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.