The feasibility of alleviating burst release of electrospun bead-on-string nanofiber scaffolds loaded with protein particles was evaluated, including an investigation of the influence of the beads number on the release profile. Bovine serum albumin-loaded dextran particles were used as the model drug and poly(lactic-co-glycolic acid) as the polymer to fabricate the bead-on-string nanofiber scaffolds by electrospinning. Both the bead structure and the distribution of the particles in the beads were examined by scanning electron, transmission electron, and fluorescence microscopy. The results of fluorescence microscopy suggested that the particles were well encapsulated by the beads of the fibers. In vitro release tests showed that a more sustainable release profile with less initial burst release could be obtained from the bead-on-string fibers than from smooth fibers with uniform diameter. In addition, when the number of the forming beads was not numerous enough to encapsulate all the particles in the suspensions, the release performance worsened because the surplus particles were not properly encapsulated.