This paper describes the preparation of the metallic affinity column by atom transfer radical polymerization of glycidyl methacrylate (GMA) on Wang resin surface. The metallic ions (Zr 4þ , Fe 3þ , and Pd 2þ ) were introduced after phosphonation of the epoxy group of the grafted poly(GMA) on Wang resin surface. The successful results on metallic ion-immobilized polymeric microsphere stationary phase were confirmed via Fourier transform infrared, scanning electron microscopy, X-ray photoelectron spectra (XPS), and inductively coupled plasma-atomic emission spectrometer. The resolution degree of the phosphonated casein and dephosphonated casein for Zr 4þ , Fe 3þ , and Pd 2þ affinity column was examined. The result shows that the resolution time (min) of the phosphonated casein was higher than that of the dephosphated casein on the Zr 4þ affinity column by liquid chromatography. However, the phosphonated casein and dephosphonated casein were not separated on the Fe 3þ affinity column and Pd 2þ affinity column by liquid chromatography.