In this study, we describe the development of a cancer biomarker-sensitive nanobiosensor based on localized surface plasmon resonance that enables recognition for proteolytic activity of membrane type 1 matrix metalloproteinase (MT1-MMP) anchored on invasive cancer cells. First of all, we prepared biomarker-detectable substrate based on gold nanorods (GNRs) using nanoparticle adsorption method. The sensitivity of the sensing chip was confirmed using various solvents that have different refractive indexes. Subsequently, MT1-MMP–specific cleavable peptide was conjugated onto the surface of GNRs, and molecular sensing about proteolytic activity was conducted using MT1-MMP and cell lysates. Collectively, we developed a biomarker detectable sensor, which allows for the effective detection of proteolytic activity about MT1-MMP extracted from invasive cancer cells.