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      Rapid development of affinity matured monoclonal antibodies using RIMMS.

      Hybridoma
      Animals, Antibodies, Monoclonal, immunology, Antibody Affinity, Antibody Specificity, B-Lymphocytes, Blotting, Western, Cloning, Molecular, Enzyme-Linked Immunosorbent Assay, Genes, bcl-2, genetics, Haptens, Humans, Hybridomas, Immunization, Mice, Precipitin Tests, Tumor Cells, Cultured

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          Abstract

          Affinity matured murine monoclonal antibody producing cell lines can now be rapidly generated using a novel repetitive, multiple site immunization strategy designated RIMMS. RIMMS capitalizes on rapid hypermutation and affinity maturation events which occur in B cell populations localized within secondary lymphatic tissue early in response to antigenic challenges. A murine myeloma cell line, P3XBcl-2-13, stably transfected with Bcl-2, enhances the outgrowth of hybridomas following somatic fusion with immune lymphocytes isolated from pooled peripheral lymph nodes (PLN) 8-14 days after the initial immunization. Immunizations somatic fusion, screening and isolation of affinity matured IgG secreting monoclonal antibody cell lines occur within a one month time period. By using RIMMS, we have been able to expedite the isolation of affinity matured monoclonal antibodies to numerous antigens, including a drug hapten.

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