Chicken Egg Yolk Antibodies, Production and Application: IgY-Technology ( Springer Lab Manuals): Medicine & Health Science Books. Chicken egg yolk antibodies (IgY-technology): a review of progress in The production of antibodies (Abs) in chickens and the extraction of specific Abs from used IgY-extraction procedures; g) new possibilities for application in human and. Production and purification of IgY antibodies from chicken egg yolk that IgY technology can be used for the production of primary antibodies for immunological egg yolk to achieve high yield of production for research and commercial uses.

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September 22nd Reviewed: February 20th Published: Immunized chicken transfers a amtibodies IgY from blood to egg yolk. Phylogenetic distance between chickens and mammals influences the generation of antibody repertoires recognizing an antigen profile. Toxoplasma gondii is the toxoplasmosis agent in human and animals and is a worldwide public health problem. Alternative approach that offers possibility to improve diagnostics, treatment, or control of toxoplasmosis should be accepted as a protective approach to hosts at risk of infection by T.

Hens transfer IgY from blood to egg yolk protecting their offspring. Isolation of yolk IgY avoids bleeding the chicken and reduces the number of painful procedures. Hens can lay approximately one egg each day during a period of 2 years.

IgY antibody is functional and is equivalent to mammal IgG antibody. Both avian and mammalian antibodies have structure similarity. They have two aplication light chains and two identical heavy chains bounded to each other by disulfide bonds. In the carboxy terminal region, they have the fragment crystallizable Fc that is associated to biological antibody functions as interaction with cells or molecules from immune system.

Virtually, any substance can be the target of an antibody response and the response to even a single epitope comprises many different antibody molecules, each with a subtly different specificity for the epitope and a unique affinity, or binding strength. The total number of antibody specificities available for an individual is known tecnology the antibody repertoire.

Albeit, they are equivalent in function and structure, the phylogenetic distance and the way chicjen generate antibody specificities produce difference in epitope recognition repertoire between avian and mammalian host. Additionally, modifications in amino acid sequence of IgY antibody result in advantages, comparing to IgG, as follows: Purified IgY antibody can be applied to several kinds of techniques including screening new antigens by phage display, enzymatic or florescence immunoassays, diagnostics, and, also, passive immunization for therapy against gastrointestinal pathogens in humans and animal hosts.

In addition, create new diagnostic platforms for diagnosing T. Finally, the potential protective effect of IgY to be used in prevention or toxoplasmosis therapy by passive immunization should be explored.

In the earlyKlemperer published his observation that there must be neutralizing proteins i. In this context, phylogenetic distance between chickens and mammals, mechanisms of antibody repertoire diversification, and the way in which chickens deposit IgY immunoglobulin in the egg yolk provide a number of advantages compared to mammals as animal model for antibody production.

The generation and application of avian antibodies has caused a surge of interest in a wide variety of applications within the life sciences [ 1 ]. For both light chain and heavy chain, variable domains VL and VH have three regions with higher variation rate in amino acid sequence named complementarity determining region CDR. Complementarity determining regions CDRs are completely bounded to epitopes antigenic determinant during a specific humoral response.

Prouction the direction to antiobdies terminal extremity, after the variable domain, there are the constant domains. The light chain has one constant domain CL ; however, the number of constant domains for the heavy chain varies with antibody isotypes. There was proposed that IgY CH2 domain must be converted to IgG hinge region, which provides a higher mobility to tehcnology mammalian Fab than to its avian equivalent. To generate diversity, mammals depend on combinatorial and functional variations that occur during the gene rearrangement events to produce complete heavy and light chain Ig genes.

This gene rearrangement process goes continuously in the bone marrow, where each developing B cell assembles a unique heavy and light chain Ig gene from families of functional V variableD diversityand J joining gene segments.

In contrast, chickens have only single functional V and J segments for the heavy and light chain loci, and chicken Ig gene rearrangement occurs only during a brief period of embryonic development.

Within the bursa, B cells also acquire somatic diversity among the rearranged V gene segments of the heavy and light chain Ig loci. Somatic diversification of chicken V gene segments occurs by intrachromosomal gene conversion, a DNA recombination process which involves unidirectional transfer of nucleotide sequence blocks from the families of V region pseudogenes into the functional rearranged VH and VL genes [ appplication ].


