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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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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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Authors and Affiliations

  1. Department of Systems Immunology, College of Biomedical Science, Kangwon National University, Chuncheon, 24341, Korea

    Jun-Yeol Yoon, Sangkyu Kim, Dain Kim, Gyunghee Jo & Hyo Jeong Hong

  2. Division of diagnostic research and development, ImmuneMed Inc., Chuncheon, 24232, Korea

    Dong-Hwan Kim, Jeongha Yoo, Heui Keun Kang, Min Soo Kim & Young-Jin Kim

  3. Armed Force Chemical, Biological, and Radiological Defense Command, Chuncheon, Korea

    Moon-Sik Shin

  4. Institute for National BioDefense Research, College of Life Science & Biotechnology, Korea University, Seoul, 02841, Korea

    Nam-Taek Lee

  5. Institute of Bioscience and Biotechnology, Kangwon National University, Chuncheon, 24341, Korea

    Hyo Jeong Hong

  6. Department of Microbiology, College of Medicine, Hallym University, Chuncheon, 24252, Korea

    Yoon-Won Kim

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  1. Jun-Yeol Yoon
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  2. Dong-Hwan Kim
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  3. Sangkyu Kim
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  9. Min Soo Kim
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  10. Young-Jin Kim
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  11. Nam-Taek Lee
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  13. Yoon-Won Kim
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Correspondence to Yoon-Won Kim.

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First two author’s contributed equally to this work.

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Yoon, JY., Kim, DH., Kim, S. et al. Generation of a monoclonal antibody that has reduced binding activity to VX-inactivated butyrylcholinesterase (BuChE) compared to BuChE by phage display. Biotechnol Bioproc E 22, 114–119 (2017). https://doi.org/10.1007/s12257-017-0110-7

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  • DOI: https://doi.org/10.1007/s12257-017-0110-7

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