Easily Accessible Protein Nanostructures via Enzyme Mediated Addressing

Abstract

Site-specific formation of nanoscaled protein structures is a challenging task. Most known structuring methods are either complex and hardly upscalable or do not apply to biological matter at all. The presented combination of enzyme mediated autodeposition and nanosphere lithography provides an easy-to-apply approach for the buildup of protein nanostructures over a large scale. The key factor is the tethering of enzyme to the support in designated areas. Those areas are provided via prepatterning of enzymatically active antidots with variable diameters. Enzymatically triggered protein addressing occurs exclusively at the intended areas and continues until the entire active area is coated. After this, the reaction self-terminates. The major advantage of the presented method lies in its easy applicability and upscalability. Large-area structuring of entire support surfaces with features on the nanometer scale is performed efficiently and without the necessity of harsh conditions. These are valuable premises for large-scale applications with potentials in biosensor technology, nanoelectronics, and life sciences.

Enzyme‐Mediated In Situ Buildup and Site‐Specific Addressing of Polymeric Coatings

“…Only very few stranded particles are found beyond the area of enzyme functionalization. This result moreover suggests adaptability of the nanostructuring ability of EMA, which was previously shown for biological particles, to synthetic polymers. However, experimental proof of this was not within the scope of this work but will be addressed in future experiments.…”

“…Additionally, this tool enables site-specific deposition, including the formation of patterns, down to real nanostructures. [16] Figure 1 shows a sketch of the process.…”

“…This inherent specificity of the enzyme‐mediated addressing ensures that the polymerization reaction occurs exclusively within the reaction zone. Additionally, this tool enables site‐specific deposition, including the formation of patterns, down to real nanostructures . Figure shows a sketch of the process.…”

Site-Specific Addressing of Particles and Coatings via Enzyme-Mediated Destabilization

“…Performed in bulk, this reaction is long known for the sake of milk clotting. Performed within the specifics of the EMA, it is highly suitable for the production of defined protein structures on arbitrary surfaces, e.g., for applications in life sciences or adhesion [12,15].…”

“…Application of high ionic strength in the immobilization buffer is known to be a feasible tool to drive favorable orientation to a certain degree [32]. In the context of EMA this level of control has proven to be sufficient, even for the production of genuine nano-structures over a macroscopic scale [15]. Therefore, this technique is also used in this work.…”