Comparison of four purification methods for the production of immunoglobulins from eggs laid by hens immunized with an enterotoxigenic E. coli strain.
The importance of eggs as a source of specific antibodies is well recognized. Egg yolk contains 8-20 mg of immunoglobulins (IgY) per ml. However, the major problem in isolation is removal of lipids which are present in high concentrations. A method had been developed by employing water dilution to separate the yolk plasma proteins from the granules and lipids. Further purification of IgY from plasma proteins was achieved by a protocol involving salt precipitation and ultrafiltration. The water dilution method (WD) was compared with three other methods, namely, polyethylene glycol (PEG), dextran sulphate (DS) and xanthan gum (Xan) in terms of yield, purity, ease of use, potential scaling up and immunoactivity of IgY. The WD method gave the highest yield, followed by DS, Xan and PEG methods in that order. 9.8 mg IgY/ml egg yolk was routinely obtained from the WD method compared to 4.9 mg IgY/ml egg yolk with the popular PEG method with purities of 94% and 89% respectively. Purification methods had no adverse effect on the immunoactivities of IgY. WD was also found superior in terms of ease of use and large scale production of IgY. WD method therefore provides a simple, rapid and efficient means of purifying IgY with high activity.
Akita
E M
EM
Department of Food Science, University of British Columbia, Vancouver, Canada.
Nakai
S
S
D000818:Animals; D000907:Antibodies, Bacterial / Q000302:isolation & purification*; D002645:Chickens; D004527:Egg Proteins / Q000302:isolation & purification*; D004530:Egg Yolk / Q000737:chemistry; D004768:Enterotoxins / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000276:immunology*; D005260:Female; D007136:Immunoglobulins / Q000302:isolation & purification*; D012995:Solubility
D000907:Antibodies, Bacterial; D004527:Egg Proteins; D004768:Enterotoxins; C016512:IgY; D007136:Immunoglobulins