Peptides enable the construction of a diversity of one-dimensional (1D) and zero-dimensional (0D) nanostructures by molecular self-assembly. To date, it is a great challenge to construct two-dimensional (2D) nanostructures from peptides. Here we introduce an organic molecule to tune the amphiphilic-like peptide assembly to form a peptide-organic 2D nanopatch structure. The nanomechanical properties of the nanopatch were explored by quantitative nanomechanical imaging and force control manipulation. The peptide-organic patches are multilayers composed of several domains, which can be peeled off stepwise. The patch formation provides an approach towards constructing 2D nanostructures by peptide-organic assembly and it could be potentially utilized in a wide range of applications such as functional biomaterials.