Expression Of The Chimeric Antibody Fab' Against CD44 In E.coli

HI44a(IgG2a) is an anti-CD44 monoclonal antibody. Its binding site is located at the extracellular domain of CD44 molecule, which is close to the binding site of hyaluronate. Through years of research in our lab, we have drawn such conclusions: (1)HI44a effectively induces both differentiation and apoptosis of AML cells. (2) HI44a effectively inhibits the proliferation and induces apoptosis of THP-1 cells. (3) These activities of the anti-CD44 antibody may be associated with its inhibitory effect on oncogene and anti-apoptosis protein expression, and changes in mitochondrial membrane potential. (4) CD44 is a potential target for AML differentiation- and apoptosis-inducing therapy.As the mouse antibodies are restricted in clinical use due to their high immunogenecity, we constructed a chimeric antibody Fab' against CD44 by genetic engineering to minimize the human anti-mouse antibody response. We amplified the genes of the variable region of heavy chain (VH) and light chain (VL) of the antibody against CD44 from the anti-CD44 single chain Fv antibody fragment (scFv) expression vector by PCR method. Then, VH and VL were cloned into vectors pYZH and...