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.
Ma
Siyuan
S
Institute of Burn Research, Southwest Hospital, State Key Laboratory of Trauma, Burns and Combined Injury, The Third Military Medical University, Gao Tanyan Main Street 30th, 400038, Shapingba District, Chongqing, People's Republic of China.
Zhang
Yaping
Y
D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000302:isolation & purification*; D002645:Chickens / Q000276:immunology*; D002850:Chromatography, Gel; D004530:Egg Yolk / Q000737:chemistry* / Q000276:immunology; D004531:Eggs / Q000032:analysis; D004926:Escherichia coli / Q000276:immunology; D004927:Escherichia coli Infections / Q000276:immunology* / Q000382:microbiology; D005260:Female; D006801:Humans; D007114:Immunization; D007136:Immunoglobulins / Q000276:immunology / Q000302:isolation & purification*; D007158:Immunologic Techniques; D008070:Lipopolysaccharides / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C
D000906:Antibodies; C016512:IgY; D007136:Immunoglobulins; D008070:Lipopolysaccharides
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