Generation and application of chicken egg-yolk antibodies

doi:10.1016/S1095-6433(01)00508-6

Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology

Volume 131, Issue 3, March 2002, Pages 569-574

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https://doi.org/10.1016/S1095-6433(01)00508-6 Get rights and content

Abstract

Despite the fact that the use of chicken as immunization host brings many advantages to the production of polyclonal antibodies, the generation of egg yolk immunoglobulins (IgY) is rarely chosen. In this review, we report on the fast and efficient method for generation and affinity purification of IgY, in this case raised against the α-subunit of hypoxia-inducible factor-1 (HIF-1). The IgY antibody was successfully applied in a variety of methods and a number of different species for HIF-1α detection. In electrophoretic mobility shift assays, the IgY antibody recognized the native HIF-1 complex. The IgY antibody also detected HIF-1α protein on Western blots with extracts derived from human, monkey, pig, dog and mouse cell lines grown under hypoxic conditions. Immunofluorescence and immunoprecipitation experiments using the IgY antibody allowed detection and subcellular localization of HIF-1α in the nuclei of hypoxic cells. Chicken antibody production brings great benefit concerning the welfare of the immunized animals, due to non-invasive antibody harvesting with the added convenience of simple egg collection. An additional advantage is the fast and simple IgY isolation from egg yolk. IgY technology is a great improvement and should be considered as a good alternative to conventional polyclonal antibody production in mammals.

Section snippets

An introduction to chicken egg yolk antibodies

Using chicken as the immunization host brings a number of advantages to antibody production, the most apparent being the non-invasive collection of antibodies. As described more than 100 years ago, avian maternal antibodies are transferred from serum to egg yolk to confer passive immunity to embryos and neonates (Klemperer, 1893). IgY antibody production exploits this by taking the antibodies from the egg yolk, hence the term immunoglobulin of egg yolk (IgY). Another advantage is the enhanced

Adaptation to reduced oxygen supply

Adaptation to reduced oxygen supply, i.e. hypoxia, involves specific activation of oxygen-regulated genes, such as erythropoietin, vascular endothelial growth factor and transferrin (Wenger and Gassmann, 1997). Induction of these genes is mediated by the hypoxia-inducible factor-1 (HIF-1), a heterodimeric transcription factor composed of the two subunits HIF-1α and HIF-1β/ARNT (aryl hydrocarbon receptor nuclear translocator) (Wang et al., 1995), which are both constitutively and ubiquitously

Multiple applications of IgY

Egg yolk antibodies from hens immunized with hHIF-1α_530–825-GST fusion protein were extracted from individual eggs. IgY production was achieved according to the protocol provided in Appendix A. Immunoblot analysis of nuclear fractions from HeLaS3 cells revealed the presence of specific IgY antibodies in eggs collected as early as 1 day after the last antigen injection. Affinity purification of the IgY antibodies was necessary because of strong background signals in these experiments. Based on

Conclusions

In all these experiments, the use of polyclonal IgY antibody was crucial. However, despite its general applicability, IgY technology has scarcely been used to date. It seems therefore of great importance to publicize IgY technology and its inherent advantages. Immunization of hens represents an excellent alternative to generate polyclonal antibodies, because chicken housing is inexpensive, egg collection is non-stressful to hens, isolation and affinity purification of IgY antibodies is fast and

Acknowledgements

The authors wish to thank F. Bootz and M. Rosskopf for their help in generating chicken antibodies. We are also indebted to R.H. Wenger, I. Kvietikova, P. Spielmann and U. Ziegler for their advice, help and critical reading of this manuscript, and C. Gasser for the artwork. This project was supported by the Swiss National Science Foundation (31-56743.99) and the Käthe Zingg-Schwichtenberg Fonds.

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Citation Excerpt :

Egg yolk antibody (IgY) is an antibody against specific bacteria purified from the eggs of sterile laying hens immunized with inactivated bacteria, and it is the most important immunoglobulin in egg yolk with strong specificity (Thirumalai et al., 2019). Its preparation meets the ethical requirements of animal protection and involves low cost, convenient preparation, and convenient mass production (an average hen can lay 300 eggs in a year and approximately 40 g of IgY can be extracted) (Tini et al., 2002). Moreover, the specific IgY produced in chickens comprise 1%–10% of the total amount of antibodies.

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^☆: This paper was originally presented at a symposium on extremophiles held within the ESCPD 21st International Congress, Liège, Belgium, 24–28 July 2000.

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Generation and application of chicken egg-yolk antibodies☆

Abstract

Section snippets

An introduction to chicken egg yolk antibodies

Adaptation to reduced oxygen supply

Multiple applications of IgY

Conclusions

Acknowledgements

Poul. Sci.

J. Biol. Chem.

J. Biol. Chem.

Immunol. Today

Kidney Int.

General applicability of chicken egg yolk antibodies: the performance of IgY immunoglobulins raised against the hypoxia-inducible factor 1alpha

FASEB J.

Peroral immunotherapy with yolk antibodies for the prevention and treatment of enteric infections

Immunol. Res.

Induction and nuclear translocation of hypoxia-inducible factor-1 (HIF-1): heterodimerization with ARNT is not necessary for nuclear accumulation of HIF-1alpha

J. Cell Sci.

A comparative study between the adsorption of IgY and IgG on latex particles

Biomater. Sci. Polym. Ed.

Comparison of standard methods for the preparation of egg yolk antibodies

Tierärztliche Praxis

IgY antiporcine endothelial cell antibodies effectively block human antiporcine xenoantibody binding

Xenotransplantation

Efficient production of chicken egg yolk antibodies against a conserved mammalian protein

FASEB J.

Functional interference between hypoxia and dioxin signal transduction pathways: competition for recruitment of the ARNT transcription factor

Mol. Cell Biol.