Two groups of hens(control and immunization group)were arranged in an experimental design with an immunization schedule of 3 injections of BSA antigen.IgY antibodies were extracted from egg yolks by two precipitation ...Two groups of hens(control and immunization group)were arranged in an experimental design with an immunization schedule of 3 injections of BSA antigen.IgY antibodies were extracted from egg yolks by two precipitation processes(chloroform and polyethylene glycol precipitates)and quantified using a standard curve of protein concentration.The purification of IgY was confirmed by SDS-PAGE.Total protein extracted from egg yoks were less contaminated with yellow pigments(lutein and zeaxanthin)nd by using chloroform precipitate.The 2 week post-immunization,IgY-1 concentration increased respectively to 3903±726μg.ml(chloroform-1 extraction process)and 2937±294μg.ml(PEG extraction process)(P<rd 0.01).After 3 immunization,IgY level obtaining from in immunization group extracted by chloroform process(6633±1166μg.ml-1)increased 2.7 times higher than that in control group(2482±414μg.ml-1).Whereas IgY concentrations obtained from PEG extraction process were not significantly different between the experimental group and control group.Chloroform and PEG precipitation methods had the same protein profile on the SDS-PAGE.IgY antibody was identified by the presence of bands corresponding with IgY heavy chain(67-70 kDa)and IgY light chain(25 kDa)for both precipitation processes.展开更多
Specific immunoglobulin (IgY) from egg yolk against Aeromonas hydrophila was produced by immunization of White Leghorn hens with formalin-killed whole cells of A. hydrophila. ELISA test using A. hydrophila as the coat...Specific immunoglobulin (IgY) from egg yolk against Aeromonas hydrophila was produced by immunization of White Leghorn hens with formalin-killed whole cells of A. hydrophila. ELISA test using A. hydrophila as the coating antigen revealed that the specific antibody titer started to increase in the egg yolk at the 13th day post-immunization (P/N=2.18), reached the peak at the 56th day (P/N=13.82), and remained at high level until day 133 (P/N=7.03). The antibody was purified by saturated ammonium sulphate with a recovery rate of 63.5%. The specific IgY inhibited the growth of A. hydrophila at a concentration of 1.0 mg/ml during the 18 h incubation. Pre-treatment of polyploid gibel carps Carassius auratus Gibelio with specific IgY had a protection rate of 60% (6/10) against challenge with A. hydrophila, while none of the fishes in the control groups receiving sterile phosphate buffered saline (PBS) or non-specific IgY survived the challenge. Treatment of fishes with the specific IgY 4 h after the challenge also had lower mortality (70%, 7/10), a 30% reduction against the control PBS or non-specific IgY groups (10/10). These results indicate that specific IgY antibodies could be obtained easily from hens immunized with an inactivated A. hydrophila and could provide a novel alternative approach to control of diseases in fishes caused by this organism.展开更多
The term“IgY technology”was introduced in the literature in the mid 1990s to describe a procedure involving immunization of avian species,mainly laying hens and consequent isolation of the polyclonal IgYs from the“...The term“IgY technology”was introduced in the literature in the mid 1990s to describe a procedure involving immunization of avian species,mainly laying hens and consequent isolation of the polyclonal IgYs from the“immune”egg yolk(thus avoiding bleeding and animal stress).IgYs have been applied to various fields of medicine and biotechnology.The present article will deal with specific aspects of IgY technology,focusing on the currently reported methods for developing,isolating,evaluating and storing polyclonal IgYs.Other topics such as current information on isolation protocols or evaluation of IgYs from different avian species are also discussed.Specific advantages of IgY technology(e.g.,novel antibody specificities that may emerge via the avian immune system)will also be discussed.Recent in vitro applications of polyclonal egg yolk-derived IgYs to the field of disease diagnosis in human and veterinary medicine through in vitro immunodetection of target biomolecules will be presented.Moreover,ethical aspects associated with animal well-being as well as new promising approaches that are relevant to the original IgY technology(e.g.,development of monoclonal IgYs and IgY-like antibodies through the phage display technique or in transgenic chickens)and future prospects in the area will also be mentioned.展开更多
Aim To study whether antivenom from laying hens can be used for the detection of venom antigens, Methods Chickens （white Leghorn） were immunized with detoxicated king cobra venom by formaldehyde and egg yolk immunog...Aim To study whether antivenom from laying hens can be used for the detection of venom antigens, Methods Chickens （white Leghorn） were immunized with detoxicated king cobra venom by formaldehyde and egg yolk immunoglobulin （IgY） isolated from yolk; IgY was labelled with the horseradish peroxidase （HRP）. Experimental condition and parameters were determined by chessboard test. The specificity, sensitivity, precision, and stability of this method were assayed in the experiment. Results This method could detect as low as 32 μg· L^-1 of the king cobra antigens. A good linear relation was found within 32 ～ 750 μg· L^-1 of king cobra venom concentrations （ r = 0. 963）. There was no cross reactivity for the reagents with Agkistrodon acutus Guenther venom or Vipera russelli siamensis Smith venom;slight cross reactivity .with Bungarus multicinctus Blyth venom or Bungarus fasciatus Chmeider venom; and notable cross reactivity with cobra venom. The average intra-assay relative standard deviation （RSD） was 1% - 3%, and the inter-assay RSD was less than 8%. The reagents （including IgY and HRP-IgY） were stable; no differences （P 〉 0.05） were observed for the detection of venom antigens when the reagents were stored at 37 ℃ up to 6 d. Conclusion IgY is a good reagent for diagnosis of snakebite after eliminating the genus cross reactivity.展开更多
