Although contribution of light chain to DNA reactivity of some murine anti-DNA antibodies (Abs) has been demonstrated, similar studies on human anti-DNA Abs are limited. To investigate this contribution, we reproduced Fab molecules on the surface of phages from a human B cell line producing IgM anti-DNA monoclonal Ab (NE-1) by an Ab-phage display technique. Expressed Fab molecules (p4-1 clone) were similar to the parental mAb in their binding activities and idiotypic expression. We constructed a light chain shuffled library containing Vkappa genes derived from peripheral blood lymphocytes of a patient with systemic lupus erythematosus (SLE) in combination with the NE-1 heavy chain gene. After panning to ss- or dsDNA, 7 Fab-phage clones which showed significant bindings to ss- or dsDNA were isolated. Many other Fab-phage clones from the library did not bind to ss- nor dsDNA. Sequence analysis revealed that light chains of the 7 clones are derived from diverse Vkappa germline genes including rarely used ones such as the L5 and A30. Most of the Vk germline genes have been used for previously reported anti-DNA antibodies. These findings suggest that diverse Vkappa genes can pair with the NE-1 heavy chain for anti-DNA Ab activity. In addition, kappa light chains seem to modulate DNA binding activities in various ways.