The preparation of a fluorine-containing synergistic nonfouling/fouling-release surface, using a b-PFMA-PEO asymmetric molecular brush possessing both poly(ethylene glycol) (PEO) and poly(2,2,2-trifluoroethyl methacrylate) (PFMA) side chains densely distributed on the same repeat unit along the polymeric backbone, is reported. On the basis of the poly(Br-acrylate-alkyne) macroagent comprising two functionalities (alkynyl and 2-bromopropionate), which is prepared by reversible addition-fragmentation chain transfer homopolymerization of a new trifunctional acrylate monomer of Br-acrylate-alkyne, b-PFMA-PEO asymmetric molecular brushes are obtained by concurrent atom transfer radical polymerization and Cu-catalyzed azide/alkyne cycloaddition "click" reaction in a one-shot system. A spin-cast thin film of the b-PFMA-PEO asymmetric molecular brush exhibits a synergistic antifouling property, in which PEO side chains endow the surface with a nonfouling characteristic, whereas PFMA side chains display the fouling-release functionality because of their low surface energy. Both protein adsorption and cell adhesion tests provided estimates of the antifouling activity of the asymmetric molecular brush surfaces, which was demonstrated to be influenced by the degree of polymerization of the backbone and the length of the PEO and PFMA side chains. With compositional heterogeneities, all asymmetric molecular brush surfaces show considerable antifouling performance with much less protein adsorption (at least 45% off, up to 75% off) and cell adhesion (at least 70% off, up to 90% off) in comparison with a bare surface.