• Korean Chemical Engineering Research
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• v.50 no.4
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• pp.659-665
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• 2012

IgY (Immunoglobulin Yolk) in egg yolk corresponds to IgG (Immunoglobulin G) in animal serum and plays an important role as immunological proteins in intestines. Carrageenan and Arabic gum were used as pretreatment agents to purify IgY from fresh egg yolk. DEAE (Diethylaminoethyl) Sepharose column in FPLC (Fast Protein Liquid chromatography) was an ion exchange tool to remove contaminants as well as to elute IgY from the column. GF HPLC (Gel Filtration High Performance Liquid Chromatography) enables to measure the molecular weights of IgY and to identify the purified IgY by comparing the molecular weight of standard IgY with the purified one. IgY is a heterogeneous group of different molecular weight and ionic properties, which was investigated with various IE HPLC (Ion Exchange High Performance Liquid Chromatography) columns such as AX, CX and SCX. Three peaks of IgY were separated in the AX column under the conditions of 0.5 M NaCl and pH=8. The SCX column also gave the three peaks of IgY at 0.5 M NaCl and pH=5.

• KSBB Journal
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• v.14 no.6
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• pp.677-681
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• 1999

Purificationi of egg yolk immunoglobulin(IgY) was performed to understand the property of egg immunoglobulin. IgY differs from mammalian IgY in the molecular size(larger), isoelectric point(more acidic), and binding ability with mammalian complement and protein A(nonbinding ability). IgY is also known as ${\gamma}$-livetin and exists in egg yolk together with other two water-solubel proteins, ${\alpha}$-livetin(chicken serum albumin) and ${\beta}$-livetin(${\alpha}_2$-glycoprotein) and various lipoproteins(Low density lipoprotein, LDL and High density lipoprotein, HDL) which are the major components of egg yolk. The first step of isolation of IgY is to separate the water-solube proteins from lipoproteins. We report a simple method for separation of water soluble proteins using k-carrageenan and sedimentation. k-carrageenan was found to be effective for removal of yolk lipoprotein as a precipitate. IgY remained supernatant, and was isolated by chromatography on columns of DEAE-Sephacel and G 75 gel filtration chromatography.

• Food Science and Biotechnology
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• v.15 no.2
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• pp.189-195
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• 2006

A study was conducted to develop optimal conditions for a large-scale, sequential separation method for value-added components from egg yolk. Starting with liquid egg yolk, immunoglobulin Y (IgY), phospholipids, and neutral lipids were extracted sequentially using water, ethanol, and n-hexane. The remainder was yolk proteins. Adjusting the pH of diluted egg yolk to pH 5.0-5.2 decreased phospholipids content in the supernatant after centrifugation, which was very important for preventing clogging problem of ultrafiltration filters during the subsequent concentration step for IgY separation. Extraction of precipitants after centrifugation with four volumes of 100% ethanol once removed most of the phospholipids and the purity of phospholipids was more than 85% (wt.) after drying. The subsequent extraction of precipitant from ethanol extraction with four volumes of hexane 3 times removed neutral lipids almost completely and resulted in a high protein product with minimal lipids. The sequential separation method is considered to be advantageous for large-scale separations of many valuable components from egg yolk.

• Korean Chemical Engineering Research
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• v.49 no.6
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• pp.798-803
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• 2011

IgY(Immunoglobulin Yolk) is a specific antibody in egg yolk, and it protects human body from virus and antigen. There are a lot of egg yolk components such as lipoprotein and protein. To separate IgY, HPLC(High Performance Liquid Chromatography) and precipitation were used in a batch mode and SMB(Simulated Moving Bed) was adopted for continuous purification of yolk proteins. IgY and other proteins in yolk were separated by using three-zone SMB chromatography. Before performing SMB experiments, batch chromatography and PIM(pulse input method) were performed to find operation parameters and adsorption isotherms. The results of batch chromatography were compared with simulated results using Aspen chromatography. To find the most suitable separation condition in SMB chromatography, simulations in $m_2$-$m_3$ plane on the triangle theory were carried out. $m_2$ = 0.18, $m_3$ = 1.0 and ${\Delta}$t = 419 s are the best conditions for the highest purity of IgY. With this operating parameters(flow rate in three zone and switching time), the purity of raffinate results in 98.39% from Aspen chromatography simulation. Most of the simulation reached steadystate within second recycle.

