• Journal of Animal Science and Technology
• /
• v.46 no.3
• /
• pp.495-502
• /
• 2004

Intimin, the product of eae gene in EHEC O157:H7, is required for intimate adherence. In this study, the C-terminaI region(281 amino acids) of the EHEC OI57:H7 intimin were expressed as a protein fusion with (His)$_6$ which was used to raise antiserum in rabbits. The antiserum reacted in western blot with a 94kDa outer membrane protein of EHEC O157:H7. It was observed that the antibody titers both in egg yolk and serum appeared in 2${\sim}$4 weeks after immunization with fusion protein. At the time of 8 weeks, the titre of egg yolk was found to be higher than that of sera. According to the results of neutralization test, chicken egg-yolk antibody(lgY) against the recombinant intimin strongly reacted to EHEC O157:H7. We conclude that a truncated recombinant intimin could be used as an immunogen to elicit antibody(lgY) against O157:H7.

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

• 한국생물공학회:학술대회논문집
• /
• 2005.10a
• /
• pp.571-571
• /
• 2005

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

• Korean Journal of Food Science and Technology
• /
• v.30 no.5
• /
• pp.1029-1034
• /
• 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
• /
• v.24 no.2
• /
• pp.182-188
• /
• 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.

• Journal of Microbiology and Biotechnology
• /
• v.29 no.2
• /
• pp.311-320
• /
• 2019

Fusobacterium nucleatum is a morbific agent in periodontitis and halitosis. Egg yolk antibody (IgY) was obtained from egg yolks from chickens stimulated with F. nucleatum. This study was to assess the effectiveness of IgY on periodontitis and halitosis caused by F. nucleatum in vitro and in vivo. The growth of F. nucleatum was inhibited (p <0.05) by different concentrations of IgY in vitro and the results of a Halimeter show volatile sulfur compounds (VSCs) were reduced to $904{\pm}57ppb$ at a concentration 40 mg/ml of IgY. The changes of fatty acids of F. nucleatum were determined using GC-MS. The scores for odor index of rat saliva were decreased. The major constituent of volatile organic compounds (VOCs) including short-chain acids decreased 46.2% in 10 mg/ml IgY, ammonia decreased 70% in 40 mg/ml IgY, while aldehydes and olefine ketones were almost unchanged. The ELISA assay revealed that IL-6 and TNF-${\alpha}$ were decreased after 4 weeks' IgY treatment. Morphometric (X-ray) and histological analyses (HE) showed that IgY reduced alveolar bone loss and collagen fibers became orderly in rat models. As a result, IgY may have the potential to treat periodontitis and halitosis.

• Animal cells and systems
• /
• v.5 no.1
• /
• pp.85-90
• /
• 2001

Yolk platelets, one of the main food stores in the embryonic development, are composed of proteins. However, little is known about the identity of proteins utilized at certain stages of embryogenesis. In this study, we followed the fates of embryonic storage proteins by using an anti-polyubiquitin monoclonal antibody (mAB) as a probe. The mAb recognized the major storage proteins of Drosophila, Xenopus and chicken eggs. In the Drosophila embryo, the mAb-reactive 45-kDa protein was not used until stage 11 but was used up at stage 16 when the embryo completed segmentation. In the Xenopus embryo, the mAb-reactive 111 kDa protein was mostly utilized between stages 42 and 45 implying that the protein might be an energy source used just prior to feeding on food. By N-terminal sequencing the storage protein of Xenopus embryo was identified as a lipovitellin 1. This study confirms that storage proteins are used almost simultaneously at certain stages of embryogenesis and that vitellogenin 1 is the last storage protein in Xenopus embryogenesis.

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

• Asian-Australasian Journal of Animal Sciences
• /
• v.17 no.8
• /
• pp.1139-1144
• /
• 2004

In Exp. 1, a total of 36 pigs (6.55$\pm$0.10 kg average initial body weight and 21 d average age) were used in a 14 d growth study to determine the effects of replacing spray-dried plasma protein (SDPP) with spray-dried egg protein containing specific egg yolk antibody (SDEP) on growth performance and nutrient digestibility in weaned pigs. The pigs were blocked by weight and assigned to treatments based on sex. There were three pigs per pen and four pens per treatment. Dietary treatments were 0, 3, or 6% SDEP and contained 6, 3, or 0% SDPP, respectively. Through the entire experimental period, average daily gain (ADG), average daily feed intake (ADFI), and gain/feed tended to decrease as the concentration of SDEP increased in the diets. However, there were no significant differences among the treatments (p>0.05). As the addition of SDEP in the diets increased, apparent digestibilities of dry matter (DM) and nitrogen (N) were decreased without significant (p>0.05). For Exp. 2, 36 pigs (2.63$\pm$0.04 kg average initial body weight and 10 d average age) were used in a 14 d growth study to determine the effects of antibiotic replacement with SDEP on growth performance and nutrient digestibility in early-weaned pigs. The pigs were blocked by weight and assigned to treatments based on sex. There were three pigs per pen and four pens per treatment. Dietary treatments included 1) ANTIBIOTIC (corn-dried whey-soybean meal based diet+0.08% antibiotics, 4 mg of tiamuline hydrogen fumarate; 10 mg of sulfadimidine per kg of complete diet), 2) SDEP0.1 (corndried whey-SBM based diet+0.1% SDEP), and 3) SDEP0.2 (corn-dried whey-SBM based diet+0.2% SDEP). ADG and gain/feed of pigs fed the SDEP0.2 diet were higher than for pigs fed the ANTIBIOTIC diet without significant (p>0.05). Pigs fed the diet with SDEP0.2 tended to have increased apparent digestibilities of DM and N compared to pigs fed the ANTIBIOTIC diet without significant (p>0.05). In conclusion, the dietary SDEP seemed to be partial replacing the SDPP portion of high nutrient dense diet for weaned pigs. Also, dietary SDEP seemed to be approximately 0.2% or more when the pigs fed the antibiotic-free diet for early-weaned pigs.