Keywords

biosensor, cell-free protein synthesis (CFPS), glutamine, aspartate, cancer

Abstract

Glutamine is an essential biomolecule that plays a pivotal role in many diseases, such as cancer, where it can serve as fuel for rapid proliferation. Treatments for these diseases can be monitored and optimized through the detection of glutamine, though standard glutamine detection procedures are costly and require complex instrumentation. Cell-free protein synthesis (CFPS) has recently enabled a paper-based, colorimetric glutamine sensor that carries the potential to increase test accessibility while dramatically reducing consumer cost to enable at-home, rapid treatment monitoring. Test sensitivity remained limited by residual assay background, thus motivating this work where CFPS reactions traditionally formulated with glutamate salts were compared to systems using alternative salts, including aspartate, acetate, citrate, and sulfate, to reduce the background generation of glutamine. This led to the discovery of a novel aspartate-based CFPS system that boasts a high signal strength and indetectable background noise over 225 min. Acetate-, citrate-, and sulfate-based systems also yielded zero background glutamine detection but at a lower signal response compared to the aspartate-based system. These findings mark crucial advancements in producing a cost-effective, simple glutamine monitor while simultaneously showcasing the adaptability of CFPS’s open reaction environment for solving complex challenges in next-generation biosensor development.

Original Publication Citation

Talley, J. P., Free, T. J., Green, T. P., Chipman, D. M., & Bundy, B. C. (2025). Eliminating Assay Background of a Low-Cost, Colorimetric Glutamine Biosensor by Engineering an Alternative Formulation of Cell-Free Protein Synthesis. Chemosensors, 13(6), 206. https://doi.org/10.3390/chemosensors13060206

Document Type

Peer-Reviewed Article

Publication Date

2025-06-05

Publisher

Chemosensors

Language

English

College

Ira A. Fulton College of Engineering

Department

Chemical Engineering

University Standing at Time of Publication

Full Professor

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