Optimized method for the extraction of contaminant-free IgY antibodies from egg yolk using PEG 6000

Hens are oviparous vertebrates and produce IgY antibodies, which is the main type of immunoglobulin in the egg yolk, and high concentrations can be obtained by using a simple method that does not require sophisticated equipment and reagents. The Polyethylene Glycol 6000 method allows the removal of lipids and the precipitation of IgY in two days with an approximated purity of around 80%, however during the original protocol other contaminant proteins can be precipitated. To overcome the issue of contamination with other proteins and extraction time, we optimized the previously method described by Pauly et al. (2011) by adding some changes that improved the aforementioned problems. • Our protocol is customized by the addition of one more filtration step or one more step with PEG 6000 at 3.5% to avoid the contamination with lipids. • Additionally, the changes in the type of agitation, centrifugation and the skip of dialysis make the method more accessible for all the laboratories. • In summary, these modifications serve to enhance the purity, reduce the time for IgY extraction from egg yolk and make it more accessible for every basic research laboratory.