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Characterization of Chicken-Derived Single Chain Antibody Fragments against Venom of Naja Naja Atra

Authors :
Chia I. Liu
Chi Ching Chen
Yu Ching Lee
Ching Sheng Hung
Pharaoh Fellow Mwale
Liang Tzung Lin
Yi Yuan Yang
Chi Hsin Lee
Bor Yu Tsai
Sy Jye Leu
Jen Ron Chiang
Source :
Toxins, Vol 10, Iss 10, p 383 (2018), Toxins, Volume 10, Issue 10
Publication Year :
2018
Publisher :
MDPI AG, 2018.

Abstract

Traditional, horse-derived antivenin is currently the most efficient treatment against snake bites. However, it is costly and has unpredictable side effects. Thus, alternative, cost-effective strategies for producing antivenin are needed. In this study, we immunized hens with inactivated NNA venom proteins from the cobra Naja naja atra (NNA). Purified yolk IgY antibodies showed specific anti-NNA binding activity comparable to that of the equine-derived antivenin. We used phage display technology to generate two antibody libraries containing 9.0 &times<br />108 and 8.4 &times<br />108 clones with a short or long linker, respectively. The phage ELISA indicated that anti-NNA clones displaying single-chain variable fragments (scFv) were significantly enriched after biopanning. The nucleotide sequences of the light and heavy chain genes of 30 monoclonal scFv antibodies were determined and classified into six groups with the short linker and nine groups with the long linker. These scFv clones specifically bound to NNA proteins but not to venom proteins from other snakes. Their binding affinities were further determined by competitive ELISA. Animal model studies showed that anti-NNA IgY antibodies exhibited complete protective effects, while a combination of scFv antibodies raised the survival rates and times of mice challenged with lethal doses of NNA venom proteins.

Details

Language :
English
ISSN :
20726651
Volume :
10
Issue :
10
Database :
OpenAIRE
Journal :
Toxins
Accession number :
edsair.doi.dedup.....0628dcc5e56c0b8476646fd9a5b006d9