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Measurement of Protein Kinetics Using a Liquid Phase-Based Biosensing Platform
Edwin J Ortiz-Riaño1, Diana L Mancera-Zapata1, Martha Ulloa-Ramírez1,2
1Biophotonic Nanosensors Laboratory, Centro de Investigaciones en Óptica, A. C., Loma del Bosque 115, Lomas del Campestre, León, Guanajuato37150, Mexico.
Analytical Chemistry
|October 17, 2022
Summary
This study introduces a simple graphene oxide biosensor for real-time protein kinetics analysis. The platform offers an affordable and effective alternative to traditional methods for determining protein-binding constants.
Area of Science:
- Biomedical Sciences
- Biochemistry
- Analytical Chemistry
Background:
- Macromolecular associations are vital in biomedical fields like drug development and diagnostics.
- Traditional methods for determining protein kinetics, such as surface plasmon resonance, are costly and complex.
Purpose of the Study:
- To develop a simple, real-time biosensing platform for readily determining protein-binding constants.
- To offer an affordable and effective alternative to existing protein kinetics analysis methods.
Main Methods:
- Utilized graphene oxide-modified microwell plates and fluorophore-labeled proteins for real-time detection.
- Employed fluorescence quenching by graphene oxide to differentiate bound and unbound probes.
- Validated the platform using three protein pairs: H-IgG, PSA, and SARS-CoV-2 spike protein RBD.
Main Results:
- Demonstrated successful determination of protein-binding constants using the graphene oxide platform.
- Achieved comparable and consistent results with existing methods and literature data.
- Developed open-source software for automated determination of binding kinetics constants.
Conclusions:
- The developed platform provides a simple, effective, and economical method for protein kinetics analysis.
- This approach can be performed using standard microwell plate readers and inexpensive materials.
- Envisioned as a foundation for a new generation of affordable diagnostics.
