A method for forming a molecularly imprinted polymer biosensor includes: (a) preparing a reaction solution including an imprinting molecule, a functional monomer, an initiator, and a crosslinking agent (b) disposing the reaction solution in a space between upper and lower substrates each of which is made of a light-transmissible material (c) disposing on the upper substrate a photomask having a patterned hole (d) irradiating the reaction solution through the patterned hole of the photomask and the upper substrate so that the reaction solution undergoes polymerization to form a polymer between the upper and lower substrates (e) removing the upper substrate after the polymer is formed on the lower substrate and (f) extracting the imprinting molecule from the polymer so that a patterned molecularly imprinted polymer film is formed on the lower substrate.
