Generation of a monoclonal antibody that has reduced binding activity to VX-inactivated butyrylcholinesterase (BuChE) compared to BuChE by phage display |
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Authors: | Jun-Yeol Yoon Dong-Hwan Kim Sangkyu Kim Dain Kim Gyunghee Jo Moon-Sik Shin Jeongha Yoo Heui Keun Kang Min Soo Kim Young-Jin Kim Nam-Taek Lee Hyo Jeong Hong Yoon-Won Kim |
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Affiliation: | 1.Department of Systems Immunology, College of Biomedical Science,Kangwon National University,Chuncheon,Korea;2.Division of diagnostic research and development,ImmuneMed Inc.,Chuncheon,Korea;3.Armed Force Chemical, Biological, and Radiological Defense Command,Chuncheon,Korea;4.Institute for National BioDefense Research, College of Life Science & Biotechnology,Korea University,Seoul,Korea;5.Institute of Bioscience and Biotechnology,Kangwon National University,Chuncheon,Korea;6.Department of Microbiology, College of Medicine,Hallym University,Chuncheon,Korea |
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Abstract: | Organophosphate (OP) nerve agents are known as the most toxic chemical warfare agents that act by inhibiting the enzyme acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE). Because BuChE is present at a level of about 3,900 times higher than AChE in plasma, most OP agents first react with BuChE in plasma, suggesting that OP-inactivated BuChE (OP-iBuChE) may act as a biomarker of OP exposure. In this study, we generated an anti-BuChE monoclonal antibody (mAb) that has reduced binding activity to VX-inactivated BuChE compared to native BuChE by phage display. We performed subtractive biopanning of a synthetic human Fab library against native BuChE and soman-iBuChE or VX-iBuChE. As the results, we isolated four Fab clones that showed differential binding activities to VX-iBuChE and native BuChE in ELISAs. To confirm the antigen-binding specificity of the selected clones, the Fabs were converted to IgG1s, and the IgG antibodies were expressed in HEK293F cells and purified. One of them (A2) showed approximately 30% reduced binding activity to VX-iBuChE compared to BuChE in a dose-dependent manner, whereas the other three antibodies showed almost the same binding activities to VX-iBuChE and BuChE. In addition, the A2 antibody did not show reduced binding activity to sarin-iBuChE or soman-iBuChE compared to native BuChE. The results indicate that A2 antibody shows reduced binding activity only to VX-iBuChE. A2 antibody may be applied to specific diagnosis of VX exposure. |
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