The Protein Journal

, Volume 29, Issue 7, pp 475–480 | Cite as

Preparation of Immunoglobulin Y (IgY) Against Lipopolysaccharide using Gel Chromatography from the Yolks of Eggs Laid by Immunized Hens



The objective is to prevent and treat injuries caused by lipopolysaccharide (LPS) from gram negative bacteria in animals and humans, we produced antibodies against LPS from egg yolk. LPS from E. coli (O111:B4) mixed with Freund’s Adjuvant was used as the immunogen to immunize Roman hens. Immunized eggs were collected, and immunoglobulin Y (IgY) was purified using a water solution, salt precipitation and gel chromatography. The molecular weight and purity were determined by SDS–PAGE, the antibody titer by noncompetitive enzyme-linked immunosorbent assay (ELISA) and antibody activity against LPS by the mortality of mice intraperitoneally injected with LPS or LPS-IgY solutions. IgY against LPS showed two protein bands at 68 and 26 kDa on the gel; the antibody titer was almost 1:25,600. After incubation with LPS, IgY decreased the mortality of mice challenged with LPS. This study provided an efficient way to produce high-titer egg yolk antibodies, which could attenuate lethal effects of LPS, by immunizing hens. Furthermore, the LPS antibody was purified well using a water solution, salting-out and gel chromatography.


Purification Lipopolysaccharide Immunoglobulins Egg yolk Antibody 



Immunoglobulin Y; egg yolk antibody



E. coli

Escherichia coli


Hydrogen chloride


Sodium sulfate


Optical density


Phosphate-buffered saline



The authors thank the staff of the State Key Laboratory of Trauma, Burns and Combined Injury, Southwest Hospital, Third Military Medical University, for their assistance in caring for the animals, collecting eggs and providing technical services used in this study.


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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.Institute of Burn Research, Southwest Hospital, State Key Laboratory of Trauma, Burns and Combined InjuryThe Third Military Medical UniversityShapingba District, ChongqingPeople’s Republic of China

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