• Research in Plant Disease
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• v.12 no.2
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• pp.144-147
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• 2006

Egg yolk immunoglobulin (IgY) is much widely used in medical fields, but its use in serology of plant viruses is much limited. We produced an IgY against pepper mild mottle virus (PMMoV) and applied it to several serological tests. Polyclonal antibodies were obtained from the egg yolk of chicken immunized with a total of 2mg of purified PMMoV over 2 months. The titers of antibodies were measured with the ring-test over six months after the first injection. The highest.titers of IgY was 1/2,560 at 2 months after the first injection. Approximately 60-80 mg of IgY were obtained from one egg yolk. Using the IgY, 1ng/ml of purified PMMoV was detected with the indirect ELISA. Gelrite gel double diffusion test, ELISA and tissue immuno-binding assay employing IgY gave similar sensitivity and specificity to those of IgG developed in rabbit. Therefore, the IgY which can be obtained in large quantities from a chicken, might be useful for the antibody production and the serology of plant viruses.

• Korean Journal of Poultry Science
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• v.31 no.1
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• pp.37-44
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• 2004

It has been recognized that the hen, like its mammalian counterparts, provides young chicks with antibodies as protection against hostile invaders. This system facilitates the transfer of specific antibodies from serum to egg yolk, and provides a supply of antibodies called immunoglobulin Y(IgY) to the developing embryo and the hatched chick. The protection against pathogens that the relatively immune-incompetent newly hatched chick has, is through transmission of antibodies from the mother via the egg. Egg yolk, therefore, can be loaded with a large amount of IgY against pathogens which can immobilize the existing or invading pathogens during the embryo development or in day-old chicks. Thus, the immunization of laying hens to various pathogens results in production of different antigen-specific IgY in eggs. Egg yolk contains 8∼20 mg of jmmunoglobulins (IgY) per ml or 136∼340 mg per yolk suggesting that more than 30 g of IgY can be obtained from one immunized hen in a year. By immunizing laying hens with antigens and collecting IgY from egg yolk, low cost antibodies at less than $10 per g compared to more than $20,000 per g of mammalian IgG can be obtained. This IgY technology opens new potential market applications in medicine, public health, veterinary medicine and food safety. A broader use of IgY technology could be applied as biological or diagnostic tool, nutraceutical or functional food development, oral-supplementation for prophylaxis, and as pathogen-specific antimicrobial agents for infectious disease control. This paper has emphasized that when IgY-loaded chicken eggs are produced and consumed, the specific antibody binds, immobilizes and consequently reduces or inhibits the growth or colony forming abilities of microbial pathogens. This concept could serve as an alternative agent to replace the use of antibiotics, since today, more and more antibiotics are less effective in the treatment of infections, due to the emergence of drug-resistant bacteria.

• Korean Journal of Veterinary Research
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• v.38 no.4
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• pp.829-836
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• 1998

