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Detection and quantitation of host cell proteins in monoclonal antibody drug products using automated sample preparation and data-independent acquisition LC-MS/MS
Journal of Pharmaceutical Analysis ( IF 8.8 ) Pub Date : 2021-05-20 , DOI: 10.1016/j.jpha.2021.05.002
Lisa Strasser 1 , Giorgio Oliviero 1 , Craig Jakes 1, 2 , Izabela Zaborowska 1 , Patrick Floris 1 , Meire Ribeiro da Silva 1 , Florian Füssl 1 , Sara Carillo 1 , Jonathan Bones 1, 2
Affiliation  

Ensuring the removal of host cell proteins (HCPs) during downstream processing of recombinant proteins such as monoclonal antibodies (mAbs) remains a challenge. Since residual HCPs might affect product stability or safety, constant monitoring is required to demonstrate their removal to be below the regulatory accepted level of 100 ng/mg. The current standard analytical approach for this procedure is based on ELISA; however, this approach only measures the overall HCP content. Therefore, the use of orthogonal methods, such as liquid chromatography-mass spectrometry (LC-MS), has been established, as it facilitates the quantitation of total HCPs as well as the identification and quantitation of the individual HCPs present. In the present study, a workflow for HCP detection and quantitation using an automated magnetic bead-based sample preparation, in combination with a data-independent acquisition (DIA) LC-MS analysis, was established. Employing the same instrumental setup commonly used for peptide mapping analysis of mAbs allows for its quick and easy implementation into pre-existing workflows, avoiding the need for dedicated instrumentation or personnel. Thereby, quantitation of HCPs over a broad dynamic range was enabled to allow monitoring of problematic HCPs or to track changes upon altered bioprocessing conditions.



中文翻译:

使用自动化样品制备和数据独立采集 LC-MS/MS 检测和定量单克隆抗体药物产品中的宿主细胞蛋白

确保在单克隆抗体 (mAb) 等重组蛋白的下游加工过程中去除宿主细胞蛋白 (HCP) 仍然是一项挑战。由于残留的 HCP 可能会影响产品的稳定性或安全性,因此需要持续监测以证明其去除量低于监管认可的 100 ng/mg 水平。该程序的当前标准分析方法基于 ELISA;然而,这种方法仅测量整体 HCP 含量。因此,已经建立了正交方法的使用,例如液相色谱-质谱 (LC-MS),因为它有助于对总 HCP 进行定量以及对存在的单个 HCP 进行识别和定量。在本研究中,使用基于磁珠的自动样品制备进行 HCP 检测和定量的工作流程,结合数据独立采集(DIA)LC-MS分析,建立。使用通常用于 mAb 肽图分析的相同仪器设置,可以快速轻松地实施到预先存在的工作流程中,无需专用仪器或人员。因此,能够在广泛的动态范围内对 HCP 进行定量,以便监测有问题的 HCP 或跟踪改变生物工艺条件时的变化。

更新日期:2021-05-20
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