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.