Immunogenicity of Escherichia coli pilus and LT were evaluated in 20-week-old hens. The antigens were consisted of K88, K99, 987p pilus and heat labile toxin purified from enterotoxigenic Escherichia coli. The durations of antibody titers in sera and egg yolk were investigated by an enzyme-linked immunosorbent assay(ELISA). After first inoculation, antibody titers in sera reached at peak 2 weeks postinoculation. However, peak antibody titers in egg yolk were detected 4 weeks postinoculation, indicating that transfer of immunoglobulin from serum to egg yolk took about two weeks period. Although there were slight reduction in titers, the specific antibodies in egg yolk lasted up to 3 months. Immune responses against monovalent and combined antigens were showed as almost same patterns. The transfer rate of antibodies from serum to egg yolk didn't show any significant differences among three pilus antigens in this study. Considering the concentrations of antigens in each inoculated group, multivalent antigens containing heat labile toxin of E coli were found to be more immunogenic than monovalent antigen in producing specific antibodies. From this experiment, it was demonstrated that multivalent antigens containing three pilus and heat labile toxin could be a promising candidate for the production of egg yolk antibodies for prophylactic use in preventing swine colibacillosis in future.

• Journal of agriculture & life science
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• v.46 no.2
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• pp.107-114
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• 2012

The present study evaluated the potential use of immunoglobulin prepared from egg yolk of chickens immunized with Escherichia coli K88 (IgY-Ec) in the control of E. coli K88 infection in RAW 264.7 murine macrophage. The binding activity of IgY-Ec against E. coli K88 surface protein was more specific and increased than control IgY. In infection assay of E. coli in macrophage, the specific IgY-Ec to E. coli K88 remarkably inhibited the phagocytic activity comparing to nonspecific IgY (p<0.001). In adherence assay, bacterial adhesion on macrophage cells was definitely reduced by preincubation of IgY-Ec compared with nonspecific IgY (p<0.05). These findings suggested that IgY-Ec have the protective effects against pathogens and IgY-based diets may have potential benefits for preventing or treating various infections in domestic animals.

• Food Science of Animal Resources
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• v.33 no.5
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• pp.587-594
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• 2013

Egg consumption has been limited to avoid cardiovascular diseases such as atherosclerosis and hyperlipidemia, because the yolk contains high levels of cholesterol. This study was conducted to evaluate the effect of the water-soluble component of egg-yolk on the growth efficiency, immune modulation, and changes in serum lipid levels in BALB/c mice. A total 5 wk old 120 BALB/c male mice were divided into 4 groups and were fed 0, 2, 10, and 20 mg/d water-soluble egg yolk extract (WSEYE) for 5 wk. Water-soluble egg yolk extract (WSEYE) uptake resulted in a significant reduction in daily weight gain and feed efficiency rate (FER). The mouse groups treated with 2 and 20 mg/d WSEYE showed a significant increase in populations of monocytes at the third wk and B-lymphocyte activity at the fifth wk. In addition, WSEYE uptake did not influence serum immunoglobulin E levels. In serum lipid-profile studies, treatment of WSEYE did not alter total cholesterol and low-density lipoprotein levels; however, blood triglyceride levels were significantly diminished in mice treated with 2 mg/d at the third wk (p<0.05), and the level of high-density lipoprotein was significantly increased in the mice group treated with 2 and 10 mg/d WSEYE after 5 wk (p<0.05). Taken together, the data demonstrate the beneficial effects of WSEYE in the diet on immune modulation and serum lipid profiles in mouse models; therefore, this study suggests that ingestion of water-soluble fraction of egg yolk might not be related to the increased risk of heart disease, but can be an excellent candidate for maintaining health.

• Microbiology and Biotechnology Letters
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• v.25 no.6
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• pp.612-616
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• 1997

IgY (egg yolk immunoglobulin) against Helicobacter pylori was produced by immunizing hen with some Helicobacter pylori antigens. H. pylori whole cell, whole cell lysates, partially purified urease and p54 protein, which showed high antigenicity in mice, were used as immunogens. Four hens were immunized with these immunogens three times. IgY was purified from immunized egg yolk with polyethylene glycol (M.W. 8000) and its anti-H. pylori titer was determined by enzyme linked immunosorbent assay (ELISA). The anti-H. pylori titer reached peak at 8 weeks and was maintained over 20 weeks. H. pylori cells were agglutinated with these purified IgY and the specificity of these purified IgY was detected with immunoblotting.