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Toxins (Basel). 2017 Oct 27;9(11). pii: E347. doi: 10.3390/toxins9110347.

Single Chain Antibody Fragment against Venom from the Snake Daboia russelii formosensis.

Author information

1
Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei 11031, Taiwan. chihsine@msn.com.
2
The Center of Translational Medicine, Taipei Medical University, Taipei 11031, Taiwan. ycl@tmu.edu.tw.
3
Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei 11031, Taiwan. yllee@tmu.edu.tw.
4
Department of Microbiology and Immunology, School of Medicine, College of Medicine, Taipei Medical University, Taipei 11031, Taiwan. yllee@tmu.edu.tw.
5
Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei 11031, Taiwan. cmbsycl@tmu.edu.tw.
6
Department of Microbiology and Immunology, School of Medicine, College of Medicine, Taipei Medical University, Taipei 11031, Taiwan. cmbsycl@tmu.edu.tw.
7
Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei 11031, Taiwan. ltlin@tmu.edu.tw.
8
Department of Microbiology and Immunology, School of Medicine, College of Medicine, Taipei Medical University, Taipei 11031, Taiwan. ltlin@tmu.edu.tw.
9
Department of Pathology and Laboratory Medicine, Landseed Hospital, Taoyuan 32449, Taiwan. chen3971@landseed.com.tw.
10
Center for Research, Diagnostics and Vaccine Development, Centers for Disease Control, Ministry of Health and Welfare, Taipei 11558, Taiwan. jrc@cdc.gov.tw.
11
School of Medical Laboratory Science and Biotechnology, College of Medical Science and Technology, Taipei Medical University, Taipei 11031, Taiwan. pmwale@medcol.mw.
12
Navi Bio-Therapeutics Inc., Taipei 10351, Taiwan. boryutsai@navibio.com.tw.
13
Department of Laboratory Medicine, Wan Fang Hospital, Taipei Medical University, Taipei 11696, Taiwan. oryx@w.tmu.edu.tw.
14
School of Medical Laboratory Science and Biotechnology, College of Medical Science and Technology, Taipei Medical University, Taipei 11031, Taiwan. yangyuan@tmu.edu.tw.
15
Core Laboratory of Antibody Generation and Research, Taipei Medical University, Taipei 11031, Taiwan. yangyuan@tmu.edu.tw.

Abstract

Russell's vipers containing hemotoxic and neurotoxic venom commonly cause snake envenomation. Horse-derived antivenom is a specific antidote, but its production is expensive and has side effects. Developing a cost-effective and more tolerable therapeutic strategy is favorable. In this study, using glutaraldehyde-attenuated Daboia russelii formosensis (DRF) venom proteins to immunize chickens, polyclonal yolk-immunoglobulin (IgY) antibodies were generated and showed a specific binding affinity. Phage display technology was used to generate two antibody libraries of single-chain variable fragments (scFvs) containing 3.4 × 10⁷ and 5.5 × 10⁷ transformants, respectively. Phage-based ELISA indicated that specific clones were enriched after bio-panning. The nucleotide sequences of scFv-expressing clones were analyzed and classified into six groups in the short linker and four groups in the long linker. These scFv antibodies specifically bound to DRF proteins, but not other venom proteins. Mass spectrometric data suggested that these scFv antibodies may recognize phospholipase A2 RV-4 or RV-7. In vivo studies showed that anti-DRF IgY exhibited complete protective effects and mixed scFv antibodies increased the survival rate and time of mice challenged with a lethal dose of DRF proteins. These antibodies can be potentially applied in a rapid diagnostic method or for treatment in the future.

KEYWORDS:

Daboia russelii formosensis (DRF); IgY antibody; phage display technology; single-chain variable fragment (scFv) antibody

PMID:
29076991
PMCID:
PMC5705962
DOI:
10.3390/toxins9110347
[Indexed for MEDLINE]
Free PMC Article

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