Lithographing of biomolecules on a substrate surface using an enzyme-immobilized AFM tip

A method of enzymatic lithography was successfully performed in a buffered solution using Staphylococcal serine V8 protease and atomic force microscopy (AFM). The retained activity of the protease immobilized on the AFM tip was confirmed by force measurement to rupture the enzyme-substrate complex. After contact scanning using the enzyme-immobilized tip to the substrate peptide layer, a square shape of the lithographed area was clearly observed by subsequent AFM imaging.