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 × 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.

Original languageEnglish
Article number383
JournalToxins
Volume10
Issue number10
DOIs
Publication statusPublished - Oct 1 2018

Fingerprint

Cobra Venoms
Elapidae
Single-Chain Antibodies
Immunoglobulin Fragments
Venoms
Antivenins
Chickens
Bacteriophages
Antibodies
Proteins
Monoclonal antibodies
Clone Cells
Horses
Animals
Nucleotides
Genes
Enzyme-Linked Immunosorbent Assay
Display devices
Snake Bites
Snakes

Keywords

  • IgY antibodies
  • Naja naja atra (NNA)
  • Phage display technology
  • Single-chain variable fragment (scFv) antibody
  • Venom proteins

ASJC Scopus subject areas

  • Toxicology
  • Health, Toxicology and Mutagenesis

Cite this

Characterization of chicken-derived single chain antibody fragments against venom of Naja naja atra. / Lee, Chi Hsin; Leu, Sy Jye; Lee, Yu Ching; Liu, Chia I.; Lin, Liang Tzung; Mwale, Pharaoh Fellow; Chiang, Jen Ron; Tsai, Bor Yu; Chen, Chi Ching; Hung, Ching Sheng; Yang, Yi Yuan.

In: Toxins, Vol. 10, No. 10, 383, 01.10.2018.

Research output: Contribution to journalArticle

@article{0c6fd1ab35b54da9a515b302a6d66455,
title = "Characterization of chicken-derived single chain antibody fragments against venom of Naja naja atra",
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 × 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.",
keywords = "IgY antibodies, Naja naja atra (NNA), Phage display technology, Single-chain variable fragment (scFv) antibody, Venom proteins",
author = "Lee, {Chi Hsin} and Leu, {Sy Jye} and Lee, {Yu Ching} and Liu, {Chia I.} and Lin, {Liang Tzung} and Mwale, {Pharaoh Fellow} and Chiang, {Jen Ron} and Tsai, {Bor Yu} and Chen, {Chi Ching} and Hung, {Ching Sheng} and Yang, {Yi Yuan}",
year = "2018",
month = "10",
day = "1",
doi = "10.3390/toxins10100383",
language = "English",
volume = "10",
journal = "Toxins",
issn = "2072-6651",
publisher = "Multidisciplinary Digital Publishing Institute (MDPI)",
number = "10",

}

TY - JOUR

T1 - Characterization of chicken-derived single chain antibody fragments against venom of Naja naja atra

AU - Lee, Chi Hsin

AU - Leu, Sy Jye

AU - Lee, Yu Ching

AU - Liu, Chia I.

AU - Lin, Liang Tzung

AU - Mwale, Pharaoh Fellow

AU - Chiang, Jen Ron

AU - Tsai, Bor Yu

AU - Chen, Chi Ching

AU - Hung, Ching Sheng

AU - Yang, Yi Yuan

PY - 2018/10/1

Y1 - 2018/10/1

N2 - 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.

AB - 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.

KW - IgY antibodies

KW - Naja naja atra (NNA)

KW - Phage display technology

KW - Single-chain variable fragment (scFv) antibody

KW - Venom proteins

UR - http://www.scopus.com/inward/record.url?scp=85053816524&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85053816524&partnerID=8YFLogxK

U2 - 10.3390/toxins10100383

DO - 10.3390/toxins10100383

M3 - Article

AN - SCOPUS:85053816524

VL - 10

JO - Toxins

JF - Toxins

SN - 2072-6651

IS - 10

M1 - 383

ER -