Controlled Peptide Capture and Release in 3D‐Printed Multimaterial Microstructures

High-resolution two-photon laser printing has revolutionized the fabrication of complex 3D micro- and nanostructures across a wide range of materials. However, the implementation of engineered biomolecules as functional, stimuli-responsive units remains underexplored. In this study, we present a method for fabricating 3D-printed hydrogel microstructures that contain de novo designed heterodimeric coiled-coil peptides to allow controlled peptide capture and release. Covalent incorporation of one coil strand into the 3D-printed network was combined with fluorescent labelling of the complementary strand. Controlling the assembly and disassembly of the coiled coil enabled selective binding and subsequent programmable release of the fluorescently labelled peptide using various external stimuli, such as pH, ionic strength, temperature, or peptide competitors. Using a set of orthogonal coiled-coil peptides, we fabricated multimaterial microstructures in which spatially resolved coiled-coil functionalization was achieved. Under complete spatiotemporal control, it was shown that the complementary coil strands could be captured and released selectively. This work demonstrates the use of discretely folded, chemically synthesizable peptides for 2PLP fabrication for the first time. Utilizing coiled-coil interactions as molecular handles enables the production of stimuli-responsive and reconfigurable hydrogel microstructures. This approach opens up possibilities for dynamic biomaterials, programmable drug delivery, and biochemical process engineering at the microscale.