• Korean Journal of Poultry Science
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• v.41 no.1
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• pp.15-20
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• 2014

Fowl adenovirus (FAV) is an important cause of several diseases, which result in considerable economic losses to the poultry farm. An outer capsid protein of FAV, fiber 2 is essential for virus growth, assembly or spread. This study was performed to produce about 22 kDa of recombinant fiber 2 protein and to immunize in laying hens to acquire the specific IgY antibody against the recombinant fiber 2. Laying hens were immunized with the recombinant fiber 2 intramuscularly in the breast muscle by injection 4 times at intervals of three weeks. At 12 weeks, serum- and egg yolk-antibody titers of hens against fiber 2 were increased up to 430,000 and 414,000, respectively. The recombinant fiber 2 could be recognized be the anti-His monoclonal antibody. Anti-fiber 2-IgY antibody could recognize the fiber 2 specifically in western blot analysis. These results suggested that the recombinant fiber 2 antigen could be used as an immunogen to elicit IgY antibody against fiber 2 and the anti-fiber 2-IgY could neutralize fowl adenovirus fiber 2 effectively.

• Asian-Australasian Journal of Animal Sciences
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• v.19 no.11
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• pp.1665-1670
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• 2006

The objective of this research was to provide the characterization and method for producing anti-E. coli O157:H7 antibodies in egg-laying hens and to determine if the antibody can restrain the proliferation of E. coli O157:H7 in-vitro. Selected antigenic fractions (whole cell, outer membrane protein and lipopolysaccharide (LPS)) from E. coli O157:H7 were injected to hens in order to produce anti-E. coli O157:H7 antibodies. The immune response and the egg yolk antibodies of laying hens against the whole cell, outer membrane protein and LPS antigens were monitored by ELISA. The level of antibodies against whole cell antigen monitored through ELISA sharply increased after the initial immunization, and it was found to be maximum on day 49 however, the level was maintained up to day 70. Antibodies (5 mg/ml) directed against the whole cell inhibited E. coli proliferation 10-13 times more than outer membrane protein or LPS. The antibody response against the whole cell antigens appeared to have higher activity in restraining the proliferation of E. coli O157:H7 than antibody against outer membrane protein or LPS. Results reflected that increasing the IgY's in the egg yolk could prevent greater economic losses due to human and animal health from pathogenic bacteria i.e. E. coli O157:H7.

• Korean Journal of Food Science and Technology
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• v.30 no.5
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• pp.1029-1034
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• 1998

Chick antibodies (IgY) raised against Streptococcus mutans (serotype c) were separated from egg yolk and their properties were investigated. The purity of IgY extracts prepared by the method of ${\lambda}-carrageenan$, $gammaYolk^{TM}$, and $EGGstract^{TM}$ was 20%, 46%, and 48%, respectively, and the yields of IgY extracts from a gram yolk were 11. 3 mg, 1.7 mg, and 1.8mg, respectively. Quantitative immunoprecipitation test showed that specific IgY content of crude IgY prepared by ${\lambda}-carrageenan$ method was 12.2%, which means that 0.85 g of crude IgY from an egg yolk (15 g) contains about 100 mg of specific IgY. When the reactivity of the specific IgY towards 3 caries-inducing strains (serotype: b, c, f) was examined, the strains cultured in sucrose-added medium showed higher reactivity (the orders were c(+), f(+), b(+)) than those cultured in sucrose-free medium. Heat and pH stability of specific IgY was good, for crude IgY contained 50% of antibody activity after heat treatment at $70^{\circ}C$ for 5 min and they were stable at pH $4{\sim}8$.

• Food Science of Animal Resources
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• v.24 no.2
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• pp.182-188
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• 2004

To determine the concentration of Lactoperoxidase (LPO), an indirect enzyme-linked immunosorbant assay(ELISA) was developed. Anti-LPO egg yolk immunoglobulin(IgY) was transferred to egg yolk by immunizing of Brown hens with LPO. The titer of purified anti-LPO IgY was 1: 520,000. The immunological response of anti- LPO IgY with ${\alpha}$-lactalbumin, ${\beta}$-lactoglobulin, casein and lysozyme were evaluated, resulting that the anti-LPO IgY found to be a specific antibody toward LPO and no cross-reaction was observed against ${\alpha}$-lactalbumin, ${\beta}$-lactoglobulin, casein, and lysozyme in double immunodiffusion test and ELISA test. In indirect ELISA method, coating concentration of LPO and dilution rate of anti-LPO IgY was 0.25$\mu\textrm{g}$/mL and 1:8,000 respectively. Sensitivity in the standard curve of LPO was ranged from 0.01 to 1$\mu\textrm{g}$/mL using anti-LPO IgY.

• Proceedings of the Korea Society of Poultry Science Conference
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• 2003.07b
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• pp.37-54
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• 2003

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 immuno-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 immunoglobulins (IgY) per $m\ell$ 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. nut-raceutical 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.

