The currently available purification protocol for human papillomavirus (HPV) pseudovirus (PsV), as a substitute for the native HPV virion, utilizes Optiprep gradients (OG), which require costly equipment such as ultracentrifuges, and 4-7 h of working time, and cannot cope with large PsV samples. To develop a convenient method for purifying HPV type 58 PsV (HPV58 PsV) we have examined the use of heparin chromatography (HC) and cation-exchange chromatography (CC), which utilize open column systems and do not require expensive equipment. We confirmed that the PsVs resulting from HC and CC have correctly assembled conformations and are neutralized by anti-HPV58 PsV mouse serum, indicating that their antigenic characteristics make them suitable to substitute for native HPV58 virion. The recoveries of infectious PsV resulting from HC and CC were 39% and 11%, respectively, while that from OG was 58%. The two new purification methods are advantageous with respect to working time (only 30 min) and expense over the OG method, and have considerable potential for large scale purification. (C) 2010 Elsevier Inc. All rights reserved.