In the biosensing platform, label-free detection technique provides advantages such as the short analysis time and the cost-effectiveness. In this study, we showed the feasibility of the LSPR substrate with gold nanoparticle array for detecting low density lipoprotein (LDL) and high density lipoprotein (HDL) without labeling. The LSPR substrate was fabricated through the lift-off process with the anodized alumina mask, and its LSPR phenomenon was observed by measuring the optical transmission of substrate. The antibodies were immobilized on the gold nanoparticle array via the chemical binding, in which the 11-MUA was used as the linker to bind the antibodies. The binding of antibodies was confirmed by observing the shift of LSPR peak of the substrate. Finally, with the LSPR substrates with the antibodies immobilized, the detection of LDL and HDL was investigated. As a result, LDL and HDL could be detected in the clinically available concentration range, respectively.