 Apr 21.
In preclinical studies, monoclonal antibodies (mAbs) are traditionally assayed by ligand-binding-assays. Recently, quantitative liquid chromatography mass spectrometry (MS)-based assays have emerged which circumvent a number of challenges. These assays may also be multiplex, making them potentially compatible with pharmacokinetic assays for combined antibody therapies.
MATERIALS & METHODS: We combined a quantitative MS-based approach with the protein standard for absolute quantification (PSAQ™) strategy to simultaneously quantify three mAb variants presenting minor sequence differences. Stable isotopically labeled mAbs were produced and used as quantification standards. Titration curves were performed to assess the analytical performances of the method. LC-MS/MS and ELISA data were cross-compared.
The approach presented provides similar accuracy and precision than ELISA, while being multiplex and faster to develop. It has applications at all stages of the pharmaceutical development.
在临床前研究中，单克隆抗体（mAb）传统上通过配体结合测定法进行检测。最近，基于定量液相色谱质谱（MS）的检测方法出现了，它克服了许多挑战。这些检测方法也可以是多重的，使其有可能与联合抗体疗法的药代动力学检测兼容。
我们将基于定量MS的方法与用于绝对定量的蛋白质标准品（PSAQ™）策略相结合，以同时定量三种存在微小序列差异的mAb变体。制备了稳定同位素标记的mAb并用作定量标准品。进行滴定曲线以评估该方法的分析性能。对LC-MS/MS和ELISA数据进行交叉比较。
所提出的方法与ELISA具有相似的准确性和精密度，同时具有多重性且开发速度更快。它在药物开发的各个阶段都有应用。
