In this work, the performance and adsorption characteristics of macroporous resins for the recovery and enrichment of ganoderic acid (GA)-Mk and GA-T from Ganoderma lucidum mycelia were systematically evaluated. ADS-8 resin displayed the best adsorption and desorption capacities among the tested resins based on batch experiments. The interaction between solute and ADS-8 resin at different temperatures was described in terms of Langmuir and Freundlich isotherms, and the equilibrium experimental data were well fitted to the two isotherms. Thermodynamic analysis indicated the exothermic and spontaneous nature of the adsorption process. The adsorption capacity of ADS-8 resin was found to depend strongly on the pH value of the initial solution. Dynamic adsorption and desorption tests were performed on an ADS-8 resin-packed column to obtain optimal parameters for recovering GA-Mk and GAT from G. lucidum extract. Under optimized conditions, a laboratory scale-up preparation of GA-Mk and GA-T was carried out. The contents of GA-Mk and GA-T were increased from 45 to 22 mg/g in the crude extract to 352 and 141 mg/g in the final product with recovery yields of 90.1 and 72.2%, respectively. These results demonstrated that ADS-8 resin chromatography could act as a useful approach for obtaining ganoderic acids from G. lucidum mycelia.