Comparison of Methods for The Purification of Goat Lactoferrin and Antiviral Activity to Human PapillomavirusAuthor : Anggraeni R. Yugis, Agus Saputra, Rachmitasari Noviana, Diah Iskandriati, Joko Pamungkas, Irma H. Suparto
Lactoferrin (Lf) has been reported for its multifunctional properties such as antifungal, antibacterial, antiviral, antioxidant and anticancer activities. This protein is an 80 kDa iron-binding protein of the transferrin family. It can be isolated from milk and has been identified from various mammalian secretions. The aims of this study focused on the isolation and purification of Lf from etawa goat colostrum using several methods of chromatography and observing its activities as antiviral human papillomavirus (HPV). Purification was performed by gel filtration chromatography using Sephadex G-75 and G-100, for cation exchange chromatography with carboxymethyl Sephadex C-50, and two-steps chromatography of cation exchange resin carboxymethyl Sephadex C-50 and gel filtration resin Sephadex G-75. All columns were prepared by manual packaging. The native isolated Lf was characterized by SDS-PAGE electrophoresis. Our result showed that the best purified goat lactoferrin (gLf) was by two-steps chromatography with yield of 364 µg/mL and molecular weight 82 kDa. The pure gLf showed anti-HPV effect at concentration of 100 µg/mL after 72 hours incubation by the increased in cycle threshold (Ct) value, from 26 Ct to 36 Ct.
Keywords: goat lactoferrin, purification, ion exchange chromatography, gel filtration chromatography, antihuman papillomavirus