Direct biophotolithographic method for generating substrates with multiple overlapping biomolecular patterns and gradients

We describe an approach to generate multicomponent surface-immobilized patterns and gradients on the basis of the photochemically controlled covalent coupling of solution-phase biomolecules to benzophenone-modified substrates. Gradients were simply achieved by continuously varying the exposure to nondamaging UV light across the surface with the gradient profile controlled by biomolecule concentration and the spatial and temporal illumination of the surface. Sequential exposure of the same surface in the presence of different biomolecules resulted in overlapping patterns and gradients of proteins and carbohydrates. Finally, we preliminarily demonstrate that the resulting surfaces are suitable for generating model substrates to probe cell-substrate interactions.