Macroporous hydrogel micropillars for quantifying Met kinase activity in cancer cell lysates

摘要: Overactive and overexpressed kinases have been implicated in the cause progression of many cancers. Kinase inhibitors offer a targeted approach for treating cancers associated with increased or deregulated kinase activity. Often, however, cancer cells exhibit initial resistance to these evolve develop during treatment. Additionally, any one tissue type are typically heterogeneous their oncogenesis mechanisms, thus diagnosis particular does not necessarily provide insight into what therapies may be effective. For example, while some lung that overexpress epidermal growth factor receptor (EFGR) respond treatment EGFR inhibitors, overexpression hyperactivity Met correlates inhibitors. Here we describe microfluidic-based assay quantifying activity cell lysates eventual goals predicting responsiveness monitoring development In this assay, immobilized phosphorylation substrate macroporous hydrogel micropillars. We then exposed micropillars lysate detected using fluorescently conjugated antibody. This is able quantify whole from as few 150 cells. It can also detect expressing overactive background up 75% non-cancerous inhibition by Met-specific SU11274 PHA665752, suggesting predictive capability cellular response