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Sequential separation of immunoglobulin Y and phosvitin from chicken egg yolk without using organic solvents.

Posted by on in 2014
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  • Lee HY1Abeyrathne ED2Choi I3Suh JW4Ahn DU5. 2014. Poult Sci. 93(10):2668-77. doi: 10.3382/ps.2014-04093. Epub 2014 Aug 1.
    1
    WCU Biomodulation, Department of Agricultural Biotechnology, Seoul National University, Seoul 151-921, South Korea Center for Nutraceutical and Pharmaceutical Materials, Myongji University, Cheoin-gu, Yongin, Gyeonggi-Do 449-728, Korea.
  • 2WCU Biomodulation, Department of Agricultural Biotechnology, Seoul National University, Seoul 151-921, South Korea Department of Animal Science, Uva Wellassa University, Badulla, Sri Lanka 90000.
  • 3Functional Materials Research Group, Food Research Institute, Sungnam, Gyeonggi-Do, 463-746, Korea.
  • 4Center for Nutraceutical and Pharmaceutical Materials, Myongji University, Cheoin-gu, Yongin, Gyeonggi-Do 449-728, Korea duahn@iastate.edu jwsuh@mju.ac.kr.
  • 5WCU Biomodulation, Department of Agricultural Biotechnology, Seoul National University, Seoul 151-921, South Korea Department of Animal Science, Iowa State University, Ames 50011 Department of Animal Science and Technology, Sunchon National University, Sunchon 540-742, South Korea duahn@iastate.edu jwsuh@mju.ac.kr.

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)2SO4 + 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.

©2014 Poultry Science Association Inc.

KEYWORDS:

egg yolk; immunoglobulin Y; phosvitin; sequential separation

PMID:
 
25085938
 
[PubMed - in process]
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