463924 Domain Library Approach for Designing Therapeutic Small Antibody with High Cytotoxicity for Tumor Cells
We applied each medium fraction to immobilized metal affinity chromatography (IMAC) to purify expressed diabodies. We step-wisely measured the cytotoxicity of IMAC-refined diabodies at the various diabody concentrations by MTS assay. In this assay, all samples were prepared to less than 50 mM imidazole concentration, because imidazole showed the cytotoxicity against cancer cells at more than 75 mM. In the result of step-wise screening, we got 6 kinds high cytotoxic diabodies which damaged at cancer cells at 100 fM concentration
By means of the screening, we found some rules of high cytotoxicity, specific domain order, target molecules, and specific antibodies for highly active diabody. These results imply that high cytotoxic diabodies have universal rules of structure and using antibodies for diabody.