Layer chickens were immunized with three species of inactivated orthopox virus （vaccinia virus, calpox virus and cowpox virus）. Antibodies （IgY） were purified from egg yolks by improved polyethylene glycol precipi...Layer chickens were immunized with three species of inactivated orthopox virus （vaccinia virus, calpox virus and cowpox virus）. Antibodies （IgY） were purified from egg yolks by improved polyethylene glycol precipitation. The development of IgY directed against orthopox virus antigens was followed by immunofluorescence assay, plaque reduction neutraliztion test and immunoelectron microscopy. Cross-reactivity of two IgY antibodies with cells infected by the different strains of the pox viruses was also investigated by different methods （immunofluorescence assay, plaque reduction neutraliztion test and Western blot）. Even in very high dilutions in immunofluorescence assay （titres up to 1：10^6 and 1：10^5, respectively） and persisted on a plateau over 10 months after four booster injections, it was showed that anti-vaccinia virus IgY and anti-calpox virus IgY were positive. Neutralizing activity and ultra-structural detection of antigen with gold-labelled antibodies were respectively observed in plaque reduction neutralization test and immunoelectron microscopy. Western blot analysis revealed specific binding of IgY to virus proteins. Thus, there was cross-reactivity between different orthopox viruses. Finally, orthopox virns-specific IgY antibodies bounded magnetic beads （Dynabead） were used to concentration of orthopox viruses. This study suggests that anti-pox virus IgY could serve as a useful tool for orthopox viruses diagnosis.展开更多
In recent years, the use of in-feed antibiotics for growth and disease prevention in livestock production has been under severe scrutiny. The use and misuse of in-feed antibiotics has led to problems with drug residue...In recent years, the use of in-feed antibiotics for growth and disease prevention in livestock production has been under severe scrutiny. The use and misuse of in-feed antibiotics has led to problems with drug residues in animal products and increased bacterial resistance. Chicken egg yolk antibodies （IgY） have attracted considerable attention as an alternative to antibiotics to maintain swine health and performance. Oral administration of IgY possesses many advantages over mammalian IgG such as cost-effectiveness, convenience and high yield. This review presents an overview of the potential to use IgY immunotherapy for the prevention and treatment of swine diarrhea diseases and speculates on the future of IgY technology. Included are a review of the potential applications of IgY in the control of enteric infections of either bacterial or viral origin such as enterotoxigenic Escherichia coil, Salmonella spp., rotavirus, porcine transmissible gastroenteritis virus, and porcine epidemic diarrhea virus. Some potential obstacles to the adoption of IgY technology are also discussed.展开更多
Objective:To identify unique immunogenic epitopes of Zika virus non-structural 1(NS1)antigen and produce immunoglobulin Y(IgY)for potential use in he diagnosis of of Zika virus infection.Methods:Immunogenic epitopes w...Objective:To identify unique immunogenic epitopes of Zika virus non-structural 1(NS1)antigen and produce immunoglobulin Y(IgY)for potential use in he diagnosis of of Zika virus infection.Methods:Immunogenic epitopes were identified using in silico B-cell epitope prediction.A synthetic peptide analog of the predicted epitope was used to induce antipeptide IgY production in hens which was purified using affinity chromatography.Presence of purified IgY and its binding specificity were performed by gel electrophoresis and ELISA,respectively.Results:Out of the nine continuous epitopes identified,the sequence at position 193-208(LKVREDYSLECDPAVI)was selected and used to produce anti-peptide IgY.The produced IgY was found to bind to the synthetic analog of the Zika virus NS1 immunogenic epitope but not to other flaviviruses and random peptides from other pathogens.Conclusions:In this study,we identified an immunogenic epitope unique to Zika virus that can be used to develop a serodiagnostic tool that specifically detect Zika virus infection.展开更多
卵黄抗体(Immunoglobulin of yolk,IgY)因在水生动物疾病中具有被动免疫疗效而受到广泛的关注。本文综述了IgY基本结构和理化性质的研究进展,重点介绍了IgY在水生动物细菌和病毒感染防控、治疗中的潜在应用,为今后IgY的研究和水产养殖...卵黄抗体(Immunoglobulin of yolk,IgY)因在水生动物疾病中具有被动免疫疗效而受到广泛的关注。本文综述了IgY基本结构和理化性质的研究进展,重点介绍了IgY在水生动物细菌和病毒感染防控、治疗中的潜在应用,为今后IgY的研究和水产养殖中的应用提供参考。展开更多
基金the scientific research fund of Nong Lam University,Ho Chi Minh City for giving the grant to this study.
