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Monoclonal antibodies to a nuclear protein (PCNA/cyclin) associated with DNA replication

1987; Elsevier BV; Volume: 168; Issue: 2 Linguagem: Inglês

10.1016/0014-4827(87)90020-6

1090-2422

Kenji Ogata, Pekka Kurki, Julio E. Celis, Robert M. Nakamura, Eng M. Tan,

T-cell and B-cell Immunology

Two hybridomas producing monoclonal antibodies to proliferating cell nuclear antigen. (PNCA)/cyclin were generated from spleen cells of BALB/c mice immunized with purified PCNA from rabbit thymus. The specificity of the monoclonal antibodies (19A2 and 19F4) was established by showing that they reacted in enzyme-linked immunosorbent assay (ELISA) with purified PCNA. Furthermore, they reacted in one-dimensional (1D) gel immunoblots with a 36 kD polypeptide which also reacted with human autoantibodies from lupus patients. Both monoclonals also recognized the nuclear polypeptide cyclin in twodimensional (2D) gel immunoblots of HeLa cell proteins. Epitopes recognized by 19A2 and 19F4 were analysed by competitive inhibition test using a modified ELISA. The data suggested that the epitopes were closely related, but not identical. The data with human auto-antibodies were more difficult to interpret, although it suggested that some human anti-PCNA may share epitopes with 19A2 and 19F4, but in addition recognize different epitopes on the PCNA molecule.