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United States Department of Agriculture

Agricultural Research Service

Title: An Improved Chromatographic Method for the Separation of Egg Yolk Igg into Subpopulations Utilizing Immobilized Metal Ion (Fe+3) Affinity Chromatography

item Greene, Cam
item Holt, Peter

Submitted to: Journal of Immunological Methods
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: September 5, 1997
Publication Date: N/A

Interpretive Summary: An improved technique is described for the separation of yolk antibodies into subpopulations based upon activity and other parameters. This improved methodology has direct industrial application and can be used for testing the efficiency of oil-emulsion and recombinant vaccines by measuring the relative levels of the different antibodies produced following vaccination.

Technical Abstract: An improved methodology is described for the separation of yolk IgG into subpopulations using Immobilized Metal Ion (ferric) Affinity Chromatography. The yolk IgG was first extracted using a pre-chilled, pre-acidified method. After extraction, the yolk IgG was then fractionated using an iron column. Using an ascending pH gradient, four IgG containing peaks were well resolved based upon the elution pH, specific activity and the Avidity Index using 6M urea. This technique also allowed for the differentiation between those birds that were artificially infected with Salmonella enteritidis and those birds that were immunized with an oil-emulsion vaccine.

Last Modified: 4/22/2015
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