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

Toxins

Jen-Ron Chiang¹  Ching-Sheng Hung²  Chi-Ching Chen³  Liang-Tzung Lin⁴  Sy-Jye Leu⁴  Bor-Yu Tsai⁵  Yi-Yuan Yang⁶  Chia-I Liu⁶  PharaohFellow Mwale⁶  Chi-Hsin Lee⁶  Yu-Ching Lee⁷ 

[1] Center for Research, Diagnostics and Vaccine Development, Centers for Disease Control, Ministry of Health and Welfare, Taipei 11561, Taiwan;Department of Laboratory Medicine, Wan Fang Hospital, Taipei Medical University, Taipei 11696, Taiwan;Department of Pathology and Laboratory Medicine, Landseed Hospital, Taoyuan 32449, Taiwan;Graduate Institute of Medical Sciences, College of Medicine, Taipei Medical University, Taipei 11031, Taiwan;Navi Bio-Therapeutics Inc., Taipei 10351, Taiwan;School of Medical Laboratory Science and Biotechnology, College of Medical Science and Technology, Taipei Medical University, Taipei 11031, Taiwan;The Center of Translational Medicine, Taipei Medical University, Taipei 11031, Taiwan;

关键词: Naja naja atra (NNA);    venom proteins;    IgY antibodies;    phage display technology;    single-chain variable fragment (scFv) antibody;   

DOI  :  10.3390/toxins10100383

来源: DOAJ

【 摘 要 】

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 × 108 and 8.4 × 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.

【 授权许可】

Unknown