Journal
ANALYTICAL BIOCHEMISTRY
Volume 422, Issue 2, Pages 79-88Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.ab.2011.12.048
Keywords
Biomarker; ECPKA; PKA; Homogeneous biomarker test; Protein kinase analysis; Time-gated luminescence
Funding
- Estonian Science Foundation [8230, 8055]
- Estonian Ministry of Education and Research [SF0180121s08, SF0180161s08]
- Kinasera OY
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A novel nonmetal optical probe ARC-1063 whose long-lifetime luminescence is induced by association with the target protein kinase is used for the measurement of the concentration of catalytic subunit of protein kinase A (PKAc) in complicated biological solutions. High affinity (K-D = 10 pM toward PKAc) and unique optical properties of the probe enable its application for the measurement of picomolar concentrations of PKAc in the presence of high concentrations of other proteins. The described assay is applicable in the high-throughput format with the instrument setups designed for lanthanide-based time-gated (time-resolved) luminescence methods. The assay is used for demonstration that extracellular PKAc (ECPKA) is present in plasma samples of all healthy persons and cancer patients but great care must be taken for procedures of treatment of blood samples to avoid disruption, damage, or activation of platelets in the course of plasma (or serum) preparation and conservation. (C) 2012 Elsevier Inc. All rights reserved.
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