Extraction, Purification and invitro Challenging of IgY of Commercial Chicken Eggs with the Newcastle Disease (original) (raw)
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Atla-alternatives To Laboratory Animals, 2005
The production of antibodies (Abs) in chickens and the extraction of specific Abs from egg yolk (IgY Abs) are increasingly attracting the interest of the scientific community, as demonstrated by the significant growth of the IgY literature. This review offers detailed and comprehensive information about IgY-technology, including: a) possibilities for hen keeping in accordance with the Three Rs principles; b) new insights into the IgY transfer mechanism from blood to yolk as a biological basis for the technology; c) the comparative characteristics of IgY Abs and IgG Abs; d) the high efficacy of the technique, in view of the extraordinary amount of IgY Ab produced by one hen in one year (between 20g and 40g IgY in total); e) comparisons between the efficacies of IgY Abs and IgG Abs (rabbit, sheep, mouse) in several immunological assays; f) immunisation protocols, as well as the most commonly used IgY-extraction procedures; g) new possibilities for application in human and veterinary medicine, including strategies for the treatment of Helicobacter pylori infection or fatal intestinal diseases in children, particularly in poor countries, for reducing the use of antibiotics, and, in Asia and South America, for producing Abs against snake, spider and scorpion venoms; and h) the use of IgY Abs in various fields of research, also taking into consideration recent developments in South America (particularly Argentina and Cuba) and in Asia.
Production and purification of IgY antibodies from chicken egg yolk
Journal of Genetic Engineering and Biotechnology, 2018
The isolation of polyclonal antibodies from the serum of immunized mammals has significantly contributed to scientific research and diagnosis. The fact that recent technologies allow the production of antibodies in the yolk of eggs laid by hens, has led to the development of an alternative method for antibody generation that is less stressful to animals. As hens are kept under almost all their natural conditions, antibodies are isolated from the collected eggs; this technology is expected to become an interesting alternative to the conventionally serum-based techniques that eventually require to sacrifice the animal. Here we present a modified protocol for the isolation of IgY antibodies from immunized chickens and provide comparison between two chicken breeds in relative to IgY yield per egg. Our results show the possibility of generating large quantities of highly pure IgY from chicken eggs and also show large differences in the yield of IgY production between the two studied breeds. The results of this work indicate that IgY technology can be used for the production of primary antibodies for immunological work and disease diagnosis.
Production, characterization and application of monoclonal antibodies against chicken IgY
Veterinarski Arhiv, 2004
Monoclonal antibodies (Mabs) were produced against chicken IgY purified from egg yolks. Purity of the IgY was established using immunoelectrophoresis and reducing and non-reducing polyacrylamide gel electrophoresis. Five stable hybrid clones were characterized and Mabs, all belonging to IgG 1 isotype, were found to bind to the IgY heavy chain in Western blots. These clones did not cross-react with IgM in enzyme-linked immunosorbent assay (ELISA). One of the Mabs was used as a capture antibody in ELISA for estimation of IgY purified from egg yolk by different methods. Caprylic acid precipitation method gave the highest recovery of IgY and with high purity.
ILAR Journal, 2005
Polyclonal antibody production in mammals is generally associated with multiple injections of antigens and adjuvants and repeated blood sampling procedures. During the past 20 yr, the use of chickens instead of mammals for this purpose has increased. A major advantage of using birds is that the antibodies can be harvested from the egg yolk instead of serum, thus making blood sampling obsolete. In addition, the antibody productivity of an egg-laying hen is much greater than that of a similar sized mammal. This article focuses on the developments in oral immunization strategies for chickens that combined with the antibodies from the egg yolk, have great potential for active implementation of the three Rs (replacing, reducing, and refining the use of laboratory animals to the extent possible) in polyclonal antibody production schemes.
Antibodies have become essential tools in recent decades, with a wide range of applications in the laboratory and in human and veterinary medicine. The use of laying hens, instead of mammals, to obtain the necessary antibodies from the eggs is a major advance in terms of animal welfare because it makes blood sampling obsolete. However, the advantages of this technology are numerous, in addition to the animal welfare aspect. With a carefully designed immunization protocol, it is possible to enhance both the hen's immune response and its welfare during the process. This review puts forward recommendations how to do this and discusses recent approaches on improving the technology.
