In this study, whey protein isolate-pullulan (WP/pullulan) microspheres were developed to entrap the probiotic Lactobacillus acidophilus NRRL-B 4495 by spray-drying technique. Microcapsules were analyzed for physicochemical characteristics including morphology, particle size, moisture content, water activity, dissolution time, and color properties. Results revealed that microcapsules were spherical in shape and obtained particle sizes between 5 and 160 lm, with an average size of around 50 lm. Blending pullulan with WP provided enhanced survival of probiotic bacteria during spray drying with a final viable cell number of 8.81 log CFU/g of microcapsule. Encapsulated probiotics were also found to have significantly (p 0.05) higher survived cell numbers compared to free probiotics under detrimental gastrointestinal conditions. Moreover, dissolution analysis suggested that protein-polysaccharide powdered microcapsules showed pH-sensitive dissolution properties in simulated gastric juice and simulated intestinal juice.