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

Abstract 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.