METHOD OF PREPARING SURFACE ANTIGEN OF HEPATITIS B VIRUS (HBS AG), MONOCLONAL ANTIBODIES CB-HEPI, SURFACE ANTIGEN OF HEPATITIS B VIRUS (HBS AG) AND METHOD OF PREPARING HYBRID CLONE 48/1/574 PRODUCING MONOCLONAL ANTIBODIES CB-HEPI

摘要

FIELD: genetic engineering, vaccines against hepatitis B. SUBSTANCE: method of preparing the surface antigen of hepatitis B virus (HBsAg) involves culturing yeast strain Pichia pastoris C226 transformed with recombinant plasmid TAO 906. Cells are analyzed and antigenic fraction is purified successively using monoclonal antibodies CB-HEPI. Antigen is obtained as immunogenic particles (size is 22 nm), molecular mass is 24000 Da of the enhanced homogeneity. Monoclonal antibodies CB-HEPI used for antigen purification by immunoaffinity method are produced by hybrid clone 48/1/574 and belong to isotype Ig G2b or IgM. They recognize continuous epitope on the general antigenic determinant "a" of the surface antigen of all subspecies of hepatitis B virus. For preparing hybrid clone mice of the strain Balb/c are immunized with HBsAg, spleen cells are fused with myeloma cells SP2/O/Ag14, hybrid clones are cultured on nutrient media and required clone producing monoclonal antibodies CB-HEPI is selected. EFFECT: improved method of antigen preparing. 5 cl, 10 dwg, 6 ex