Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology
Generation and application of chicken egg-yolk antibodies☆
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-1α530–825-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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This paper was originally presented at a symposium on extremophiles held within the ESCPD 21st International Congress, Liège, Belgium, 24–28 July 2000.