Comparison Of Anti-Hepatitis B Surface Antigen Monoclone Purification Technology

Hepatitis B is a worldwide epidemic disease. The diagnosis and prevention of the disease is very significant. Using mouse ascites originated mouse anti-HBsAg antibodies, hepatitis B virus surface antigen colloidal gold reagent can screen HBsAg in human blood.In this paper, hepatitis B virus surface antigen monoclonal antibody purification process was studied and different methods were used to compare the purified effect. With double ammonium sulfate. octanoic acid joint ammonium sulfate method, ammonium sulfate - Protein G affinity chromatography and MEP method, monoclonal antibodies from Hybridoma clones were extracted and purified.Using polyacrylamide gel electrophoresis （SDS-PAGE） and enzyme-linked immunosorbent assay （ELISA） detect the yield, purity and titer of purified antibodies. Purified antibody marked on the colloidal gold to test the sensitivity and specificity of the antibodies.Based on the above experiment, the conclusion was concluded as below. With octanoic acid ammonium sulfate joint method, on the concentration of 3% octanoic acid, the yield of antibodies are to be 1.8%, the purity is 79%, and the titer is 2¹² , which is the best result in these four methods. Thus, octanoic acid ammonium sulfate joint is the most method. It can be suggested to use in large-scale production.