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Advances in carbohydrate sequencing technologies have revealed the tremendous complexity of the glycome. This complexity reflects the structural and chemical diversity of carbohydrates and is greater than that of proteins and oligonucleotides. The next step in understanding the biological function of carbohydrates requires the identification and quantification of carbohydrate interactions with other biomolecules, in particular, with proteins. To this end, we have developed a cantilever array biosensor with a self-assembling carbohydrate-based sensing layer that selectively and sensitively detects carbohydrate-protein binding interactions. Specifically, we examined binding of mannosides and the protein cyanovirin-N, which binds and blocks the human immunodeficiency virus (HIV). Cyanovirin-N binding to immobilized oligomannosides on the cantilever resulted in mechanical surface stress that is transduced into a mechanical force and cantilever bending. The degree and duration of cantilever deflection correlates with the interaction's strength, and comparative binding experiments reveal molecular binding preferences. This study establishes that carbohydrate-based cantilever biosensors are a robust, label-free, and scalable means to analyze carbohydrate-protein interactions and to detect picomolar concentrations of carbohydrate-binding proteins.

For the miniaturization of biological assays, especially for the fabrication of microarrays, immobilization of biomolecules at the surfaces of the chips is the decisive factor. Accordingly, a variety of binding techniques have been developed over the years to immobilize DNA or proteins onto such substrates. Most of them require rather complex fabrication processes and sophisticated surface chemistry. Here, a comparatively simple immobilization technique is presented, which is based on the local generation of small spots of surface attached polymer networks. Immobilization is achieved in a one-step procedure: probe molecules are mixed with a photoactive copolymer in aqueous buffer, spotted onto a solid support, and cross-linked as well as bound to the substrate during brief flood exposure to UV light. The described procedure permits spatially confined surface functionalization and allows reliable binding of biological species to conventional substrates such as glass microscope slides as well as various types of plastic substrates with comparable performance. The latter also permits immobilization on structured, thermoformed substrates resulting in an all-plastic biochip platform, which is simple and cheap and seems to be promising for a variety of microdiagnostic applications.