2009
Enteric infections caused by Salmonella remain a major public health burden worldwide. Poultry, particularly chickens, are known to be the main reservoir for this zoonotic pathogen. Therefore, the prevention and monitoring of Salmonella infection during the live phase may greatly reduce the contamination of poultry meat during slaughter and processing. With the ban on sub-therapeutic antibiotic usage in Europe and the increasingly strictness of the European legislation on food hygiene, passive immunization by oral administration of pathogen-specific hen egg yolk antibody (IgY) may be a useful and attractive alternative. This review offers summarized information about IgY production and the use of these antibodies for passive immunization, particularly in poultry. Keywords. Hen egg yolk antibodies, IgY production, passive immunization, enteric infections, feed additive. Les anticorps du jaune d'oeuf de poule (IgY), production et utilisation en immunisation passive contre les infections entériques bactériennes : une revue. Les infections entériques causées par Salmonella constituent un problème majeur de santé publique à travers le monde. Il est bien connu que la volaille, en particulier le poulet de chair, constitue le principal réservoir pour ce pathogène zoonotique. Par conséquent, la prévention et la surveillance de Salmonella au cours de la phase d'élevage pourrait réduire efficacement la contamination de la viande à l'abattoir et lors de la transformation. Avec l'interdiction de l'utilisation sub-thérapeutique des antibiotiques en Europe et la rigueur croissante de la législation européenne en matière d'hygiène alimentaire, l'immunisation passive par l'administration orale d'anticorps de jaune d'oeuf (IgY) spécifiques du pathogène serait une alternative intéressante. Cette synthèse bibliographique donne des informations concises sur la production des IgY et leur utilisation en immunisation passive, en particulier chez la volaille. Mots clés. Anticorps du jaune d'oeuf, production de l'IgY, immunisation passive, infections entériques, additif alimentaire.
The Production of Avian (Egg Yolk) Antibodies: IgY
Alternatives to Laboratory Animals, 1996
for the Validation of Alternative Methods. 2 This document represents the agreed report of the participants as individual scientists. 926 22-24 March 1996, under the chairmanship of Christian Staak (BgVV, Berlin, Germany). The participants at the workshop, who were representatives from academia, industry and government, reviewed the current status of immunoglobulin Y production in the chicken. Particular emphasis was placed on discussion of: a) the housing of chickens; b) immunisation protocols; c) egg-laying behaviour; d) the isolation and purification of lgY; and e) the successful replacement of the use of mammalian antibodies by IgY. The main issues discussed and recommendations made in connection with these topics are summarised in this report.
International Journal of Electrochemical Science, 2013
Chicken antibodies isolated from egg yolks (IgY) are a suitable alternative to conventional antibodies from blood of experimental animals. Main advantages of IgY are: (i) larger amounts of IgY could be prepared from repeatedly laid chicken eggs, (ii) collection of eggs is a non-invasive procedure contrary to bleeding of mammals, and (iii) a better response of chicken to mammalian antigens due to the larger evolutionary distance. Here, we report a cost-and labor-effective two-step procedure consisting of yolk extraction by tap water (8-fold dilution, freezing, and filtration) followed by a specific precipitation of IgY at pH 4 with sodium chloride in the final concentration of 8.8%. Using this procedure, the highly purified antibody (97%) was prepared. Resulting IgY preparations are acceptable for a wide range of applications as it was prepared using only bio-compatible chemicals. The purification protocol was developed and optimized in terms of the time, materials and the necessary manipulations.
Immunogenicity studies of various experimental vaccines in chickens
2020
In this paper, the main objective was to raise chickens' antibodies against three crucial public health microorganisms: the human immunodeficiency virus-1, Salmonella spp, and Staphylococcus aureus. Immunogens were prepared from the said microorganisms. Chickens were vaccinated either orally or intramuscularly. After a booster immunization, mostly eggs were collected and assess for the presence of specific antibodies. The most important results were the production of a large amount of anti-HIV antibodies in chicken's eggs, and also the synthesis of anti-protein A antibodies with the ability to inhibit the growth of S. aureus in vitro and to serve as anti-anti-idiotypic antibodies with the capacity of neutralizing the original antigen. Enzymelinked immune absorbent assays detected the presence of these antibodies as anti-Salmonella antibodies that were critical in reducing the bacterial load in the stomach and caeca compared with a control group. The vaccines were effective and safe, but more laboratory work, and economics have to be carried out to start a human trial.
Avian eggs present an ideal alternative antibody source to mammals as the immunoglobulin (IgY) in the chicken's blood is transported to the egg and accumulates in the egg yolk in large quantities. The existence of an immunoglobulin G (IgG)-like molecule in avian eggs, referred to as IgY, has been well documented, and extensive research has been carried out on its characterization, production and purification. Although it is the functional equivalent of mammalian IgG, the major serum antibody found in mammals IgY is structurally different, and has been found to exhibit several important differences when compared to mammalian antibodies, including its physicochemical properties and immunological capabilities. Recently, considerable research has focus seldom use of IgY as an alternative to mammalian antibodies for several applications, including immunotherapeutic applications, especially for the oral passive immunization against various bacteria and viruses. Much research has also been carried out on the use of IgY as a replacement for IgG in various immunodiagnostic and immunoaffinity purification purposes. The use of IgY offers several advantages over polyclonal antibodies produced in mammals, including providing a much more hygienic, cost efficient, convenient, humane and plentiful source of antigen-specific antibodies.