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Integrated microfluidic and imaging platform for a kinase activity radioassay to analyze minute patient cancer
Cong Fang1, Yanju Wang, Nam T Vu
1Department of Molecular and Medical Pharmacology, Crump Institute for Molecular Imaging, University of California, Los Angeles, California 90095, USA.
Abstract:
Oncogenic kinase activity and the resulting aberrant growth and survival signaling are a common driving force of cancer. Accordingly, many successful molecularly targeted anticancer therapeutics are directed at inhibiting kinase activity. To assess kinase activity in minute patient samples, we have developed an immunocapture-based in vitro kinase assay on an integrated polydimethylsiloxane microfluidics platform that can reproducibly measure kinase activity from as few as 3,000 cells. For this platform, we adopted the standard radiometric (32)P-ATP-labeled phosphate transfer assay. Implementation on a microfluidic device required us to develop methods for repeated trapping and mixing of solid-phase affinity microbeads. We also developed a solid-state beta-particle camera imbedded directly below the microfluidic device for real-time quantitative detection of the signal from this and other microfluidic radiobioassays. We show that the resulting integrated device can measure ABL kinase activity from BCR-ABL-positive leukemia patient samples. The low sample input requirement of the device creates new potential for direct kinase activity experimentation and diagnostics on patient blood, bone marrow, and needle biopsy samples.
Insights
Researchers developed a microfluidic assay to measure cancer kinase activity in small patient samples. This innovation enables real-time diagnostics using minimal cellular material for targeted cancer therapies.
Area of Science:
- Biochemistry
- Molecular Biology
- Oncology
Background:
- Aberrant kinase activity drives cancer growth and survival.
- Targeting kinase activity is a key strategy in molecular cancer therapy.
- Assessing kinase activity in small patient samples is challenging.
Purpose of the Study:
- To develop a sensitive assay for measuring kinase activity in minute patient samples.
- To implement this assay on an integrated microfluidics platform.
- To enable real-time kinase activity diagnostics.
Main Methods:
- Developed an immunocapture-based in vitro kinase assay on a polydimethylsiloxane microfluidics platform.
- Utilized a radiometric (32)P-ATP-labeled phosphate transfer assay.
- Integrated a solid-state beta-particle camera for real-time detection.
Main Results:
- The microfluidics platform reproducibly measured kinase activity from as few as 3,000 cells.
- Successfully measured ABL kinase activity in BCR-ABL-positive leukemia patient samples.
- Demonstrated the potential for direct kinase activity experimentation on patient samples.
Conclusions:
- The developed microfluidic device enables sensitive kinase activity measurement from minimal patient samples.
- This technology facilitates direct kinase activity diagnostics on blood, bone marrow, and needle biopsies.
- Offers new potential for personalized medicine and targeted cancer therapies.
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