Polymer nanoparticles have been widely used in many biomedical applications, constituting a major incentive for immobilization of proteins. Poly(methyl methacrylate) nanoparticles were synthesized through miniemulsion polymerizations and used as supports for bovine serum albumin immobilization. Particularly, the effects of surfactant type (anionic sodium dodecyl sulfate and cationic cetyl trimethyl ammonium bromide) surfactant concentration and monomer holdup on some of the final nanoparticle properties (particle sizes, zeta potential and protein load) were characterized with help of statistical experimental designs for the first time. Results showed that the characteristics of the surfactant controlled the BSA adsorption efficiency, with enhanced rates of adsorption on the anionic particle surfaces, showing that the surfactant exerts fundamental effect on functionalization of emulsified polymer particles, which must be explicitly acknowledged in studies of polymer particle functionalization with proteins. Finally, BSA adsorption was shown to follow a multilayer process, given the better fitting with the Freundlich model.