Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2024 Jun 28:23:100201.
doi: 10.1016/j.toxcx.2024.100201. eCollection 2024 Sep.

Generation of chicken-based IgY polyclonal antibodies against Dendroaspis polylepis and preclinical evaluation of envenomation-neutralizing efficacy vis-à-vis selected commercial antivenoms

Stephen Wilson Kpordze  1   2 , Victor Atunga Mobegi  3 , Gideon Mutie Kikuvi  4 , Joseph Kangangi Gikunju  5 , Courage Kosi Setsoafia Saba  6   2 , Jackan Moshe  5 , James Hungo Kimotho  7
Affiliations

Generation of chicken-based IgY polyclonal antibodies against Dendroaspis polylepis and preclinical evaluation of envenomation-neutralizing efficacy vis-à-vis selected commercial antivenoms

Stephen Wilson Kpordze et al. Toxicon X. .

Abstract

The Black mamba, D. polylepis, is one of the many venomous snakes found in Kenya, and known to account for some snakebite incidents. The Kenyan Ministry of Health data reveals annual 15,000 snakebites occurrences. Also, 1 in 15 people in Kenya gets bitten by a snake, and tragically, 1 in 147 of these individuals die of snakebite yearly. Traditionally, antivenoms for treatment are produced from horse or sheep but have complicated and expensive production issues. Alternative production approaches, such as using IgY antibodies derived from chicken egg yolks, may overcome disadvantages with traditional antivenom manufacturing techniques. In this current study, D. polylepis specific IgY polyclonal antibodies were purified from the egg yolks of chickens immunized with D. polylepis venom. These antibodies were subsequently assessed for their in-vivo neutralizing capacity vis-à-vis commercial antivenoms, PANAF-Premium and VINS. The IgY antibodies were purified by ammonium sulfate precipitation and affinity-chromatography, with quality and specificity determined by SDS-PAGE and ELISA. The LD50 of D. polylepis was found to be 0.54 mg/kg in chicks, and 0.34 mg/kg in mice, respectively. Pool of extracted IgY yielded 2.8 mg/mL concentration. Purified IgY under non-reducing and reducing conditions on SDS-PAGE exhibited a single-protein band of about 183 kDa and two bands (67 kDa and 25 kDa), respectively. The minimum-edematogenic dose was 0.05 μg. Anti-D. polylepis IgY antibodies and two antivenoms demonstrated the capacity to neutralize the toxic activities of D. polylepis venom. This study confirms a successful IgY generation against Black mamba venom for the first time, and observed toxic effects of the venom as well as neutralizing capacity of antivenoms.

Keywords: Anti-D. polylepis IgY antibodies; Antivenom; Neutralizing efficacy; Snake venom.

PubMed Disclaimer

Conflict of interest statement

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Figures

Fig. 1
Fig. 1
Lane 1: IgY under reducing conditions (1A) and lane a, IgY under non-reducing conditions (1B) Lane M: molecular weight marker.
Fig. 2
Fig. 2
Concentrations of the affinity purified antibodies yield per week.
Fig. 3
Fig. 3
Immune response for primary and secondary antibody in serum and egg yolk of immunized Isa-Brown chicken with D. polylepis venom. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)
See this image and copyright information in PMC

References

    1. Aalten M., et al. The clinical course and treatment of black mamba (Dendroaspis polylepis) envenomations: a narrative review. Clin. Toxicol. 2021;59(10):860–868. doi: 10.1080/15563650.2021.1943427. - DOI - PubMed
    1. Abbas A.T., El-Kafrawy S.A., Sohrab S.S., Azhar E.I.A. IgY antibodies for the immunoprophylaxis and therapy of respiratory infections. Hum. Vaccines Immunother. 2019;15(1):264–275. doi: 10.1080/21645515.2018.1514224. - DOI - PMC - PubMed
    1. Ainsworth S., et al. The medical threat of mamba envenoming in sub-Saharan Africa revealed by genus-wide analysis of venom composition, toxicity and antivenomics profiling of available antivenoms. J. Proteonomics. 2018;172:173–189. doi: 10.1016/j.jprot.2017.08.016. - DOI - PubMed
    1. Amro W.A., Al-Qaisi W., Al-Razem F. Production and purification of IgY antibodies from chicken egg yolk. J. Genet. Eng. Biotechnol. 2018;16(1):99–103. doi: 10.1016/j.jgeb.2017.10.003. - DOI - PMC - PubMed
    1. Araújo A.S., Lobato Z.I.P., Chávez-Olórtegui C., Velarde D.T. Brazilian IgY-Bothrops antivenom: studies on the development of a process in chicken egg yolk. Toxicon. 2010;55(4):739–744. doi: 10.1016/j.toxicon.2009.11.004. - DOI - PubMed

LinkOut - more resources