Anti-Trimeresurus albolabris venom IgY antibodies: preparation, purification and neutralization efficacy

 

Hai-long Duan¹, Qi-yi He¹, Bin Zhou², Wen-wen Wang¹, Bo Li¹, Ying-zheng Zhang¹, Qiu-ping Deng¹, Ying-feng Zhang¹, Xiao-dong Yu¹

 

1 Animal Toxin Group, Chongqing Key Laboratory of Animal Biology, Chongqing Engineering Research Center of Bioactive Substance, Engineering Research Center of Active Substance and Biotechnology, Ministry of Education, College of Life Science, Chongqing Normal University, Chongqing 401331, China.

2 Library, Chongqing Normal University, Chongqing 401331, China.

 

ABSTRACT

 

Background

Snakebite incidence in southwestern China is mainly attributed to one of the several venomous snakes found in the country, the white-lipped green pit viper Trimeresurus albolabris. Since antivenom produced from horses may cause numerous clinical side effects, the present study was conducted aiming to develop an alternative antivenom antibody (immunoglobulin Y - IgY) from leghorn chickens.

 

Methods

IgY in egg yolk from white leghorn chicken previously injected with T. albolabris venom was extracted by water, precipitated by ammonium sulfate and purified by affinity chromatographic system. IgY was identified by SDS-PAGE, ELISA and Western blot, and its neutralizing assay was conducted on mice.

 

Results

Chickens injected multiple times with T. albolabris venom elicited strong antibody responses, and from their egg yolk IgY was isolated and purified, which exhibited a single protein band on SDS-PAGE and two bands (about 65 and 35 kDa, respectively) under reduced conditions. Immunoblot analysis revealed that these IgY are polyclonal antibodies since they bind with most venom components. In the neutralizing assay, all mice survived while the ratios of IgY/venom reached up to 3.79 (50.0 mg/13.2 mg).

 

Conclusions

IgY antibody response was successfully conducted in white leghorn chicken injected with T. albolabrisvenom. IgY against T. albolabris venom was obtained for the first time, and it exhibited strong neutralizing potency on mice. These results may lay a foundation for the development of IgY antivenom with clinical applications in the future.

 

Key words: IgY; Egg yolk; Snake venom; Trimeresurus albolabris; LD50

 

Ethics approval and consent to participate

All the experiments were performed according to the guidelines approved by Chongqing Municipal Public Health Bureau.

 

Received: April 1, 2016.

Revised: July 27, 2016.

Accepted: August 24, 2016.

 

Correspondence: yxd@cqnu.edu.cn

 

Authors’ contributions

H-LD and Q-YH equally and mainly contributed to this study. X-DY provided the idea, gave supervision for experimental designs and finalized the manuscript. The other authors helped in all the experiments during the study period. All authors read and approved the manuscript.

 

Competing interests

The authors declare that they have no competing interests.

doi: 10.1186/s40409-016-0078-3