Construction and initial characterization of a mouse-human chimeric anti-TNF antibody

• DOI: 10.1016/0161-5890(93)90106-l
• Publication Date: 2003-03-15T06:59:29Z
• Authors: David M. Knight; Han Trinh; Junming Le; Scott Siegel; David Shealy; Margaret McDonough; Bernard Scallon; Maria Arevalo Moore; Jan Vilcek; Peter Daddona; John Ghrayeb
• Keywords: 0301 basic medicine; Hybridomas; Chimera; Tumor Necrosis Factor-alpha; Antibodies, Monoclonal; Transfection; Recombinant Proteins; 3. Good health; Mice; 03 medical and health sciences; Neutralization Tests; Animals; Humans; Immunoglobulin Light Chains; DNA Probes; Immunoglobulin Heavy Chains

Tumor necrosis factor-alpha (TNF) has been implicated in the pathogenesis of a variety of human diseases including septic shock, cachexia, graft-versus-host disease and several autoimmune diseases. Monoclonal antibodies directed against TNF provide an attractive mode of therapeutic intervention in these diseases. We have generated a murine monoclonal antibody (A2) with high affinity and specificity for recombinant and natural human TNF. To increase its therapeutic usefulness, we used genetic engineering techniques to replace the murine constant regions with human counterparts while retaining the murine antigen binding regions. The resulting mouse-human chimeric antibody should have reduced immunogenicity and improved pharmacokinetics in humans. Molecular analysis of light chain genomic clones derived from the murine hybridoma suggests that two different alleles of the same variable region gene have rearranged independently and coexist in the same hybridoma cell. The chimeric A2 antibody (cA2) exhibits better binding and neutralizing characteristics than the murine A2 which was shown to contain a mixture of two kappa light chains. The properties of cA2 suggest that it will have advantages over existing murine anti-TNF antibodies for clinical use.