Abstract: PUB090
Optimizing Microfluidic Gradient Generating Surface Plasmon Resonance (SPR) Chips for High-Throughput Biochemical Reaction Kinetics in Renal Physiology
Session Information
Category: Bioengineering
- 400 Bioengineering
Authors
- Molnar, Daniela, New York Institute of Technology College of Osteopathic Medicine at Arkansas State University, Jonesboro, Arkansas, United States
- Kuykendall, Kiley, New York Institute of Technology College of Osteopathic Medicine at Arkansas State University, Jonesboro, Arkansas, United States
- Berman, Jonathan M., New York Institute of Technology College of Osteopathic Medicine at Arkansas State University, Jonesboro, Arkansas, United States
Background
Motivated by the desire to study the activity of proteases, and protease inhibitors and activators in urine, we are developing a novel surface plasmon resonance technique which enables monitoring of these reactions over a wide range of simultaneous substrate concentrations, facilitating high-throughput measurement of proteolysis against peptides from protease-regulated macromolecules such as Epithelial Sodium Channel (ENaC).
Methods
This study focuses on optimizing microfluidic chips for channel design, polydimethylsiloxane (PDMS) adhesion to glass, and gold-thin film sputter coating parameters while fabricating sensor chips. We developed a gradient generator chip design, which was cast from 3D-printed resin molds. BK7 glass slides were sputter coated with a gold thin-film. Both glass and PDMS were functionalized using oxygen plasma from a high voltage-high frequency generator to form covalent bonds.
Results
We systematically examined effects of gold coaching thickness, and deposition time on sensor chip functionality. Our results demonstrate successful gradient generation, and assembly of biomolecule monolayers on the gold surface.
Conclusion
Future work will integrate with the SPR design to measure real-world reaction kinetics, advancing our understanding of protease-substrate interaction and regulation in the kidney.