Affinity purification of immunoglobulins from chicken egg yolk using a new synthetic ligand

Verdoliva, Antonio; Basile, Giancarlo; Fassina, Giorgio

doi:10.1016/s0378-4347(00)00426-6

Cited by 82 publications

(52 citation statements)

References 33 publications

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“…These techniques include the following methods, such as anion exchange chromatography (Bade and Stegemann, 1984), affinity chromatography (Ntakarutimana et al, 1992), thiophilic chromatography (Hansen et al, 1998), or ion exchange chromatography (Ko and Ahn, 2007). As a result, the yield of IgY was not more than 1 mg/ml of egg yolk (Hassl and Aspöck, 1988;McCannel and Nakai, 1990;Verdoliva et al, 2000). Furthermore, it was reported that purity of IgY obtained by cation exchange chromatography was not high and contained too many contaminating proteins.…”

Section: Purification Og Igymentioning

confidence: 99%

IgY extraction and purification from chicken egg yolk

Bižanov

2018

J Hellenic Vet Med Soc

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ABSTRACT.The interest for immunoglobulin Y (IgY) isolation arises from the possible applications of these immunoglobulins in diagnostics and therapeutics. Powered whole eggs or yolks have been used in veterinary medicine as an inexpensive immunoglobulin Y source for the treatment of enteric diseases. Incorporating feed grade egg yolk antibodies into animal diets has been examined extensively in an attempt to limit pathogenic diarrhea causing by Escherichia coli in swine, and limit Salmonella establishment in calves and mice, as well as Campylobacter, Clostridium, and Salmonella in poultry. Thus, therapeutic administration of the IgY might reduce the clinical use of antibiotics, and minimize the risk of developing bacterial antibiotic resistance. Methods of immunoglobulins isolation and purification from hen yolk are reviewed. For a large-scale production, one of the problems is separating the water-soluble protein fraction from the lipids and other hydrophobic substances. Precipitation or aggregation of lipid occurs under various conditions: (1) by use of water dilution under acidic conditions; (2) by means of acids; (3) combination of solvents (chloroform, aceton) which selectively solubilize the lipids; (4) using of 3.5% polyethylene glycol and (5) by applying natural gums (polyanionic polysaccharides). Further purification of IgY after crude extraction can be achieved by selective precipitation. IgY precipitation can be carried out by salt precipitation using saturated solutions of sodium or ammonium sulphate, which dehydrate proteins. Furthermore, selective precipitation is achieved by using 8.8% sodium chloride or at 12% of polyethylene glycol. The methods comparison on the basis of IgY yield, showed that the mean IgY yield obtained by water dilution method or precipitation with salts, or by using polyethylene glycol was 5.6 mg/ml of egg yolk, 6.3 mg/ml of egg yolk and 8.7 mg/ml of egg yolk correspondingly and did not significantly differ between them. Water dilution method seemed to offer the best IgY recoveries. Moreover, this method in combination with chromatography and filtration can be applied easily in an industrial environment. Filtration technology offers the best opportunities for industrial applications while precipitation with polyethylene glycol or salts provides a cheap and easy methodology for laboratory use. In the production of safer foods, an important strategy is to exploit natural antimicrobial agents as alternatives to conventional synthetic chemical preservatives. In this regard, the IgY is of much interest for its potential application in fortified foods, such as administration for prevention of enteric diseases. The use of IgY is cost-effective.

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Section: Purification Og Igymentioning

confidence: 99%

IgY extraction and purification from chicken egg yolk

Bižanov

2018

J Hellenic Vet Med Soc

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“…Firstly, simple water dilution method is used to remove lipoproteins from WSF for the IgY purification (Jensenius et al, 1981;Kwan et al, 1991;Akita & Nakai, 1992;1993). This water dilution method has demonstrated as the best method to recover and purify the IgY with the highest purity of 34% in comparison with the methods of polyethylene glycol, dextran sulphate and chloroform (Verdoliva et al, 2000). However, the purity (74-78%) of the IgY from WSF with water dilution method was lower than that (83-99%) of the IgY from WSF with filter paper (Kim & Nakai, 1998).…”

