Campus Units

Animal Science

Document Type

Article

Publication Version

Published Version

Publication Date

10-1-2014

Journal or Book Title

Poultry Science

Volume

93

Issue

10

First Page

2668

Last Page

2677

DOI

10.3382/ps.2014-04093

Abstract

A study was conducted to develop a simple sequential separation protocol to separate phosvitin and IgY from egg yolk without using organic solvents. Egg yolk was diluted with 2 volumes of distilled water (DW), homogenized, and centrifuged. The precipitant was collected and homogenized with 4 volumes of 10% NaCl (wt/vol) in 0.05 N NaOH solution to extract phosvitin. The pH of the homogenate was adjusted to 4.0 and the precipitate was removed by centrifugation. The supernatant was collected and then heat-treated at 70°C for 30 min and centrifuged to remove impurities. The supernatant containing phosvitin was collected, had salts removed, and was concentrated and then freeze-dried. The supernatant from the centrifugation of diluted egg yolk was diluted again with 3 volumes of DW, and the precipitate was removed by centrifugation. The resulting supernatant was concentrated using ultrafiltration and then IgY was precipitated using 20% saturated (NH4)2 SO4 + 15% NaCl (wt/vol). The precipitant was collected after centrifugation at 3,400 × g for 30 min at 4°C and dissolved with DW, had salts removed, and then was freeze-dried. The purity of separated phosvitin and IgY was checked using SDS-PAGE and the proteins were verified using Western blotting. The purity of phosvitin and IgY was 97.2 and 98.7%, and the yield was 98.7 and 80.9%, respectively. The ELISA results indicated that the activities of separated IgY and phosvitin were 96.3 and 98.3%, respectively. This study proved that both phosvitin and IgY can be separated in sequence from egg yolk without using an organic solvent. Also, the method is very simple and has a high potential for scale-up processing.

Comments

This article is published as Lee, Hyun Yong, E. D. N. S. Abeyrathne, Inwook Choi, Joo Won Suh, and Dong UK Ahn. "Sequential separation of immunoglobulin Y and phosvitin from chicken egg yolk without using organic solvents." Poultry science 93, no. 10 (2014): 2668-2677. doi:10.3382/ps.2014-04093.

Copyright Owner

Poultry Science Association Inc.

Language

en

File Format

application/pdf

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