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.
Highlights
Snake envenomation is considered a major medical problem worldwide, in tropical or subtropical countries, such as Taiwan
The results showed that most scFv antibodies recognized Daboia russelii formosensis (DRF) proteins but did not cross-react with other snake venom proteins
To monitor the humoral response in chickens, IgY antibodies were purified from eggs collected from chickens at various stages of immunization for enzyme-linked immunosorbent assay (ELISA)
Summary
Snake envenomation is considered a major medical problem worldwide, in tropical or subtropical countries, such as Taiwan. Russell’s viper (Daboia russelii), a common venomous snake widely distributed in South and Eastern Asia, can be classified into five subspecies based on their characteristics and colorations. These subspecies include Daboia russelii formosensis in Taiwan; Daboia russelii siamensis in Thailand, Myanmar and China; Daboia russelii russelii (previously named Daboia russelii nordicus) in North India; Daboia russelii pulchella in Sri Lanka and South India; and Daboia russelli limitis in Indonesia and Java [3,4,5]. Due to the significant variations in the components of snake venom proteins associated with geographic regions, victims often present various clinical symptoms caused by bites of different subspecies of Russell’s viper [3,6]. In the presence of all of the components, PLA2 with different isoenzymes is considered one of the major lethal components in crude
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