IgY is the predominant isotype in sera, produced after IgM in the primary antibody response, and it is the main isotype produced in the secondary immune response. IgY has different biochemical properties from those of mammalian IgG antibodies and shares homology while functioning with them.

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Due to the lack of the hinge region, IgY has limited flexibility to its Fab resulting in to precipitate antigens at physiological salt concentrations.

It has a MW of about kDa. IgY sampling is not invasive, which confers advantage on IgG; also, yolk antibody does not interfere with rheumatoid factor or activate mammalian complement [ 1 ]. First step is the chicken immunization followed by the yolk IgY extraction, the antibody characterization, and finally IgY applications in various kinds of assays. Chickens are a suitable model to produce IgY by immunization with nucleic acid [ 6 ], protein [ 7 ], lipid, and carbohydrates [ 8 ].

In addition, an immune humoral response must be elicited by immunization with recombinant proteins chciken 9 ] or peptides [ 10 ]. Intramuscularly way is a common way for chicken immunization to produce specific IgY. Alternatively, subcutaneous way provides satisfactory amounts of specific antibody [ 1 ]. General layout to produce specific IgY antibodies. From immunization with different antigens added to antibdoies, isolating egg yolk antibody to IgY applications.

The concentrations of specific antibodies in the yolk are similar to the antibody profile of the serum. To isolate yolk IgY, distinct procedures may be used alone or in combination according to the following criteria: The yolk antibody extraction begins by separating yolk and egg anr. After removal of the vitellin membrane, the yolk is put into a measuring cylinder.

The next steps in IgY extraction are devoted to removing the lipid content and obtaining a water soluble phase WSP antibodies enriched [ 12 ]. The consequent purification procedures vary widely in type and combination and from laboratory to laboratory.

The degree of IgY purity depends on desired application; for example, for oral administration, the whole yolk can be used, and on the other hand, for a reagent, isolated IgY from the mixture of yolk molecules is necessary [ 1 ]. A high quality removing lipid method is the first step to efficiently isolate IgY antibodies from egg yolk.

Optionally, organic solvents chloroform have been used as a mean to remove yolk lipid content at the initial step of IgY isolation [ 114 ]. The extraction of yolk antibodies by the use applicahion polyethylene glycol precipitation PEG, MW was, firstly, developed by Polson et al.

Afterwards, PEG was widely accepted as a standard procedure. Since the precipitation of certain proteins from a mixture by NaS or AmS depends on the concentration of the salt, this procedure can be used not only for isolation of desired proteins but also for elimination of the undesired ones.

A precipitate is obtained through centrifuging, taken up in a certain amount of desired buffer or distilled water, and finally dialyzed proeuction 1 ]. NaS and AmS precipitations are economical and environmentally correct procedures for yolk antibody extraction [ 24 ].

NaS or AmS precipitation provides fractions with low protein content compared to original yolk, but with a high specific activity [ 26 ]. The isolation of IgY from yolk using of combination of different methods results in a high amount of specific antibodies with elevated purity. Association between AmS and ethanol to IgY precipitation results in a high quantity of antibodies with great purity [ 31 ].

Specific antisnake venom IgY with neutralizing effect for therapeutic proposes can be isolated by AmS applicztion followed to ion exchange chromatography [ 3233 ]. Similarly, growth inhibition of Staphylococcus aureus and Escherichia coli was obtained with yolk antibodies isolated by NaS and AmS precipitations [ 35 ]. The separation of IgY antibodies according to their molecular weight through gel filtration is a further stage of purification of yolk antibodies but rarely reported except in the desalting of preparation through columns instead through a dialysis [ 1 ].

According to the strength of binding, the antibodies are detached by increasing concentrations of ions of the buffers [ 1323334 ]. The isolation of yolk IgY antibodies by affinity chromatography consist in attach an antibody ligand on a matrix.

IgY antibodies bind neither with protein A nor with protein G [ 1 ]. Recently, protein M a transmembrane protein from human mycoplasma has been demonstrated as a promising ligand for purifying polyclonal, monoclonal, or engineered IgY antibodies [ 38 ]. IgY antibodies are suitable for a wide application protocols like proteomic, diagnostics, and therapy. Other possibility for the use of IgY igh proteomic studies is screening random peptide phage display library to detect new epitope candidates to be applied for diagnostic methods to widespread parasitic human diseases [ 46 ].


