| Autor: |
Riedl V; Institute of Biochemistry, Leipzig University, Johannisallee 21-23, 04103, Leipzig, Germany., Heiser L; Institute of Biochemistry, Leipzig University, Johannisallee 21-23, 04103, Leipzig, Germany., Portius M; Institute of Biochemistry, Leipzig University, Johannisallee 21-23, 04103, Leipzig, Germany., Schmidt JO; Institute of Biochemistry, Leipzig University, Johannisallee 21-23, 04103, Leipzig, Germany., Pompe T; Institute of Biochemistry, Leipzig University, Johannisallee 21-23, 04103, Leipzig, Germany. |
| Abstrakt: |
Sulfonamide antibiotics were the first synthetic antibiotics on the market and still have a broad field of application. Their extensive usage, wrong disposal, and limited degradation technologies in wastewater treatment plants lead to high concentrations in the environment, resulting in a negative impact on ecosystems and an acceleration of antibiotic resistance. Although lab-based analytical methods allow for sulfonamide detection, comprehensive monitoring is hampered by the nonavailability of on-site, inexpensive sensing technologies. In this work, we exploit functionalized elastic hydrogel microparticles and their ability to easily deform upon specific binding with enzyme-coated surfaces to establish the groundwork of a biosensing assay for the fast and straightforward detection of sulfonamide antibiotics. The detection assay is based on sulfamethoxazole-functionalized hydrogel microparticles as sensor probes and the biomimetic interaction of sulfonamide analytes with their natural target enzyme, dihydropteroate synthase (DHPS). DHPS from S. pneumoniae was recombinantly produced by E. coli and covalently coupled on a glass biochip using a reactive maleic anhydride copolymer coating. Monodisperse poly(ethylene glycol) hydrogel microparticles of 50 μm in diameter were synthesized within a microfluidic setup, followed by the oriented coupling of a sulfamethoxazole derivative to the microparticle surface. In proof-of-concept experiments, sulfamethoxazole, as the most used sulfonamide antibiotic in medical applications, was demonstrated to be specifically detectable above a concentration of 10 μM. With its straightforward detection principle, this assay has the potential to be used for point-of-use monitoring of sulfonamide antibiotic contaminants in the environment. |
