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

• Published in: Toxins 2017-10, Vol.9 (11), p.347
• Main Authors: Lee, Chi-Hsin, Lee, Yu-Ching, Lee, Yueh-Lun, Leu, Sy-Jye, Lin, Liang-Tzung, Chen, Chi-Ching, Chiang, Jen-Ron, Mwale, Pharaoh Fellow, Tsai, Bor-Yu, Hung, Ching-Sheng, Yang, Yi-Yuan
• Format: Article
• Language: English
• Subjects: Animals; Antibodies; Antibodies, Neutralizing - immunology; Antibody libraries; Antivenins - immunology; Antivenom; Chains; Chickens; Chickens - immunology; Daboia; Daboia russelii; Daboia russelii formosensis; Daboia russelii formosensis (DRF); Diagnostic systems; Display devices; Dosage; Enzyme-linked immunosorbent assay; Glutaraldehyde; IgY antibody; Immunization; Immunoglobulins; Immunoglobulins - immunology; In vivo methods and tests; Lethal dose; Neurotoxicity; Panning; Phage display; phage display technology; Phages; Phospholipase; Phospholipase A2; Poultry; Proteins; Reptilian Proteins - immunology; Side effects; Single-Chain Antibodies - immunology; single-chain variable fragment (scFv) antibody; Snakes; Spectrometry; Venom; Viper Venoms - immunology; Yolk
• ISSN: 2072-6651; 2072-6651
• DOI: 10.3390/toxins9110347

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 (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.