As starch derivatives, beta-cyclodextrin (beta-CD) has received increasing attention due to its special chemical structure and excellent properties. In this study, a novel beta-CD based on N-halamine antimicrobial copolymer (beta-CD-MAH-VBDMH) was synthesized and characterized by solid-state C-13 NMR and GPC. The electrospun nanofibers were obtained using mixed solution of beta-CD-MAH-VBDMH and cellulose acetate (CA). The obtained nanofibers were characterized by SEM, XPS, XRD, TGA, and DSC. After chlorination treatment by sodium hypochlorite, biocidal efficacies of chlorinated nanofibers against Escherichia coli 0157:H7 (ATCC 43895) and Staphylococcus aureus (ATCC 6538) were evaluated using a modified AATCC 100-1999 method. The fibers showed excellent antimicrobial properties against the used two bacteria within brief contact times. OVA irradiation tests indicated that the chlorine bound to the fibers is relatively stable, and most of the lost chlorine could be recovered after re-chlorination process. The mouse 3T3 fibroblasts in vitro cell cytocompatibility studies demonstrated that the antibacterial nanofibers have good biocompatibility. (C) 2014 Elsevier B.V. All rights reserved.