Molecular cloning and chimerisation of CDI 315B monoclonal antibody

A chimeric mouse-human antibody has been created that recognizes an antigen found on breast cancer cells and melanoma cells. Immunoglobulin constant d omains of mouse monoclonal antibody CDI 3 15B C gamma 1 and CK, were substi tuted by the human C gamma 1 and C kappa. The CDI 315B variable heavy and l ight chain regions were PCR amplified from hybridoma RNA and sequenced. Mou se variable V-H and V-L regions were joint to human IgG(1) and kappa consta nt regions and subcloned into pcDNA3 expression vectors. The Sp2/0 murine m yeloma cells were transfected with expression vectors pcDNA3L and pcDNA3H a nd the reactivity of chimeric antibodies was tested by indirect ELISA using B16F1 murine melanoma cells as well as MCF7 human breast cancer cells, as antigen.