Department of Chemistry & Biomolecular Sciences, Macquarie University, NSW, 2109, Australia.
Chicken IgY antibodies have been touted to be a superior alternative to mammalian antibodies for use in various immunological, molecular biology and proteomics applications for several reasons. These include, but are not limited to, improved specificity due to maximum phylogenetic distance between host and recipient, cost effectiveness in maintaining commercial numbers of hens, IgY yield and the use of non-invasive methods used to isolate IgY from eggs as opposed to blood. Despite this, the routine use of IgY-based methodologies in the laboratory is not widespread. One reason for this reluctance may be derived from the difficulties and expense of isolating IgY antibodies from egg yolk in sufficient yield, with high purity at a realistic reasonable price. Here, we describe an extremely cost-effective ($5USD per egg), rapid (within 5 h), efficient and optimised technique to isolate high yields (60 mg) of high purity (~80%) chicken IgY from egg yolks using the common plant gums pectin and κ-carrageenan in the presence of calcium chloride to delipidate egg yolk mixtures whilst maintaining IgY in solution and then ammonium sulphate to subsequently precipitate the resulting IgY antibodies to higher purity. Our data demonstrates that this technique results in a high yield and purity of IgY that is comparable (if not superior to) existing commercial IgY isolation kits. The method also allows the isolation of immunologically active IgY which can be used for further downstream immunotechnological processes. Furthermore, it can also be easily implemented in a standard well equipped laboratory, and may be scaled up to commercial quantities (i.e., thousands of eggs).
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