Section: Resultsmentioning

confidence: 99%

“…Generally, affinity chromatography and ion exchange chromatography are the best methods to purify the IgY. Verdoliva et al (2000) obtained the highest yield of the IgY (10.2 mg/mL) and the best purity (> 90%) from the WSF using TG19318/Emphaze and TG19318/Emphaze affinity column. Additionally, affinity chromatography with synthetic ligand could gain the 92.1% purity and the 78.2% recovery of the IgY (Dong et al, 2008).…”

Section: Resultsmentioning

confidence: 99%

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Extraction and Purification of Anti-Helicobacter pylori IgY

Siriya¹,

Chu²,

Chen³

et al. 2013

JAS

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The aim of this study was to establish an efficient procedure to produce anti-H. pylori urease IgY from the eggs of the laying hens. Four White Leghorn hens (52 weeks old) were immunized intramuscularly with the purified urease of H. pylori with aluminum hydroxide gel adjuvant. To increase the specificity and antibody titer, three boosters were given at 2-week intervals following the first injection. After the final immunization, the eggs were collected daily and stored at 4°C. The yield of yolk proteins were 47.52%, 41.07% and 51.35% for the methods of dextran sulphate, isopropanol and water-soluble fraction (WSF), respectively. Further, IgY was purified successively by ammonia sulfate, affinity chromatography and ion exchange chromatography for each crude protein. Ammonia sulfate salting produced the IgY with a purity of 37-53% and the titer of 22.6-29.5 unit/mg. The purity, relative purification folds and titer characteristics of the purified IgY were 85-90%, 19.63-34.61 and 223.8-273.4 unit/mg protein for affinity chromatography, and 89-92%, 25.33-47.14 and 288.8-372.4 unit/mg protein for ion exchange chromatography, respectively. This anti-Helicobacter pylori IgY was specific against H. pylori usease. Here, we established a simple and inexpensive method to obtain a yield of 90% purity of the anti-H. pylori IgY in combinations of the WSF method, ammonia sulfate precipitation, and affinity chromatography.

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“…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]. Synthetic IgY-ligands were developed to improve yolk antibody isolating mainly by reducing the purification steps and increasing the amount and purity of recovered IgY [39,40]. Attaching the IgY-induced antigen to matrix provides another way to extract a high-purity IgY by affinity chromatography [41,42].…”

Section: Isolating Igy By Affinity Chromatographymentioning

confidence: 99%

IgY-Technology Applied to Studies of Toxoplasma gondii Infection

Júnior¹,

Santos²,

Bassi³

et al. 2017

Toxoplasmosis

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In this chapter, we describe relevant aspects of immunoglobulin Y (IgY) technology for Toxoplasma gondii applications, including comparison of avian IgY antibody with mammalian IgG antibody, egg yolk IgY production and isolation procedures, important applications for IgY antibody, and state of the art and perspectives for IgY-technology in T. gondii studies. T. gondii is a worldwide public health problem. IgY-technology provides an alternative antibody (IgY) to mammalian Immunoglobulin G (IgG) antibody. IgY-technology involves the chicken immunization, yolk IgY isolation, antibody characterization, and purified IgY application to several kinds of methods. Immunized chicken transfers a specific IgY from blood to egg yolk. Phylogenetic distance between chickens and mammals influences the generation of antibody repertoires recognizing an antigen profile. IgY is not bound to rheumatoid factor or mammalian complement protein and thus avoids the false-positive results. Yolk IgY isolation is carried out by simple procedures that are accessible for any laboratory and, also, for IgY isolation at large-scale production. IgY-technology provides antibodies for proteomic studies, diagnostic assays, and immunotherapy. Although IgY-technology is promising, there is a reduced number of investigations with IgY and T. gondii. Future perspectives involve the use of IgY-technology for the screening of new T. gondii antigens for diagnostics, therapy, or vaccine, development of innovative techniques for toxoplasmosis diagnostics and may be an immunotherapy for toxoplasmosis.

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Affinity purification of immunoglobulins from chicken egg yolk using a new synthetic ligand

Cited by 82 publications

References 33 publications

IgY extraction and purification from chicken egg yolk

IgY extraction and purification from chicken egg yolk

Extraction and Purification of Anti-Helicobacter pylori IgY

IgY-Technology Applied to Studies of Toxoplasma gondii Infection

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