This study focused on generation of low-cost yet highly effective lithium selective bio-sorbent from olive pruning waste mainly olive branches. Olive branches were treated with NaOH that eliminated non-cellulosic materials and activated hydroxyl groups that inhibit the formation of active sites. Olive branches were then functionalized through homogeneous phosphorylation at 150 °C. POB, NOB, and FOB samples were subjected to SEM, XRD, FTIR, BET, XPS, and TGA to observe the changes in their structure and properties. Factors affecting lithium adsorption were investigated on the synthesized FOB in a batch system and analyzed by ICP-OES. Adsorption isotherms are well fitted to the Freundlich isotherm model than the Langmuir isotherm model which exhibited a maximum adsorption capacity of 6.7 mg/g at 30 °C. Kinetic studies exhibited fast kinetics and equilibrium was attained in 6 minutes while thermodynamic studies showed an exothermic, spontaneous reaction and increased randomness at the interaction interface. Regeneration studies proved the sustainability of FOB with Li+ desorption efficiency of 99.6% in 1.0 M HCl. The synthesized FOB displayed a better degree of column utilization and elution efficiency; 56.8% and 95.8% than Lewatit TP 260; 16.0% and 50.4% respectively in the adsorption column studies performed at room temperature. However, it exhibited a poor breakthrough capacity of 2.1 mg Li/ml sorbent than Lewatit TP 260 with 1.33 mg Li/ml sorbent. Based on all experimental results, the novel functionalized olive branches (FOB) proved a potential lithium selective bio-sorbent and can be applied in recovery of lithium from its aqueous sources.