Monoclonal antibodies against amyloid fibril protein AA. Production, specificity, and use for immunohistochemical localization and classification of AA-type amyloidosis.
Monoclonal antibodies against amyloid fibril protein AA were produced by cell fusion of murine P3 X 63-Ag8.653 myeloma cells with spleen cells of immunized Balb/c mice. To increase immunogenicity, protein AA was coupled to horseradish peroxidase (HRP) or human high molecular weight kininogen (HMWK). Using micro-ELISA (enzyme-linked immunosorbent essay) seven hybridoma cell lines secreting antibodies that specifically bind to protein AA have been selected and cloned. When applied to formalin-fixed paraffin sections of a variety of different amyloid types using immunoperoxidase methods, five monoclonal antibodies bound specifically and strongly to amyloid only of the AA type. Since a series of different AA-amyloids could be stained, these reagents may be used to routinely diagnose AA-amyloidosis in tissue sections. A monoclonal antibody against HRP has also been produced that has been utilized to develop a monoclonal peroxidase-antiperoxidases (PAP) complex. When three immunoperoxidase methods were compared, the sensitivity of a conventional rat PAP was comparable to the monoclonal PAP complex, but the latter was easier to handle. Both methods were more sensitive than the indirect immunoperoxidase technique.
D000682:Amyloid / Q000276:immunology*; D000686:Amyloidosis / Q000145:classification / Q000175:diagnosis*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D006735:Horseradish Peroxidase / Q000276:immunology; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007114:Immunization; D007120:Immunochemistry; D007124:Immunoenzyme Techniques; D007704:Kininogens / Q000276:immunology; D009101:Multiple Myeloma / Q000276:immunology; D000685:Serum Amyloid A Protein / Q000276:immunology*; D013154:Spleen / Q000276:immunology
D000682:Amyloid; D000911:Antibodies, Monoclonal; D007704:Kininogens; D000685:Serum Amyloid A Protein; D006735:Horseradish Peroxidase