The bioreceptor immobilization process (biofunctionalization) turns to be one of the bottlenecks when developing a competent and high sensitivity label-free biosensor. Classical approaches seem to be effective but not efficient. Although biosensing capacities are shown in many cases, the performance of the biosensor is truncated by the inefficacious biofunctionalization protocol and the lack of reproducibility. In this work, we describe a unique biofunctionalization protocol based on chemical surface modification through silane chemistry on SiO2 optical sensing transducers. Even though silane chemistry is commonly used for sensing applications, here we present a different mode of operation, applying an unusual silane compound used for this purpose (3-Ethoxydimethylsilyl)propylamine, APDMS, able to create ordered monolayers, and minimizing fouling events. To endorse this protocol as a feasible method for biofunctionalization, we performed multiple surface characterization techniques after all the process steps: Contact angle (CA), X-ray photoelectron spectroscopy (XPS), ellipsometry, and fluorescence microscopy. Finally, to evidence the outputs from the SiO2 surface characterization, we used those SiO2 surfaces as optical transducers for the label-free biosensing of matrix metalloproteinase 9 (MMP9). We found and demonstrated that the originally designed protocol is reproducible, stable, and suitable for SiO2-based optical sensing transducers.
Keywords: biofunctionalization; label free biosensor; surface characterization; surface modification.