Disclosed is a peptide-immobilized substrate for measuring a target protein, with which the peptide can have a structure required for being recognized by the target protein, with which the accurate loading amount of the peptide can be attained, and by which a trace amount of the target protein may be measured accurately and simply. The peptide-immobilized substrate for measuring a target protein according to the present invention comprises a chemically synthesized peptide having an expected spatial structure or having a binding ability with the target protein, which peptide can bind with the target protein and is immobilized on the substrate.