Abstract: Bovine serum albumin (BSA) molecularly imprinted polymer (MIP) surface plasmon resonance (SPR) biosensors were prepared using BSA as the model, dopamine as the functional monomer and cross-linker and sodium periodate as the oxidant. BSA molecules were fixed in advance through hydrogen bond via introducing poly(acrylic acid) (PAA) to increase recognition sites, and partially hydrolyzed poly(2-methyl-2-oxazoline)(PMOXA-EI) was also introduced into the noncavity regions of polydopamine to resist nonspecific adsorption of proteins. The prepared BSA-MIP SPR sensor was characterized by X-ray photoelectron spectroscopy, atomic force microscopy, variable angle spectroscopic ellipsometry, and static water contact angle. SPR adsorption studies were carried out in aqueous BSA solutions with the mass concentration of 0.1—10 μg/mL. The limit of detection and the limit of quantification values were obtained as 53 ng/mL and 161 ng/mL, respectively. Selectivity studies were performed against β-lactoglobulin, ovalbumin, lysozyme, and cytochrome C, and the corresponding selectivity coefficients were determined to be 4.43, 3.45, 3.17 and 3.64, respectively. Finally, BSA-MIP biosensor was used to detect BSA in the mixture solution of the above five proteins. The BSA recovery rate was 97.5%—102.5%.