Reshaped Human Monoclonal Antibodies for Therapy and Passive Immunization

摘要

The purpose of the project is to apply reshaping technology for the humanizationof monoclonal antibodies specific for junin and vaccinia viruses. The cloning of immunoglobulin variable region heavy (VH) and light (VK) chain genes was achieved by synthesis and amplification of complementary DNA (cDNA) copied from RNA extracted from the mouse hybridoma cells. The CDR (Complementarity Determining Region) sequences were transplanted into human VH and VK genes. The reshaped VH and VK genes were cloned into expression vectors with Human IgG1 or kappa constant regions and transfected into myeloma cells. Reshaped anti-junin and reshaped anti-vaccinia antibodies showed similar or superior binding to virus compared to the progenitor mouse antibody, the reshaped antibodies appeared to be 10-fold less effective in virus neutralization. The reshaped anti-vaccinia antibody showed 10-fold lower binding than the progenitor mouse antibody and was similarly less effective in neutralization. For therapy, these deficits in reshaped antibody binding and neutralization should be offset by the prospective greater tolerance of man to these antibodies.