The deposition of biomolecules on biosensing surface platforms plays a key role in achieving the required sensitivity and selectivity for biomolecular interactions analysis. Controlling the interaction between the surface and biomolecules is increasingly becoming a crucial design tool to modulate the surface properties needed to improve the performance of the assay and the detection outcome. Carboxymethyl-dextran (CMD) coating can be exploited to promote chemical grafting of proteins, providing a hydrophilic, bioinert, nonfouling surface and a high surface density of immobilized proteins. In the present work, we developed and optimized a technique to produce a cost-effective CMD-based patterned surface for the immobilization of biomolecules to be used on standard protocols optimization. They consist of silicon or glass substrates with patterned bioactive areas able to efficiently confine the sampling solution by simply exploiting hydrophilic/hydrophobic patterning of the surface. The fabrication process involves the use of low-cost instruments and techniques, compatible with large scale production. The devices were validated through a chemiluminescence assay we recently developed for the analysis of binding of DNA nanoassemblies modified with an affinity binder to target proteins immobilized on the bioactive areas. Through this assay we were able to characterize the chemical reactivity of two target proteins toward a dextran matrix on patterned surfaces and to compare it with model CMD-based surface plasmon resonance (SPR) surfaces. We found a high reproducibility and selectivity in molecular recognition, consistent with results obtained on SPR sensor surfaces. The suggested approach is straightforward, cheap, and provides the means to assess patterned functionalized surfaces for bioanalytical platforms.
Keywords: binding bioassay; carboxymethyl-dextran; patterned surfaces; protein anchoring; surface plasmon resonance.