Catalyst-free "click" functionalization of polymer brushes preserves antifouling properties enabling detection in blood plasma.

Progress in biosensors for clinical detection critically relies on modifications of the transducer surface to prevent non-specific adsorption from matrix components (i.e. antifouling) while supporting biomolecular recognition elements to capture the analyte. Such combination of properties presents a significant challenge. Hierarchically structured polymer brushes comprising an antifouling polymer bottom block and a functionalizable top block are proposed as a promising strategy to achieve this goal. We employed the catalyst-free strain-promoted alkyne-azide cycloaddition (SPAAC) "click" reaction to biofunctionalize antifouling polymer brushes without impairing their resistance to fouling. The functionalization was performed on the side chains along the top polymer block or only on the end-groups of the polymer brush. The immobilized amounts of bioreceptors (streptavidin followed by biotin-conjugated proteins) and the resistance to fouling from blood plasma of the surfaces obtained were evaluated via surface plasmon resonance. The end group functionalization approach resulted in very low immobilization of bioreceptor. On the other hand, the side group modification of a top polymer block led to immobilization of 83% of a monolayer of streptavidin. Following binding of a biotin-conjugated antibody (66 ng cm-2) the functionalized layer was able to reduce the fouling from undiluted human blood plasma by 89% in comparison with bare gold. Finally, the functionalized hierarchical polymer brushes were applied to the label-free detection of a model analyte in diluted human blood plasma, highlighting the potential for translation to medical applications.

Clickable Antifouling Polymer Brushes for Polymer Pen Lithography.

Protein-repellent reactive surfaces that promote localized specific binding are highly desirable for applications in the biomedical field. Nonspecific adhesion will compromise the function of bioactive surfaces, leading to ambiguous results of binding assays and negating the binding specificity of patterned cell-adhesive motives. Localized specific binding is often achieved by attaching a linker to the surface, and the other side of the linker is used to bind specifically to a desired functional agent, as e.g. proteins, antibodies, and fluorophores, depending on the function required by the application. We present a protein-repellent polymer brush enabling highly specific covalent surface immobilization of biorecognition elements by strain-promoted alkyne-azide cycloaddition click chemistry for selective protein adhesion. The protein-repellent polymer brush is functionalized by highly localized molecular binding sites in the low micrometer range using polymer pen lithography (PPL). Because of the massive parallelization of writing pens, the tunable PPL printed patterns can span over square centimeter areas. The selective binding of the protein streptavidin to these surface sites is demonstrated while the remaining polymer brush surface is resisting nonspecific adsorption without any prior blocking by bovine serum albumin (BSA). In contrast to the widely used BSA blocking, the reactive polymer brushes are able to significantly reduce nonspecific protein adsorption, which is the cause of biofouling. This was achieved for solutions of single proteins as well as complex biological fluids. The remarkable fouling resistance of the polymer brushes has the potential to improve the multiplexing capabilities of protein probes and therefore impact biomedical research and applications.