Hepatitis B surface antibody (HBsAb) imprinted poly(hydroxyethyl methacrylate-N-methacryloyl-L-tyrosine methyl ester) (PHEMAT) film on the surface plasmon resonance (SPR) sensor chip was prepared for diagnosis of HBsAb in human serum. Gold SPR chip surface was modified with allyl mercaptane and, then, HBsAb-imprinted PHEMAT film was formed on the chip surface. Surface characterization of the non-modified, allyl mercaptane modified and HBsAb-imprinted PHEMAT SPR chips were investigated with contact angle, atomic force microscopy (AFM). Kinetic studies were performed using HBsAb positive human serum. In order to determine the kinetic and binding constants, Scatchard, Langmuir, Freundlich and Langmuir-Freundlich models were applied to experimental data. Scatchard curve shows that HBsAb imprinted SPR chip has some surface heterogeneity, SPR chip obeyed the Langmuir adsorption model. The maximum detection limit was 208.2 mIU/mL. K(A) and K(D) values are 0.015 mIU/mL and 66.0 mL/mIU, respectively. Control experiments of the SPR chip were performed using non-immunized, HBsAb negative serum. The control experiment results show that SPR chip does not give any noticeable response to HBsAb negative serum.