• Food Science of Animal Resources
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• v.32 no.6
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• pp.706-712
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• 2012

The present study was aimed to produce a chicken polyclonal antibody against Cronobacter muytjensii and to develop an immunoassay for its detection. Purification of anti-C. muytjensii IgY from egg yolk was accomplished using various methods such as water dilution and salt precipitation. As a result, sodium dodecyl sulfate-polyacrylamide gel electrophoresis produced two bands around 30 and 66 kDa, corresponding to a light and a heavy chain, respectively. Indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) was performed to determine the effectiveness of the chicken IgY against C. muytjensii. The optimum conditions for detecting C. muytjensii by indirect ELISA and checkerboard titration of the antigen revealed an optimum average absorbance at the concentration of 18 ${\mu}g/mL$, having ca. $10^8$ coated cells per well. The anti-C. muytjensii IgY antibody had high specificity for C. muytjensii and low cross-reactivity with other tested pathogens. In this assay, no cross-reactivity was observed with the other genera of pathogenic bacteria including Escherichia coli O157:H7, Salmonella Typhimurium, Staphylococcus aureus, Bacillus cereus, Enterobacter aerogenes, Salmonella Enteritidis and Listeria monocytogenes. In addition, detection of C. muytjensii in infant formula powder showed a low matrix effect on the detection curve of IC-ELISA for C. muytjensii, with similar detection limit of $10^5$ CFU/mL as shown in standard curve. These findings demonstrate that the developed method is able to detect C. muytjensii in infant formula powder. Due to the stable antibody supply without sacrificing animals, this IgY can have wide applications for the rapid and accurate detection of C. muytjensii in dairy foods samples.

• Korean Chemical Engineering Research
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• v.50 no.5
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• pp.866-873
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• 2012

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 and four-zone SMB chromatography. Before performing SMB experiments, batch chromatography simulation parameters and adsorption isotherms were obtained. The parameters of batch chromatography were used to simulate SMB using Aspen chromatography. To compare three-zone and four-zone SMB chromatography, simulations in $m_2-m_3$ plane on the triangle theory were carried out. In terms of concentration and purity of both IgY and other lipoproteins, 3-zone SMB process is considered as ideal at the vertex of triangle ($m_2$, $m_3$=0.1, 1.1). 4-zone SMB yields the highest IgY purity at the coordinate ($m_2$, $m_3$=0.06, 0.5), which is the pure raffinate region. In 3-zone SMB without recycle, other lipoproteins in extract are largely affected in purity by small shift from the vertex of triangle ($m_2$, $m_3$=0.1, 1.1).

• Journal of Microbiology and Biotechnology
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• v.23 no.4
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• pp.527-533
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• 2013

Polyclonal antibodies labeled with a tracer have been commonly used as secondary antibodies in immunochemical assays to quantify the concentration of antibody-antigen complexes. The majority of these antibodies conjugated with a tracer are commercially available, with the exception of few untouched targets. This study focused on the production and application of mouse anti-quail IgY as an intermediate antibody to link between quail egg yolk IgY and goat anti-mouse IgG-HRP as primary and secondary antibodies, respectively. Subsequently, the produced mouse anti-quail IgY was labeled with horseradish peroxidase (HRP) and its efficiency on enzyme linked immunosorbent assay (ELISA) was compared with that of commercial rabbit anti-chicken IgY-HRP. As an intermediate antibody, mouse anti-quail IgY was successfully produced with good affinity and sensitivity (1:10,000) to the primary and secondary antibodies. Subsequently, mouse anti-quail IgY was effectively conjugated with HRP enzyme, resulting in a secondary antibody with good sensitivity (1:10,000) to quail anti-V. parahaemolyticus and V. vulnificus IgY. The detection limit was $10^5$ CFU/ml for both V. parahaemolyticus and V. vulnificus. The efficiency of the produced conjugate to detect quail IgY on ELISA was comparable to that of the commercial rabbit anti-chicken IgY-HRP, and hence the produced and labeled mouse anti-quail IgY-HRP can be used as a secondary antibody to detect any antibody produced in quail.

• Microbiology and Biotechnology Letters
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• v.50 no.3
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• pp.430-435
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• 2022

The infectious bursal disease (IBD) is a highly contagious and acute poultry disease caused by Birnavirus. However, the vaccination is the only disease prevention, but several factors impeded vaccine development. Thus, a need for time to develop a novel technique for managing and treating respiratory diseases in poultry birds. Passive immunization is a hope and a possible alternative used in birds to meet this need. The current research attempted to produce egg yolk-based polyclonal antibodies against the IBD virus. The benefits of IgY include ease of extraction, lack of reaction with mammalian Fc receptors, and low production cost. Commercial layers were immunized with inactivated IBD virus subcutaneously according to the treatment regimen. The eggs were gathered daily, and yolk antibodies were extracted with the ammonium sulfate precipitation technique. The use of an indirect hemagglutination test demonstrated that IgY was IBD-specific. Until the end of the experiment, the specific IgY immunoglobulins did not lose activity when stored at 4℃. The specific immunoglobulin (IgY) treated challenged birds were demonstrated 92% recovery in comparison to the control group. The study implies that the IBDV specific IgY is an easily prepared and rich source of antibodies and offers an alternative therapeutic agent to cure IBD-infected birds.

• Korean Journal of Veterinary Research
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• v.36 no.4
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• pp.895-902
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• 1996

산란계에 불활화 개 파보바이러스 백신을 근육내로 1주 간격으로 4회 접종하여 면역화시키고 최종 접종 2주후에 채란하여 $4^{\circ}C$에 보관하며 사용하였다. 난황항체는 5% HPMCP를 이용하여 분리하였고 0.5% HPMCP 용액은 lipid 침전에 매우 효과적이었으며 희석배수 10배에서 투명한 상층액을 나타내었다. 1차분리한 상층액의 단백질 농도는 2.5mg/ml이었고 최종 단백질 용액의 경우는 26.53mg/ml이었다. SDS-PAGE 전기영동상에서 분자량 60~70 KD 및 30~40 KD의 2 band가 나타났으며 non-reducing 전기영동에서는 닭 혈청 IgG와 같은 120~160 KD의 분자량을 보인 band가 각각의 분리용액에서 나타났다. 난황 항체의 개 파보바이러스에 대한 혈구응집억제반응 항체역가는 혈청의 역가에 비해 1주의 차이를 주며 증가했으며 난황 항체는 1:640에서 1:2560, 혈청은 1:640에서 1:5120을 나타내었다.