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Kinase Inhibitor Screening In Self-assembled Human Protein Microarrays
Published on: October 23, 2019
Screening kinase inhibitors with a microarray-based fluorescent and resonance light scattering assay
Tao Li1, Dianjun Liu, Zhenxin Wang
1State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, 130022, China.
Analytical Chemistry
|March 6, 2010
Summary
This study introduces a novel microarray assay for screening kinase inhibitors using fluorescence and resonance light scattering (RLS). The assay effectively identifies kinase inhibitors and determines their binding strength and type.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Assay Development
Background:
- Kinase inhibitors are crucial in drug discovery.
- High-throughput screening (HTS) methods are needed for efficient inhibitor identification.
- Existing assays may lack sensitivity or require complex procedures.
Purpose of the Study:
- To develop a versatile microarray-based spectroscopic assay for screening kinase inhibitors.
- To utilize dual readout principles: fluorescence and resonance light scattering (RLS).
- To demonstrate the assay's utility in high-throughput screening and quantitative analysis.
Main Methods:
- Development of a microarray assay employing biotinylated antibodies for phosphorylation detection.
- Integration of gold nanoparticles and silver deposition for enhanced RLS signal.
- Utilization of avidin-conjugated fluorescein as a fluorescent probe.
- Application of standard avidin-biotin chemistry for antibody-nanoparticle conjugation.
Main Results:
- Successful development of assays for serine kinase (PKA) and tyrosine kinase (LCK).
- Demonstrated utility in high-throughput screening of a commercial kinase inhibitor library (80 compounds).
- Achieved satisfactory screening results with quantitative determination of inhibitor binding strength and type (Type I).
Conclusions:
- The developed microarray assay is effective for screening kinase inhibitors.
- The dual fluorescence and RLS readout offers versatility and sensitivity.
- The assay is suitable for high-throughput screening and detailed characterization of kinase inhibitors.
