Virus-Assembled Biofunctional Microarrays with Hierarchical 3D Nano-Reticular Network

Three-dimensional (3D) hierarchical wrinkled materials built with biological entities have so far remained exclusive to nature. Herein, multiscale functional ultraporous 3D bio-networks of bioprinted phage-built wrinkled microarrays are created through establishing a universal heat- and solvent-independent substrate-shrinkage method induced by high-pressure carbon dioxide (HPCD). This method results in diverse wrinkled patterns on soft materials and is particularly powerful for solvent- and heat-sensitive biomaterials, for which other methods have failed. The phage nanofilaments (7 nm width) self-assemble into orderly-aligned submicron bundles (100 nm width), which crimp into tunable microscale wrinkles (0.7–5.0 µm width) on size-controllable micro-arrays (200–600 µm width) exhibiting a four-level hierarchical nano-reticular structure. The HPCD method also protects the bioactivity of biorecognition molecules loaded into the microarrays, leading to the design of bacteria-sensing chips, made with in-house deoxyribozyme-loaded 3D phage microarrays. The developed bacteria-sensing chips achieve a limit of detection that is 100 × more sensitive with greater reproducibility compared to two-dimensional (2D) microdot arrays and correctly identify Legionella pneumophila in contaminated water samples collected from industrial cooling towers, highlighting phage-built wrinkled networks as a platform for bottom-up assembly of biological building blocks into biofunctional material.