Development of a quantitative, high-throughput cell-based enzyme-linked immunosorbent assay for detection of colony-stimulating factor-1 receptor tyrosine kinase inhibitors

Nanomediator–Effector Cascade Systems for Amplified Protein Kinase Activity Imaging and Phosphorylation‐Induced Drug Release In Vivo

“…The ability to probe the spatiotemporal dynamics of kinase activity facilitates a better understanding of many facets of cellular functions in certain physiopathological conditions [6–8] . Antibody‐based assays including Western blot and enzyme‐linked immunosorbent assay (ELISA) were first invented to elucidate the quantity of kinase‐mediated phosphorylation, but these approaches lack temporal or spatial resolution at the cellular level [9, 10] . In the past two decades, masses of genetically encoded fluorescent reporters capable of continuously monitoring kinase activities in live cells and even in vivo were designed by fusing fluorescent proteins (FPs) or luciferase with substrate sequence and phosphoamino acid binding domain [11–14] .…”

“…[6][7][8] Antibody-based assays including Western blot and enzyme-linked immunosorbent assay (ELISA) were first invented to elucidate the quantity of kinasemediated phosphorylation, but these approaches lack temporal or spatial resolution at the cellular level. [9,10] In the past two decades,m asses of genetically encoded fluorescent reporters capable of continuously monitoring kinase activities in live cells and even in vivo were designed by fusing fluorescent proteins (FPs) or luciferase with substrate sequence and phosphoamino acid binding domain. [11][12][13][14] Further efforts were made on upgrading kinase activity reporters in the following years in terms of elevating the sensitivity and expanding the kinase tracking channels.…”

Nanomediator–Effector Cascade Systems for Amplified Protein Kinase Activity Imaging and Phosphorylation‐Induced Drug Release In Vivo

“…[6][7][8] Antibody-based assays including Western blot and enzyme-linked immunosorbent assay (ELISA) were first invented to elucidate the quantity of kinasemediated phosphorylation, but these approaches lack temporal or spatial resolution at the cellular level. [9,10] In the past two decades,m asses of genetically encoded fluorescent reporters capable of continuously monitoring kinase activities in live cells and even in vivo were designed by fusing fluorescent proteins (FPs) or luciferase with substrate sequence and phosphoamino acid binding domain. [11][12][13][14] Further efforts were made on upgrading kinase activity reporters in the following years in terms of elevating the sensitivity and expanding the kinase tracking channels.…”

Utilization of luminescent technology to develop a kinase assay: Cdk4 as a model system

“…Radioactive methods have been the most used approaches [1][2][3] so far. On the other hand, several non radioactive methods to evaluate kinase activity have been developed, based on fluorescent polarization technology [4] or on the use of specific anti-phospho-serine, -tyrosine or -threonine antibodies [5]. These techniques show important drawbacks: highly purified enzymes or specific anti-phospho antibodies are necessary, implying high costs and laborious technology, especially when analysing a kinase substrate with multiple phosphorylation sites.…”