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.

Original languageEnglish
Article number347
JournalToxins
Volume9
Issue number11
DOIs
Publication statusPublished - Nov 1 2017

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Single-Chain Antibodies
Immunoglobulin Fragments
Snake Venoms
Venoms
Bacteriophages
Antibodies
Proteins
Russell's Viper
Clone Cells
Antivenins
Snake Bites
Antidotes
Phospholipases A2
Glutaral
Horses
Libraries
Immunoglobulins
Chickens
Survival Rate
Nucleotides

Keywords

  • Daboia russelii formosensis (DRF)
  • IgY antibody
  • Phage display technology
  • Single-chain variable fragment (scFv) antibody

ASJC Scopus subject areas

  • Toxicology
  • Health, Toxicology and Mutagenesis

Cite this

Single chain antibody fragment against venom from the snake Daboia russelii formosensis. / Lee, Chi Hsin; Lee, Yu Ching; Lee, Yueh Lun; Leu, Sy Jye; Lin, Liang Tzung; Chen, Chi Ching; Chiang, Jen Ron; Fellow, Pharaoh; Tsai, Bor Yu; Hung, Ching Sheng; Yang, Yi Yuan.

In: Toxins, Vol. 9, No. 11, 347, 01.11.2017.

Research output: Contribution to journalArticle

Lee, Chi Hsin ; Lee, Yu Ching ; Lee, Yueh Lun ; Leu, Sy Jye ; Lin, Liang Tzung ; Chen, Chi Ching ; Chiang, Jen Ron ; Fellow, Pharaoh ; Tsai, Bor Yu ; Hung, Ching Sheng ; Yang, Yi Yuan. / Single chain antibody fragment against venom from the snake Daboia russelii formosensis. In: Toxins. 2017 ; Vol. 9, No. 11.
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