中国生物工程杂志

Objective: Preparation of monoclonal antibody to detect the wild-type HBsAg as well as variant forms. Methods: BALB/c female Mice were immunized with HBsAg isolated from serum of infected patients. mAbs were prepared by hybridoma technology. Identify the purity and characteristics of the selected mAb purified with saturated ammonium sulfate and give preliminary evaluation of the quality and effectiveness of its application, in which polyacrylamide gel electrophoresis and agarose gel electrophoresis and emzyme-linked immunosorbent assay (ELISA) were used. Results: A monoclonal antibody can detect the wild-type HBsAg as well as variant forms was obtained designated as D12. The purity of D12 was high. D12 was highly sensitive and specific to HBsAg and was tested for subtype as IgGl . The recognition sites existed in the nature antigen. The ability to detect the wild-type HBsAg using D12 was better than that of three commercial HBsAg diagnostic kits and the ability to detect the variant HBsAg using D12 was better than that of five commercial HBsAg diagnostic kits. Conclusion: The preparation of mAb D12 is very important to further improve the detection rate of variant hepatitis B virus and enhance HBV prevention.