Novel monoclonal antibodies to double stranded DNA that require Ca2+ or Mg2+ for their binding.
Murine monoclonal antibody (MoAb) 2B5 (IgG2aK) was obtained by its binding on a solid phase to double-stranded (ds) DNA from one of the mutant CBA/K1(CBA/K1Jms-1prcg/1prcg) mice which were recently found, in our institute, to develop lymphadenopathy associated with the production of anti-double-stranded (ds) antibodies. MoAb 2B5 was highly specific for dsDNA, as shown by enzyme-linked immunosorbent assay (ELISA). The dsDNA binding of 2B5 was decreased dose-dependently by the chelating agent EDTA, being lost completely with 2.5-5.0 mM EDTA, whereas dsDNA on the solid phase remained intact after incubation with EDTA. Addition of Ca2+ or Mg2+ to antibody in culture supernatant that had lost dsDNA binding activity by dialysis against Ca2+ and Mg(2+)-free buffer restored its binding with dsDNA to the original level, indicating that MoAb 2B5 requires Ca2+ or Mg2+ for its binding with dsDNA. It is unknown whether MoAb 2B5 recognizes new conformational epitopes created in the presence of Ca2+ or Mg2+, but this MoAb should be useful in studies on the modes of interaction of DNA with antibodies and DNA-binding proteins.
Imaoka
K
K
Department of Molecular Oncology, University of Tokyo, Japan.
Kanai
Y
Y
D000818:Animals; D000974:Antibodies, Antinuclear / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D000936:Antigen-Antibody Complex / Q000187:drug effects; D002118:Calcium / Q000494:pharmacology*; D004247:DNA / Q000276:immunology*; D004305:Dose-Response Relationship, Drug; D004492:Edetic Acid / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D006825:Hybridomas; D008274:Magnesium / Q000494:pharmacology*; D051379:Mice
D000974:Antibodies, Antinuclear; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D004247:DNA; D004492:Edetic Acid; D008274:Magnesium; D002118:Calcium