Abstract

Snake venom protein from Deinagkistrodon acutus (DA protein), one of the major venomous species in Taiwan, causes hemorrhagic symptoms that can lead to death. Although horse-derived antivenin is a major treatment, relatively strong and detrimental side effects are seen occasionally. In our study, yolk immunoglobulin (IgY) was purified from eggs, and DA protein was recognized using Western blotting and an enzyme-linked immunosorbent assay (ELISA), similar to therapeutic horse antivenin. The ELISA also indicated that specific IgY antibodies were elicited after the fifth booster, plateaued, and lasted for at least 3 months. To generate monoclonal single-chain variable fragment (scFv) antibodies, we used phage display technology to construct two libraries with short or long linkers, containing 6.24×108 and 5.28×108 transformants, respectively. After four rounds of biopanning, the eluted phage titer increased, and the phage-based ELISA indicated that the specific clones were enriched. Nucleotide sequences of 30 individual clones expressing scFv were analyzed and classified into four groups that all specifically recognized the DA venom protein. Furthermore, based on mass spectrometry, the scFv-bound protein was deduced to be snake venom metalloproteinase proteins. Most importantly, both IgY and mixed scFv inhibited the lethal effect in mice injected with the minimum lethal dosage of the DA protein. We suggest that together, these antibodies could be applied to the development of diagnostic agents or treatments for snakebite envenomation in the future.

Original languageEnglish
Pages (from-to)71-80
Number of pages10
JournalApplied and Environmental Microbiology
Volume82
Issue number1
DOIs
Publication statusPublished - 2016

Fingerprint

venom
Snake Venoms
venoms
snake
snakes
antibody
antibodies
protein
Antibodies
Bacteriophages
Antivenins
Single-Chain Antibodies
bacteriophages
Proteins
proteins
Enzyme-Linked Immunosorbent Assay
enzyme-linked immunosorbent assay
assay
Horses
enzyme

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Food Science
  • Biotechnology
  • Ecology

Cite this

Antibodies against venom of the snake Deinagkistrodon acutus. / Lee, Chi Hsin; Lee, Yu Ching; Liang, Meng Huei; Leu, Sy Jye; Lin, Liang Tzung; Chiang, Jen Ron; Yang, Yi Yuan.

In: Applied and Environmental Microbiology, Vol. 82, No. 1, 2016, p. 71-80.

Research output: Contribution to journalArticle

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