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박세라,신용규,이경진,이정환,David HEDIN,Thera MULVANIA,이승호,고기성 한국곤충학회 2014 Entomological Research Vol.44 No.1
Colorectal cancer is a commonly diagnosed cancer in the world. Monoclonal antibody (mAb) CO17‐1A recognizes the tumor‐associated antigen GA733, a cell surface glycoprotein highly expressed in colorectal carcinoma cell, which is considered to be applicable for diagnosis and therapeutic treatment against colorectal cancer. In addition antibodies are glycoproteins, efficiently recognize and eliminate specific pathogenic and disease antigens. We have currently established baculovirus insect cell expression system to produce anti‐colorectal cancer mAb CO17‐1A. In this study, mAb CO17‐1A was expressed in the transgenic insect cell line SfSWT4, in which glycosylation processing pathway has been humanized. Immunoblot confirmed that mAb CO17‐1A properly expressed in SfSWT4 insect cells. mAb CO17‐1A was purified using protein G affinity column. In addition, MALDI‐TOF verified that the mAb CO17‐1A fused to KDEL, endoplasmic reticulum (ER) retention signal (mAb CO17‐1AK) had high mannose type of glycan structure. Migration assay showed that insect cell‐derived mAb CO17‐1AK (mAbI CO17‐1AK) with high mannose type of glycan structure was effective as mammalian‐derived mAb CO17‐1A (mAbM CO17‐1A) in inhibition of metastasis. Kinetic analysis of antigen‐antibody interaction using Surface Plasmon Resonance (SPR) confirmed that mAbI CO17‐1AK is efficient to interact with antigen GA733 as mAbM CO17‐1A. These results suggest that the insect cell expression system with the SfSWT4 possibly can be used as a useful alternative way to produce full‐size mAb for cancer immunotherapy.
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Expression and function of recombinant anti-colorectal cancer mAb CO17-1A in SfSWT4 insect cells
Se-Ra Park,Kyung Jin Lee,Jeong-Hwan Lee,David Hedin,Thera Mulvania,Seung Ho Lee,Kisung Ko 한국응용곤충학회 2013 한국응용곤충학회 학술대회논문집 Vol.2013 No.10
Colorectal cancer is the third most commonly diagnosed cancer in the world, nearly all patients diagnosed with this cancer die from it. Antibodies are glycoprotein molecules, which can efficiently recognize and eliminate specific pathogenic and disease antigens. Antibody researches for the last several decades have demonstrated the potential of therapeutic antibodies to fight cancer. Monoclonal antibody (mAb) CO17-1A recognizes the tumor-associated antigen GA733-2, a cell surface glycoprotein highly expressed in colorectal carcinoma cell, which is applicable for preventing and curing colorectal cancer. We have currently established baculovirus insect cell expression system to produce anti-colorectal cancer mAb CO17-1A. In this study, mAb CO17-1A was expressed in the transgenic insect cell line SWT4, which has humanized glycosylation processing pathway. Immunoblot confirmed that mAb CO17-1A properly expressed in SWT4. mAb CO17-1A was purified using protein G affinity column. In addition, Maldi-TOF verified that the mAb fused to KDEL, ER retention signal had high mannose type of glycan structure whereas the mAb without KDEL had partially humanized glycan structure. These results suggest that the insect cell expression system with the SWT4 possibly can be used as a useful alternative way to produce full-size mAb with humanized glycan structures for cancer immunotherapy.
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