In studying biological interfaces where carbohydrates are prevalent, scientists are only beginning to investigate how chemical composition, binding thermodynamics, and 3D structure work synergistically to control development, communication, and disease progression. Capturing the interfacial dynamics that drive the mechanical and biological properties of natural carbohydrate-based nanomaterials will require synergistic advances in 3D nanolithography, surface characterization, and organic and macromolecular chemistry at interfaces. This Faraday Discussion brings together researchers from various fields including printing tool development, surface chemistry, binding thermodynamics, and glycobiology to discuss how 3D nanolithography can increase our understanding of biointerfaces.