The application of cryogels for biomolecule purification has expanded due to their adsorption efficiency and operational advantages. In this study, polyacrylamide cryogels functionalized with L-phenylalanine (cryogel-Phe) via the glutaraldehyde method were designed for lysozyme adsorption. Cryogel functionalization was confirmed by Fourier-transform infrared spectroscopy and Kjeldahl analysis, indicating the immobilization of 458.65 mg phenylalanine g cryogel À1 . Cryogel-Phe showed high porosity (0.95) and a Young's modulus of 526.71 kPa. Thermogravimetric analysis indicated that thermal degradation occurred above 200 C. Differential scanning calorimetry and X-ray diffraction confirmed that the cryogel material was amorphous. In addition, the column presented a hydraulic permeability of 4.15 Â 10 À13 m 2 , axial dispersion ranging from 10 À7 to 10 À6 m 2 s À1 , and a height equivalent to a theoretical plate ranging from 0.10 to 0.21 cm. The highest adsorption of lysozyme (67.65 mg g À1 ) was obtained using sodium thiocyanate saline solution (0.025 mol L À1 , pH 5.0). The ability of the cryogel-Phe column to capture and purify lysozyme was confirmed by high enzymatic activity (1294.17 U ml À1 ), purity (87.92%), purification factor (11.49), and sulphate-polyacrylamide electrophoresis gel (SDS-PAGE) electrophoresis gel.