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Probing High-density Functional Protein Microarrays to Detect Protein-protein Interactions
Published on: August 2, 2015
Probing protein binding spectra with Fourier microfluidics.
C H Mastrangelo1, L D Williams, T Ghosh
1Electrical Engineering and Bioengineering Departments, University of Utah, Salt Lake City, UT 84112, USA. carlos.mastrangelo@utah.edu
Summary
This study introduces a microfluidic chip for Fourier transform analysis of biochemical interactions. The technology enables precise measurement of binding kinetics, offering a new tool for chemical analysis.
Area of Science:
- Biochemistry
- Microfluidics
- Analytical Chemistry
Background:
- Microfluidic chip technology advancements allow for the development of chemical spectrum analyzers.
- These analyzers can investigate binding interactions between various chemical entities.
- Fourier transform measurements offer a powerful method for analyzing dynamic biochemical processes.
Purpose of the Study:
- To implement a microfluidic chip specifically designed for Fourier transform measurements of biochemical interactions.
- To demonstrate the capability of this chip in probing binding kinetics.
- To validate the technique using a model biochemical system.
Main Methods:
- Development of a microfluidic chip integrating a chemical signal generator, flow cell, and binding sensor surface.
- Generation of periodic protein plugs in solution flowing through the cell to induce cycles of association and dissociation.
- Optical measurement of sensor activity using surface plasmon resonance (SPR) imaging to capture the phasor response.
Main Results:
- The microfluidic chip successfully generated periodic association and dissociation cycles of proteins to a functionalized gold surface.
- Surface plasmon resonance (SPR) imaging provided optical measurements of the sensor's phasor response.
- Analysis of the transfer function revealed a dominant pole at 10.2 mHz, yielding association and dissociation constants for carbonic anhydrase-II and its ligand.
Conclusions:
- The implemented microfluidic chip is suitable for Fourier transform measurements of biochemical interactions.
- The technique allows for the determination of binding kinetics, specifically association and dissociation constants.
- This technology offers a novel approach for analyzing chemical and biochemical binding events with high precision.
