We investigated the effect of the major process parameters (crystallization temperature and time) on the efficiency of the vancomycin crystallization process and conducted a kinetic and thermodynamic analysis. The most clear and uniform vancomycin crystals with the highest yield (~98%) were obtained at the optimum crystallization temperature (283 K) and time (1,440 min). The electron microscope, SEM, and XRD analyses showed that intact crystalline vancomycin was obtained when using a crystallization temperature of 283, 288, and 293 K. The kinetic analysis results revealed that the Johnson-Mehl-Avrami-Kolmogorov (JMAK) model was suitable with a high value for r2 (>0.9561) and low value for RMSD (<0.0170). Finally, from the thermodynamic analysis the Gibb’s free energy change (ΔG0), entropy change (ΔS0), and enthalpy change (ΔH0) were all negative, indicating that the crystallization process was spontaneous, irreversible, and exothermic.