A quaternized cotton linter fiber (QCLF) based adsorbent for removal of phosphate was prepared by grafting glycidyl methacrylate onto cotton linter and subsequent ring-opening reaction of epoxy groups and further quaternization. The adsorption behavior of the QCLF for phosphate was evaluated in a batch and column experiment. The batch experiment demonstrated that the adsorption process followed pseudo-second-order kinetics with an R2 value of 0.9967, and the Langmuir model with R2 value of 0.9952. The theoretical maximum adsorption capacity reached 152.44 mg/g. The experimental data of the fixed-bed column were well fitted with the Thomas and Yoon-Nelson models, and the adsorption capacity of phosphate at 100 mg/L and flow rate 1 mL/min reached 141.58 mg/g. The saturated QCLF could be regenerated by eluting with 1 M HCl.