Production of monoclonal antibodies to human IgM for assay of viral IgM antibodies.
Monoclonal antibodies to human IgM were produced by fusing the Sp 2/0-Ag 14 line of mouse myeloma cells with spleen cells from BALB/c mice immunized with purified human IgM. From 6 clones which secreted antibody to human IgM, the one which produced the highest levels of antibody and grew relatively rapidly was selected for expansion and production of immune reagents for viral IgM antibody assays. Mouse ascitic fluids produced with this clone of cells had antibody titers for human IgM of 1 X 10(-10) by indirect enzyme immunofluorescence assay (EIFA). The monoclonal antibodies were found to belong to subclass 1 of murine IgG, and their specificity was shown to be directed against the Fab portion of the mu chain of human IgM. Antibodies from murine ascitic fluid conjugated with horseradish peroxidase were shown to be suitable for assay of measles IgM antibody by indirect immunoperoxidase staining. Antibodies conjugated with alkaline phosphatase could be used in an indirect EIFA for determination of measles IgM antibodies; use of monoclonal conjugates in this system eliminated the nonspecific activity observed in tests utilizing polyclonal anti-mu reagents. Further, the monoclonal antibodies were highly satisfactory for use in a 'capture' system for viral IgM antibody assays. The availability of monoclonal antibodies to human IgM overcomes problems with specificity, consistency and supply which have previously hindered development and standardization of viral IgM antibody assays.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000914:Antibodies, Viral / Q000032:analysis*; D005455:Fluorescent Antibody Technique; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D007075:Immunoglobulin M / Q000032:analysis* / Q000276:immunology; D007148:Immunoglobulin mu-Chains / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D007140:Immunoglobulin Fab Fragments; D007075:Immunoglobulin M; D007148:Immunoglobulin mu-Chains