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KMID : 1094720170220020114
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Biotechnology and Bioprocess Engineering
2017 Volume.22 No. 2 p.114 ~ p.119
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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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Yoon Jun-Yeol
Kim Dong-Hwan
Kim Sang-Kyu
Kim Da-In
Jo Gyung-Hee
Shin Moon-Sik
Yoo Jeong-Ha
Kang Heui-Keun
Kim Min-Soo
Kim Young-Jin
Lee Nam-Taek
Hong Hyo-Jeong
Kim Yoon-Won
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Abstract
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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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KEYWORD
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butyrylcholinesterase, diagnosis, monoclonal antibody, nerve toxic agents, organophosphate, phage display
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