文摘Two groups of hens(control and immunization group)were arranged in an experimental design with an immunization schedule of 3 injections of BSA antigen.IgY antibodies were extracted from egg yolks by two precipitation processes(chloroform and polyethylene glycol precipitates)and quantified using a standard curve of protein concentration.The purification of IgY was confirmed by SDS-PAGE.Total protein extracted from egg yoks were less contaminated with yellow pigments(lutein and zeaxanthin)nd by using chloroform precipitate.The 2 week post-immunization,IgY-1 concentration increased respectively to 3903±726μg.ml(chloroform-1 extraction process)and 2937±294μg.ml(PEG extraction process)(P<rd 0.01).After 3 immunization,IgY level obtaining from in immunization group extracted by chloroform process(6633±1166μg.ml-1)increased 2.7 times higher than that in control group(2482±414μg.ml-1).Whereas IgY concentrations obtained from PEG extraction process were not significantly different between the experimental group and control group.Chloroform and PEG precipitation methods had the same protein profile on the SDS-PAGE.IgY antibody was identified by the presence of bands corresponding with IgY heavy chain(67-70 kDa)and IgY light chain(25 kDa)for both precipitation processes.
基金Project (No. 2004C26026) supported by the Science and Technology Department of Zhejiang Province, China
文摘Specific immunoglobulin (IgY) from egg yolk against Aeromonas hydrophila was produced by immunization of White Leghorn hens with formalin-killed whole cells of A. hydrophila. ELISA test using A. hydrophila as the coating antigen revealed that the specific antibody titer started to increase in the egg yolk at the 13th day post-immunization (P/N=2.18), reached the peak at the 56th day (P/N=13.82), and remained at high level until day 133 (P/N=7.03). The antibody was purified by saturated ammonium sulphate with a recovery rate of 63.5%. The specific IgY inhibited the growth of A. hydrophila at a concentration of 1.0 mg/ml during the 18 h incubation. Pre-treatment of polyploid gibel carps Carassius auratus Gibelio with specific IgY had a protection rate of 60% (6/10) against challenge with A. hydrophila, while none of the fishes in the control groups receiving sterile phosphate buffered saline (PBS) or non-specific IgY survived the challenge. Treatment of fishes with the specific IgY 4 h after the challenge also had lower mortality (70%, 7/10), a 30% reduction against the control PBS or non-specific IgY groups (10/10). These results indicate that specific IgY antibodies could be obtained easily from hens immunized with an inactivated A. hydrophila and could provide a novel alternative approach to control of diseases in fishes caused by this organism.
文摘The term“IgY technology”was introduced in the literature in the mid 1990s to describe a procedure involving immunization of avian species,mainly laying hens and consequent isolation of the polyclonal IgYs from the“immune”egg yolk(thus avoiding bleeding and animal stress).IgYs have been applied to various fields of medicine and biotechnology.The present article will deal with specific aspects of IgY technology,focusing on the currently reported methods for developing,isolating,evaluating and storing polyclonal IgYs.Other topics such as current information on isolation protocols or evaluation of IgYs from different avian species are also discussed.Specific advantages of IgY technology(e.g.,novel antibody specificities that may emerge via the avian immune system)will also be discussed.Recent in vitro applications of polyclonal egg yolk-derived IgYs to the field of disease diagnosis in human and veterinary medicine through in vitro immunodetection of target biomolecules will be presented.Moreover,ethical aspects associated with animal well-being as well as new promising approaches that are relevant to the original IgY technology(e.g.,development of monoclonal IgYs and IgY-like antibodies through the phage display technique or in transgenic chickens)and future prospects in the area will also be mentioned.