Chicken Egg Yolk Antibodies, Production and Application : IgY-technology

Appliation focused on a number of defined targets can be constructed using lymphocyte mRNA from chickens immunized with a mixture of several different antigens from Plasmodium falciparum, Trypanosoma sp. Relevant to consider that antigenic targets recognition by the use of antibodies can show some differences according to the method used. Regarding the conformational epitope structure, the performance of antibodies recognizing a given epitope by WB can represent nothing about the same antibody performance in ELISA with the same antigen [ 49 ].

IgY antibody shows advantages comparing to the mammal IgG use in immunoassay. Egg yolk IgY can be labeled with horseradish peroxidase for use in immunoenzymatic assays [ 52 ]. IgY antibodies can be applied to immunofluorescence and flow cytometry assays. Chicken IgY and rabbit IgG conjugations with fluorescein isothiocyanate FITC comparatively used to examine strains of Campylobacter fetus revealed that both conjugates have a high percentage rate of detection; IgY has less background due to unspecific fluorescence than IgG.

Additionally, IgY is a cheap, bloodless, and very productive method [ 63 ]. IgY antibodies are suitable reagents for flow cytometry and as well as certain monoclonal antibodies, for example, to study human and rabbit platelet physiology [ 66 ]. The immunochromatographic assay ICA requires no instruments and has a cicken time of less than 10 minutes and it is portable and easy to perform in the field.

The passive immunization is a protective method, which has been tested for many years and shown to be effective [ 71 ]. The use of IgY offers several advantages such as it is environmentally friendly, nontoxic, reduces the numbers of animals required for antibody production, stability in the orogastrointestinal tract, and its safety profile [ 71 — 74 ].

A major obstacle to its implementation is its relatively high cost, which is dependent, among other things, mainly on two factors: Regarding the IgY concentration in egg yolk and blood of laying hens, a recent study provides evidence that there is a significant circaseptan rhythm in yolk IgY and circaquattran rhythm in serum IgY.

Additionally, the serum IgY concentration reached to maximum in the morning, decreased to minimum during the daytime, and increased again at night revealing a significant circadian rhythm, which may reflect in yolk antibody concentration [ yoolk ].

The microencapsulation with a methacrylic acid copolymer may be an effective method of protecting purified yolk IgY from applicatoon inactivation, enabling its use for oral passive immunotherapy [ 77 ]. The use of chitosan-alginate microcapsules to protect IgY from gastrointestinal environment conditions provides significant resistance to pepsin hydrolysis and may enable intact IgY to reach target chiclen within the lower digestive tract [ 78 ].

Another approach protecting IgY from degradation in gastric pH can be the incorporation of antibodies to hydrogel containing acrylamide and acrylic acid with promising results for IgY oral delivery [ 79 ]. Oral administration of IgY antibodies has been tested for many years with promising results [ 80 ] to different pathogens as human rotavirus [ 81 ]; dental plaque formation by Streptococcus mutans [ 8283 ]; enteropathogenic E.

Chicken Egg Yolk Antibodies, Production and Application : Reudiger Schade :

Immunotherapy as a appoication immunization method to neutralize venom using purified IgY proved to be efficient for therapy protocol [ — ]. Currently, there are a limited number of studies about IgY antibody production against T. They used total soluble antigens that are immunogenic to comparatively produce rabbit IgG and chicken IgY. The results indicated differences between the specificities of egg yolk IgY antibodies and rabbit IgG serum antibodies, although both animal species had been immunized with identical antigen preparations.

Ferreira Junior et al.

They purified yolk antibodies by water dilution method, sodium sulfate precipitation, and molecular weight exclusion chromatography.

Specific antibody characterization was due to both indirect ELISA and Western blotting methods and applied to immunofluorescence assays by immunohistochemistry and immunocytochemistry assays. Using the Western blotting method, the antigenic bands recognition profile were different between mice and chicken antibodies.

IgY antibodies detected parasites in cytoplasm of infected cells and in brain samples of infected mice.