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在Madin-Darby犬细胞蛋白质组中实验室间的变异性

Matthew D Harwood¹, Katharina Zettl², Manuel Weinheimer³

¹Certara UK Ltd (Simcyp Division), Sheffield S1 2BJ, U.K.

Molecular pharmaceutics

|June 7, 2023

概括

这项研究量化了跨实验室的Madin-Darby犬 (MDCK) 细胞中的蛋白质,揭示了上皮细胞模型中的变异性. 这些发现提供了一个数据库,以改善药物透性评估和体外-体外抽取 (IVIVE).

科学领域:

细胞生物学细胞生物学
蛋白质组学是指蛋白质组学.
药理学药理学是指药理学的学科.

背景情况:

马丁-达比犬 (MDCK) 细胞对于研究表皮功能和药物运输至关重要.
实验室之间MDCK细胞表型的变化会影响药物透性评估和体外-体外抽取 (IVIVE).

研究的目的:

来自多个实验室的MDCK细胞单层的全面蛋白质量定量.
建立一个数据库,以了解MDCK细胞模型中的实验室间的变异性.
通过表征控制单层来实现更准确的IVIVE.

主要方法:

在8个不同实验室的11个过成长的MDCK单层上使用总蛋白方法 (TPA) 进行蛋白质量定量.
分析了超过8000个蛋白质拷贝数和度.
估计关键的形态参数,如细胞性和体积.

主要成果:

在MDCK细胞表型中发现了实验室间的显著变异性.
关键药物载体的量化表达水平,其中SLC16A1 (MCT1) 和ABCC4 (MRP4) 是最丰富的.
证实了克劳丁-2水平与影响跨表皮抵抗的紧接节调节之间的联系.
由于低酶表达,MDCK细胞对异生菌的代谢责任性有限.

结论:

生成的蛋白质基因数据库为标准化MDCK细胞模型提供了宝贵的资源.
了解MDCK细胞变异性对于可靠的药物透性研究和IVIVE至关重要.
这些数据支持使用MDCK细胞作为模型系统,其清晰的表征对于可复制性至关重要.

关键词:

在ADME蛋白质的丰富性中,ADME蛋白质的丰富性没有标签的蛋白质组学.这就是MDCKK的MDCK.皮质蛋白质组是表皮细胞蛋白质组.总蛋白质分析 (TPA)

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