Multicolored, Tb3+-Based Antibody-Free Detection of Multiple Tyrosine Kinase Activities

Andrew M. Lipchik, Minervo Perez, Wei Cui, Laurie L. Parker

Research output: Contribution to journalArticlepeer-review

8 Scopus citations

Abstract

Kinase signaling is a major mechanism driving many cancers. While many inhibitors have been developed and are employed in the clinic, resistance due to crosstalk and pathway reprogramming is an emerging problem. High-throughput assays to detect multiple pathway kinases simultaneously could better model these complex relationships and enable drug development to combat this type of resistance. We developed a strategy to take advantage of time-resolved luminescence of Tb3+-chelated phosphotyrosine-containing peptides, which facilitated efficient energy transfer to small molecule fluorophores conjugated to the peptides to produce orthogonally colored biosensors for two different kinases. This enabled multiplexed detection with high signal-to-noise in a high-throughput-compatible format. This proof-of-concept study provides a platform that could be applied to other lanthanide metal and fluorophore combinations to achieve even greater multiplexing without the need for phosphospecific antibodies.

Original languageEnglish (US)
Pages (from-to)7555-7558
Number of pages4
JournalAnalytical Chemistry
Volume87
Issue number15
DOIs
StatePublished - Aug 4 2015

Bibliographical note

Publisher Copyright:
© 2015 American Chemical Society.

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