Nanoparticles (NPs) have been developed as an important strategy to deliver low molecular-weight drugs, as well as biomacromolecules such as proteins or DNA. The body distribution of colloidal drug delivery systems was mainly influenced by two physicochemical properties namely particle size and surface characteristics. Particle size is a crucial parameter, in particular for the in vivo behavior of NPs after intravenous injection. The objective of the present study was the preparation of human serum albumin (HSA) nanoparticle (NP) by desolvation method and optimization of NP by applying the Taguchi method together with characterization of the NP bioproducts for drug delivery application. Several process parameters were examined to achieve a suitable size of NP such as pH, HSA concentration, organic solvent adding rate and the ratio of organic solvent/HSA solution. Taguchi method with L 16 orthogonal array robust design was implemented to optimize experimental conditions of the purpose. This approach facilitates the study of interaction of a large number of variables spanned by factors and their settings with a small number of experiments leading to considerable saving in time and cost for the process optimization. As a result of Taguchi analysis in this study, pH and ratio of organic solvent/HSA solution were the most influencing -1 parameters on the particle size. The minimum size of NPs (53 nm) were obtained at pH 9, 75 mg.ml HSAconcentration, ratio of organic -1 solvent/HSA solution of 4 and organic solvent adding rate of 1.5 ml.min . The mechanistic of the optimum conditions for preparing protein NPs and their characterization as drug delivery vehicles are discussed

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