In vitro wound healing: Inhibition activity of insect-derived mAb CO17-1A in human colorectal cancer cell migration

초록

The monoclonal antibody (mAb) CO17-1A specifically binds to the tumor-associated cell surface glycoprotein GA733 in colorectal cancer cells. Thus, mAb CO17-1A has the potential to act as an immune therapeutic protein against colorectal cancer. Recently, it was shown that the baculovirus insect cell expression system produces anti-colorectal cancer mAb CO17-1A. In this study, the colorectal cancer antibody mAb CO17-1A fused to the endoplasmic reticulum (ER) retention signal sequence (KDEL), and the (mAb CO17-1AK) was expressed in Spodoptera frugiperda Sf9 insect cells. The yield, cell cytotoxicity, and in vitro anti-tumor activity of mAb CO17-1AK were verified. Western blotting was performed to confirm that both heavy and light chains of mAb CO17-1A were expressed in Sf9 insect cells. The insect-derived mAb (mAb(I)) CO17-1A was purified using a protein G affinity column. An in vitro wound healing assay was conducted to determine the inhibition activity of mAb CO17-1A during tumor cell migration, showing that mAb(I) CO17-1AK was effective as mammalian-derived mAb CO17-1A (mAb(M) CO17-1A). These results suggest that the insect cell expression system can produce and properly assemble mAbs that inhibit tumor cell migration.

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