Generation and characterization of three murine monoclonal nucleotide binding anti-ssDNA autoantibodies.
Three murine monoclonal anti-ssDNA autoantibodies have been produced by fusion of spleen cells from NZB/NZW F1 hybrid mice with Sp 2/0-Ag 14 murine myeloma cells. Specificity studies characterizing monoclonal anti-ssDNA autoantibodies revealed binding patterns for nucleotides. Clone E-4-1 showed preferential binding with mono-, di- and polyguanosine. Clones E-4-4 and E-11-1 specifically bound mono-, di- and polythymidine. Results of inhibition studies indicated an order of reactivity for each monoclonal autoantibody: polynucleotide greater than dinucleotide greater than mononucleotide. In a fluorescence immune assay, using aminonaphthyl derivatives of nucleotides, all three monoclonal antibodies exhibited a low affinity (5 x 10(4)-10(5) M-1) for the homologous mononucleotides. The affinity of clone E-4-1 increased approximately 10-fold when a dinucleotide derivative was used relative to a mononucleotide fluorescent ligand.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D000915:Antibody Affinity; D000918:Antibody Specificity; D001323:Autoantibodies / Q000378:metabolism*; D001667:Binding, Competitive; D004247:DNA / Q000276:immunology; D004277:DNA, Single-Stranded / Q000276:immunology*; D005455:Fluorescent Antibody Technique; D051379:Mice; D008807:Mice, Inbred BALB C; D009711:Nucleotides / Q000276:immunology
D000911:Antibodies, Monoclonal; D001323:Autoantibodies; D004277:DNA, Single-Stranded; D009711:Nucleotides; D004247:DNA