The transport and fate of antibiotics are significantly influenced by co-existing colloidal and nanosized substances, such as clay particles. Montmorillonite, a common clay mineral with a thin nano-sheet-like structure, enhances antibiotic (e.g., ciprofloxacin) mobility due to its strong adsorption properties. Nevertheless, little is known about how ciprofloxacin systematically influences the charging and aggregation properties of montmorillonite. This study examines the effect of ciprofloxacin on the electrophoretic mobility and hydrodynamic diameter of Na-montmorillonite under varying pH levels and NaCl concentrations. Results show ciprofloxacin promotes aggregation and alters the surface net charge of Na-montmorillonite at acidic to neutral pH, where ciprofloxacin is positively charged. At higher pH levels, where ciprofloxacin is negatively charged, no significant effects are observed. The observed aggregation behaviors align with predictions based on the Derjaguin-Landau-Verwey-Overbeek (DLVO) theory. Specifically, the slow aggregation regime, the fast aggregation regime, and the critical coagulation concentration are identified. The relationship between critical coagulation ionic strength and electrokinetic surface charge density is well explained by the DLVO theory with the Debye-Hückel approximations. Additionally, non-DLVO interactions are inferred. At low NaCl and ciprofloxacin concentrations, minimal changes in aggregation and surface charge suggest dispersed montmorillonite may facilitate ciprofloxacin transport, raising environmental concerns.