Chitosan-supported; Ciprofloxacin-HCl-Chitosan/Tween/Tri polyphosphate (CIP@CH-TW/TPP) an efficient drug delivery system has been successfully developed using simple sol-gel chemical route. The drug encapsulated nanocomposite was interacted with bovine serum albumin (BSA) to deliver the drug at targeted site. Incorporation of tween-80 onto CIP/CH-TPP enhanced loading capacity of nanocomposite and improves drug releasing efficiency up to 61–78% as compared to native chitosan (37%). The CH-TW/TPP nanocomposite has been characterized using FTIR, TGA-DTA, DLS, SEM and XRD analyses. Biophysical properties were performed to achieve optimum interaction of CIP@CH-TW/TPP with BSA that is the novelty of the present study. The effect of surfactant concentration on CIP–BSA interaction was studied using different concentrations of Tween-80 (1 μM and 2 μM) via fluorescence, UV–Vis and circular dichroism (CD) technique. The findings of fluorescence quenching study reveal the binding of CIP with BSA via a static process which results formation of CIP–BSA complex. The results of thermodynamic studies (ΔG, ΔH and ΔS) indicate that the hydrogen bonding and Vander Waals forces play a vital role to achieve optimal CIP-BSA interaction. Molecular modeling calculations and displacement study results confirm the binding of CIP with BSA at site I. Significant results of drug–protein interactions establish the applicability of synthesized CIP@CH-TW/TPP nanocomposite in drug delivery system. Display omitted •Development of Ciprofloxacin-HCl loaded CH/TW-TPP nanocomposite for drug delivery•The interactions of CIP and NCIP with BSA using biophysical methods and molecular docking•Improvement in clinical efficacy of ciprofloxacin using CH/TW-TPP nanocomposite