Monoclonal Antibody to 5-Bromo- and 5-Iododeoxyuridine: A New Reagent for Detection of DNA Replication

TLDR

Hybridomas were generated by fusing spleen cells from mice immunized with an iodouridine–ovalbumin conjugate with the SP2/0Ag14 plasmacytoma line. The resulting monoclonal antibodies are highly specific for bromodeoxyuridine and iododeoxyuridine, do not cross‑react with thymidine, and enable rapid detection of DNA synthesis in cultured cells after as little as 6 minutes of bromodeoxyuridine exposure by immunofluorescent staining and flow cytometry.

Abstract

Monoclonal antibodies specific for 5-bromodeoxyuridine have been produced and applied in detecting low levels of DNA replication on a cell-by-cell basis in vitro. The immunoglobulin-producing hybridomas were derived from spleen cells of mice immunized with a conjugate of iodouridine and ovalbumin. The cells were fused with the plasmacytoma line SP2/0Ag14. The antibodies produced are highly specific for bromodeoxyuridine and iododeoxyuridine and do not cross-react with thymidine. DNA synthesis in cultured cells exposed to bromodeoxyuridine for as short a time as 6 minutes can be detected easily and rapidly by an immunofluorescent staining method and quantitated by flow cytometry.