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Biosensors
Volume 15
Issue 10
10.3390/bios15100643
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This is an early access version, the complete PDF, HTML, and XML versions will be available soon.
Article

Covalent and Site-Specific Immobilization of a Fluorogenic Sensor Protein on Cellulose-Based Paper for Detection of Lactate in Cell Culture Media

by
Ingo Bork
1,
Viktoria Höfling
1,
Janine Becker
1,
Markus Biesalski
2,
Tobias Meckel
2 and
Harald Kolmar
1,*
1
Institute for Organic Chemistry and Biochemistry, Technical University of Darmstadt, 64287 Darmstadt, Germany
2
Macromolecular and Paper Chemistry, Chemistry Department, Technical University of Darmstadt, 64278 Darmstadt, Germany
*
Author to whom correspondence should be addressed.
Biosensors 2025, 15(10), 643; https://doi.org/10.3390/bios15100643 (registering DOI)
Submission received: 11 August 2025 / Revised: 19 September 2025 / Accepted: 25 September 2025 / Published: 28 September 2025
(This article belongs to the Section Optical and Photonic Biosensors)
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Abstract

Lactate is a key metabolite with applications ranging from monitoring training efficiency to early sepsis detection and monitoring the metabolic state of cell cultures. In this study, a paper-based lactate sensor utilizing a fluorescent readout was developed. Unlike common lactate dehydrogenase (LDH)-based methods, these sensors use a green fluorescent protein (GFP) or mApple-coupled lactate binding domain, which provides a fluorescent readout upon lactate binding. We demonstrate that immobilizing these proteins on paper does not affect their ability to bind lactate and produce a fluorescent readout, by monitoring lactate levels in the cell culture supernatant applying different cell culture conditions.
Keywords: lactate sensor; fluorogenic sensor; paper-based sensor; covalent paper modification; site-specific immobilization lactate sensor; fluorogenic sensor; paper-based sensor; covalent paper modification; site-specific immobilization
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MDPI and ACS Style

Bork, I.; Höfling, V.; Becker, J.; Biesalski, M.; Meckel, T.; Kolmar, H. Covalent and Site-Specific Immobilization of a Fluorogenic Sensor Protein on Cellulose-Based Paper for Detection of Lactate in Cell Culture Media. Biosensors 2025, 15, 643. https://doi.org/10.3390/bios15100643

AMA Style

Bork I, Höfling V, Becker J, Biesalski M, Meckel T, Kolmar H. Covalent and Site-Specific Immobilization of a Fluorogenic Sensor Protein on Cellulose-Based Paper for Detection of Lactate in Cell Culture Media. Biosensors. 2025; 15(10):643. https://doi.org/10.3390/bios15100643

Chicago/Turabian Style

Bork, Ingo, Viktoria Höfling, Janine Becker, Markus Biesalski, Tobias Meckel, and Harald Kolmar. 2025. "Covalent and Site-Specific Immobilization of a Fluorogenic Sensor Protein on Cellulose-Based Paper for Detection of Lactate in Cell Culture Media" Biosensors 15, no. 10: 643. https://doi.org/10.3390/bios15100643

APA Style

Bork, I., Höfling, V., Becker, J., Biesalski, M., Meckel, T., & Kolmar, H. (2025). Covalent and Site-Specific Immobilization of a Fluorogenic Sensor Protein on Cellulose-Based Paper for Detection of Lactate in Cell Culture Media. Biosensors, 15(10), 643. https://doi.org/10.3390/bios15100643

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