In-depth analysis of monoclonal antibodies using microfluidic capillary electrophoresis and native mass spectrometry.,Journal of Pharmaceutical and Biomedical Analysis

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In-depth analysis of monoclonal antibodies using microfluidic capillary electrophoresis and native mass spectrometry.
Journal of Pharmaceutical and Biomedical Analysis ( IF 3.1 ) Pub Date : 2020-02-29 , DOI: 10.1016/j.jpba.2020.113218
Sara Carillo ₁ , Craig Jakes ₂ , Jonathan Bones ₂

Affiliation

Charge variant profiling of therapeutic proteins is required by the International Council for Harmonisation guidelines and is traditionally performed by capillary electrophoresis or ion exchange chromatography. Recently, improvements in the hyphenation of capillary electrophoresis with mass spectrometry and the introduction of mass spectrometry compatible background electrolytes has allowed the implementation of native mass spectrometric determination of the charge variant profile obtained from the electrophoretic separation. The low flow operation of the microfluidic electrophoretic platform significantly boosts mass spectrometric sensitivity and increases the dynamic range, even when using sample amounts as low as 1 ng in capillary. In the current study, rituximab, trastuzumab and bevacizumab drug products were analysed using the ZipChip microfluidic CE-ESI-MS platform that facilitated confident identification of proteoforms with an average mass accuracy of <15 ppm. Up to 52 proteoforms were identified for trastuzumab drug product, while rituximab sample revealed the presence of fragments and sialylated N-glycans. Overall, the CE-ESI-MS platform proved to be a fast and robust tool for therapeutic protein charge variant profiling and facilitated efficient coupling with native mass spectrometry for the generation of highly informative characterisation data.

中文翻译：

使用微流控毛细管电泳和天然质谱对单克隆抗体进行深入分析。

国际协调委员会指南要求对治疗性蛋白质进行电荷变异分析，传统上是通过毛细管电泳或离子交换色谱法进行的。近来，在毛细管电泳的质谱联用方面的改进以及与质谱兼容的背景电解质的引入，使得可以对通过电泳分离获得的电荷变化曲线进行天然质谱测定。即使在毛细管中使用低至1 ng的样品量时，微流体电泳平台的低流量操作也可显着提高质谱灵敏度并增加动态范围。在目前的研究中，利妥昔单抗使用ZipChip微流体CE-ESI-MS平台对曲妥珠单抗和贝伐单抗药物产品进行了分析，该平台可帮助可靠地鉴定平均质量准确度低于15 ppm的蛋白形式。曲妥珠单抗药物产品鉴定出多达52种蛋白形式，而利妥昔单抗样品显示存在片段和唾液酸化的N-聚糖。总体而言，CE-ESI-MS平台被证明是用于治疗性蛋白质电荷变异图谱的快速且强大的工具，并促进了与天然质谱的高效耦合，以生成高信息量的表征数据。而利妥昔单抗样品显示存在片段和唾液酸化的N-聚糖。总体而言，CE-ESI-MS平台被证明是用于治疗性蛋白质电荷变异图谱的快速且强大的工具，并促进了与天然质谱的高效耦合，以生成高信息量的表征数据。而利妥昔单抗样品显示存在片段和唾液酸化的N-聚糖。总体而言，CE-ESI-MS平台被证明是用于治疗性蛋白质电荷变异图谱的快速且强大的工具，并促进了与天然质谱的高效耦合，以生成高信息量的表征数据。

更新日期：2020-03-02

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