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Updated: May 4, 2026
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Probing High-density Functional Protein Microarrays to Detect Protein-protein Interactions
Published on: August 2, 2015
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Simultaneous measurement of 10,000 protein-ligand affinity constants using microarray-based kinetic constant assays
James P Landry1, Yiyan Fei, Xiangdong Zhu
1Department of Physics, University of California at Davis, Davis, CA 95616, USA.
Assay and Drug Development Technologies
|December 24, 2011
Summary
This study introduces a novel kinetic assay platform for microarrays, enabling high-throughput screening of molecular interactions. It moves beyond endpoint detection to measure binding kinetics, offering a more reliable method for identifying novel ligands.
Area of Science:
- Biochemistry
- Molecular Biology
- Analytical Chemistry
Background:
- Fluorescence-based microarray detection is useful for high-throughput screening but lacks kinetic information.
- Endpoint data reliability for binding affinity is limited by reaction conditions and processing.
Purpose of the Study:
- To develop a microarray assay platform capable of simultaneous measurement of binding curves for thousands of molecular targets.
- To transition microarray technology from endpoint detection to kinetic constant assays for improved biomolecular interaction analysis.
Main Methods:
- Utilized an ellipsometry-based optical scanner for label-free, simultaneous measurement of protein probe binding curves against 10,000 microarray targets.
- Extracted kinetic rate constants (k(on), k(off), k(a)) from binding curves to characterize probe-target interactions.
Main Results:
- Successfully measured binding curves for a protein probe against 10,000 molecular targets on a microarray.
- Characterized probe-target interactions using kinetic rate constants, providing more detailed information than endpoint detection.
- Achieved throughput comparable to national screening centers, demonstrating practical utility.
Conclusions:
- This work establishes a kinetic constant assay platform for microarrays, advancing beyond traditional endpoint detection.
- The platform enables practical, high-throughput screening of compound libraries for novel ligands and system-wide affinity profiling.
