Expression of recombinant anti-breast cancer immunotherapeutic monoclonal antibody in baculovirus-insect cell system

초록

The anti-breast cancer monoclonal antibody (mAb) BR55 was expressed in the baculovirus-insect cell expression system, which is advantageous because of its high production capacity, cell culture flexibility and glycosylation capability. The baculovirus-insect cell expression system was successfully established for production of mAb BR55 and mAb BR55 fused with the KDEL (Lys-Asp-Glu-Leu) endoplasmic reticulum (ER) retention signal (mAb BR55K). The heavy chain (HC) and light chain (LC) genes of mAb BR55 were cloned under the control of the polyhedrin (P-PH) and P-10 promoters, respectively, in the pFastBacDual vector. The antibody gene-expression cassettes carrying both the HC and LC genes were transferred into a bacmid in Escherichia coli (DH10Bac). The bacmid carrying the expression cassettes was transfected into Sf9 insect cells to generate baculovirus expressing mAb BR55 and BR55K. Western blot analysis confirmed the expression of mAb BR55 and BR55K in baculovirus-infected insect cells. Cell direct enzyme linked immunosorbent assay (ELISA) showed that both mAbs from insect cell lysates or cell culture medium bound to MCF-7 human breast cancer cells. Both mAb BR55 and BR55K were successfully purified using a Protein A affinity column. Collectively, these results suggest that the anti-breast cancer mAb BR55 can be expressed, properly assembled and purified from the baculovirus expression system, which can serve as an alternative system for antibody production.

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