Expression of recombinant anti‐breast cancer immunotherapeutic monoclonal antibody in baculovirus–insect cell system
作者： Jeong-Hwan Lee , Kyung-A Hwang , SungSu Park , Young-Kug Choo , Kisung Ko
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摘要: 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, culture flexibility and glycosylation capability. system successfully established for mAb fused with KDEL (Lys–Asp–Glu–Leu) endoplasmic reticulum (ER) retention signal (mAb BR55K). heavy chain (HC) light (LC) genes were cloned under control polyhedrin (PPH) P10 promoters, respectively, pFastBacDual vector. gene-expression cassettes carrying both HC LC transferred into a bacmid Escherichia coli (DH10Bac). transfected Sf9 insect cells to generate baculovirus expressing BR55K. Western blot analysis confirmed BR55K baculovirus-infected cells. Cell direct enzyme linked immunosorbent assay (ELISA) showed that mAbs from lysates or medium bound MCF-7 human breast Both purified using Protein A affinity column. Collectively, these results suggest can be expressed, properly assembled serve as an alternative production.
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