A macroporous monolithic material based on an N-hydroxyphthalimide ester of acrylic acid-co-glycidyl methacrylate-co-ethylene dimethacrylate terpolymer was synthesized by photoinitiated free-radical polymerization. Several porogenic solvents, such as cyclohexanol, dodecanol, and poly(ethylene glycol)s, were tested to obtain the monolithic material with an optimal pore size allowing unrestricted penetration of large molecule (proteins) into a three-dimensional porous space. The new monolithic material was covalently bound to an inert surface (glass) directly in the polymerization step, and it was suggested as a solid matrix for the development of new types of three-dimensional protein microarrays (biochips). A demonstration of the potential of the suggested microarray platform as well as optimization of microarray performance conditions was realized with a model mouse immunoglobulin G/goat anti-mouse imunoglobulin G affinity pair. (C) 2008 Wiley Periodicals, Inc. J Appl Polyin Sci 111:692-700,2009