文摘Aim To study whether antivenom from laying hens can be used for the detection of venom antigens, Methods Chickens （white Leghorn） were immunized with detoxicated king cobra venom by formaldehyde and egg yolk immunoglobulin （IgY） isolated from yolk; IgY was labelled with the horseradish peroxidase （HRP）. Experimental condition and parameters were determined by chessboard test. The specificity, sensitivity, precision, and stability of this method were assayed in the experiment. Results This method could detect as low as 32 μg· L^-1 of the king cobra antigens. A good linear relation was found within 32 ～ 750 μg· L^-1 of king cobra venom concentrations （ r = 0. 963）. There was no cross reactivity for the reagents with Agkistrodon acutus Guenther venom or Vipera russelli siamensis Smith venom;slight cross reactivity .with Bungarus multicinctus Blyth venom or Bungarus fasciatus Chmeider venom; and notable cross reactivity with cobra venom. The average intra-assay relative standard deviation （RSD） was 1% - 3%, and the inter-assay RSD was less than 8%. The reagents （including IgY and HRP-IgY） were stable; no differences （P 〉 0.05） were observed for the detection of venom antigens when the reagents were stored at 37 ℃ up to 6 d. Conclusion IgY is a good reagent for diagnosis of snakebite after eliminating the genus cross reactivity.
文摘Layer chickens were immunized with three species of inactivated orthopox virus （vaccinia virus, calpox virus and cowpox virus）. Antibodies （IgY） were purified from egg yolks by improved polyethylene glycol precipitation. The development of IgY directed against orthopox virus antigens was followed by immunofluorescence assay, plaque reduction neutraliztion test and immunoelectron microscopy. Cross-reactivity of two IgY antibodies with cells infected by the different strains of the pox viruses was also investigated by different methods （immunofluorescence assay, plaque reduction neutraliztion test and Western blot）. Even in very high dilutions in immunofluorescence assay （titres up to 1：10^6 and 1：10^5, respectively） and persisted on a plateau over 10 months after four booster injections, it was showed that anti-vaccinia virus IgY and anti-calpox virus IgY were positive. Neutralizing activity and ultra-structural detection of antigen with gold-labelled antibodies were respectively observed in plaque reduction neutralization test and immunoelectron microscopy. Western blot analysis revealed specific binding of IgY to virus proteins. Thus, there was cross-reactivity between different orthopox viruses. Finally, orthopox virns-specific IgY antibodies bounded magnetic beads （Dynabead） were used to concentration of orthopox viruses. This study suggests that anti-pox virus IgY could serve as a useful tool for orthopox viruses diagnosis.
基金supported by funds from the National Natural Science Foundation of China(310010533037105330871806)
文摘In recent years, the use of in-feed antibiotics for growth and disease prevention in livestock production has been under severe scrutiny. The use and misuse of in-feed antibiotics has led to problems with drug residues in animal products and increased bacterial resistance. Chicken egg yolk antibodies （IgY） have attracted considerable attention as an alternative to antibiotics to maintain swine health and performance. Oral administration of IgY possesses many advantages over mammalian IgG such as cost-effectiveness, convenience and high yield. This review presents an overview of the potential to use IgY immunotherapy for the prevention and treatment of swine diarrhea diseases and speculates on the future of IgY technology. Included are a review of the potential applications of IgY in the control of enteric infections of either bacterial or viral origin such as enterotoxigenic Escherichia coil, Salmonella spp., rotavirus, porcine transmissible gastroenteritis virus, and porcine epidemic diarrhea virus. Some potential obstacles to the adoption of IgY technology are also discussed.
文摘Objective:To identify unique immunogenic epitopes of Zika virus non-structural 1(NS1)antigen and produce immunoglobulin Y(IgY)for potential use in he diagnosis of of Zika virus infection.Methods:Immunogenic epitopes were identified using in silico B-cell epitope prediction.A synthetic peptide analog of the predicted epitope was used to induce antipeptide IgY production in hens which was purified using affinity chromatography.Presence of purified IgY and its binding specificity were performed by gel electrophoresis and ELISA,respectively.Results:Out of the nine continuous epitopes identified,the sequence at position 193-208(LKVREDYSLECDPAVI)was selected and used to produce anti-peptide IgY.The produced IgY was found to bind to the synthetic analog of the Zika virus NS1 immunogenic epitope but not to other flaviviruses and random peptides from other pathogens.Conclusions:In this study,we identified an immunogenic epitope unique to Zika virus that can be used to develop a serodiagnostic tool that specifically detect Zika virus infection.
