Purification of anti‐colorectal cancer monoclonal antibody CO17‐1A from insect cell culture using a French press and sonication

摘要: Monoclonal antibody (mAb) CO17-1A binds to GA733, which is a tumor-associated glycoprotein antigen highly expressed on the colorectal cancer cell surface. Thus, mAb considered useful biomolecule for diagnosis and treatment against cancer. Previously, we established baculovirus–insect expression system production of CO17-1A. In order use as diagnostic therapeutic tool, however, must be properly purified from insect cells. this study, our aim was investigate effective purification processes in Spodoptera frugiperda (Sf9) cells, using French press sonication disruption. SDS-PAGE confirmed that both fused KDEL endoplasmic reticulum (ER) retention signal (mAb CO17-1AK) were clearly Sf9 Western blot analysis showed detection levels CO17-1AK higher when cells disrupted two times by then sonicated, compared only one disruption plus sonication. Optical microscopy treated with disrupted. Analysis gene sequence information verified peptide present but transmembrane protein does not exist. These results suggest twice once an method improving efficiency.