Separation of egg yolk immunoglobulins using an automated liquid chromatography system

An automated liquid chromatography system was developed to carry out the separation of an egg yolk immunoglobulin (IgY) using cation exchange media. Industrially separated egg yolk was diluted 10 times with distilled water, the pH adjusted to 5.5, and the water-soluble protein fraction separated from lipoproteins by sedimentation. The supernatant was filtered and then applied to a column packed with a cation exchanger within an automated liquid chromatography system. Different operating conditions were investigated using phosphate buffer in order to assess the effect on recovery and purity. Fractions as pure as 80% could be collected and a recovery of the chromatography step of about 65% was obtained for a purity of 60% using either a linear or step gradient. The overall recovery for the process was 34% if one-step dilution/extraction is used for lipoprotein separation by sedimentation, and 51% if two-step dillution/extraction is used. Further improvement of the yield to about 60% is possible using centrifugation for lipoprotein separation. The automated system confers many advantages, the key elements being the time savings and accurate control of the process. © 1992 John Wiley & Sons, Inc.