Paper Title
An Integrated Purification System Approached For Polyclonal Anti- Hepatitis B Core Antigen Immunoglobulin G Recovery

Abstract
Immunoglobulin G (IgG) is produced by the plasma cells of animals� immune systems to identify and neutralize pathogens such as bacteria and viruses. In this study, immunoglobulins G (IgG) against hepatitis B core antigen (HBcAg) was purified effectively using an integrated purification scheme comprising of ammonium sulfate precipitation and SepFastTMMM AH-1 column chromatography. Ammonium sulfate precipitation performed at 40% saturation was optimal with respect to the concentration of polyclonal IgG recovered (7.8mg/ml) and the removal of albumin (72%). The yield, purity and purification factor achieved from this simple purification method were 99%, 94% and 7.8, respectively. IgG recovered from ammonium sulfate precipitation was flowed through the SepFastTM MM AH-1 column chromatography and the purity of IgG was further increased to 98%, corresponding to a purification factor of 8.1. Protein aggregation was also reduced significantly in the purified IgG sample. Dynamic light scattering analysis revealed the molecular size of the purified IgG corresponded well to that of the native IgG which was about 10.3 nm. Furthermore, the salt content in the purified sample was reduced by 75% and therefore the need of desalting the final product was eliminated. Enzyme-linked immunosorbent assay (ELISA) showed that the purified anti-HBcAg IgG was highly antigenic. Keywords- Purification; immunoglobulin G; hepatitis B core antigen;ammonium sulphateprecipitation; mixed-mode chromatography.