Establishment of human monoclonal anti-DNA antibody producing cell lines

We developed a useful method for the establishment of stable cell lines producing human monoclonal anti-DNA antibody by in vitro Epstein-Barr virus infection. The practical limitation for the cloning was overcome by 2 procedures. One was a microculture system using a small number of the culture. Another was enrichment of anti-DNA producing cells at an early stage and prior to the cloning. The combination of these procedures allowed ready derivation of the cell lines secreting monoclonal anti-DNA antibody. Sixteen cell lines were cloned by utilizing colony formation methods in soft agarose. About 14-32 micrograms per ml of IgM with specific antibody activity were obtained in the supernatant of the cells. The antibody reacted with double-stranded and/or single-stranded DNA. These cells have been continuously producing the specific antibody for more than 3 years. We may extend this procedure for obtaining other autoantibodies, such as anti-T cell antibodies.