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IgY extraction and purification from chicken egg yolk

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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.


Extraction and purification; immunoglobulin; chicken egg yolk

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Akita EM, Nakai S (1992) Immunoglobulins from egg yolks: isolation and purification. J Food Sci 57:629-634.

Akita EM, Nakai S (1993) Comparison of four purification methods for the production of immunoglobulins from eggs laids hens immunized with enterotoxigenic E. coli strain. J Immunol Methods 160:207-214.

Al-Adwani SR, Crespo R, Shah DH (2013). Production and evaluation of chicken egg-yolk-derived antibodies against Campylobacter jejuni colonization-associated proteins. Foodborne Pathog Dis 10: 624-631.

Al-Edany AA (2011) Effect of immunoglobulin y purified from immunized hen eggs on the growth of Staphylococcus aureus. Bas J Vet Res 10:85-96.

Araújo AS, Lobato ZIP, Chávez-Olórtegui C, Velarge DT (2010) Brazilian IgY-Bothrops antivenom: Studies on the development of a process in chicken egg yolk. Toxicon 55: 739–744.

Asemota H, Curtello S, Vaillant AAJ, Mohammed W, Vuma S, Chalapathi Rao AV, Kurhade A, Kissoon S, Smikle M, Wisdom B, Kurhade G (2013) Purification of Avian IgY with trichloroacetic acid (TCA). J Chromatogr Separ Technique 4:205. doi:10.4172/2157-7064.1000205.

Bade H, Stegemann H (1984) Rapid method of extraction of antibodies from hen egg yolk. J Immunol Methods 72: 421-426.

Bizhanov G, Vyshniauskis G (2000) A comparison of three methods for extracting IgY from the egg yolk of hens immunized with Sendai virus. Vet Res Commun 24: 103-113.

Bižanov G, Jonauskienė I (2003) Production and purification of IgY from egg yolk after immunization of hens with pig IgG. B Vet I Pulawy 47:403-410.

Bizhanov G, Jonauskiene I, Hau J (2004) A novel method, based on lithium sulfate precipitation for purification of chicken egg yolk immunoglobulin Y, applied to immunospecific antibodies against Sendai virus. Scand J Lab Anim Sci 31:121-130.

Burley RW, Cook WH (1961) Isolation and composition of avian egg yolk granules and their constituent α and β-lipovitellins. Can J Biochem Physiol 39:1295-1307.

Carlander D (2002) Avian IgY antibody: in vitro and in vivo. PhD Thesis. University of Uppsala, Uppsala, Sweden.

Chang HM, Lu TC, Chen CC, Tu YY, Hwang JY (2000) Isolation of immunoglobulin from egg yolk by anionic polysaccharides. J Agr Food Chem 48:995-999.

Dai YC, Wang YY, Zhang XF, Tan M, Xia M, Wu XB, Jiang X, Nie J (2012) Evaluation of anti-norovirus IgY from egg yolk of chickens immunized with norovirus P particles. J Virol Methods 186:126-131.

Deignan T, Kelly J, Alwan A, O’Farrelly C (2000) Comparative Analysis of Methods of Purification of Egg Y Yolk Immunoglobulin. Food Agr Immunol 12:77–85.

De Meulenaer B, Baert K, Lanckriet H, Hoer VV, Huyghebaert A (2002) Development of an enzyme-linked immunosorbent assay for bisphenol A using chicken immunoglobulins. J Agr Food Chem 50:5273-5282.

Dubie T, Sisay T, Zeru F, Gebru M-u, Muktar Y (2015) The potential application of avian egg antibodies with emphasis on immunotherapeutic and immunodiagnostic purpose. J Vet Med

Anim Health 7: 145-158.

Hansen P, Scoble JA, Hanson B, Hoogeraad NJ (1998) Isolation and purification of immunoglobulins from chicken eggs using thiophilic chromatography. J Immunol Methods 215:1-7.

Hassl A, Aspöck H (1988) Purification of egg yolk immunoglobulins. A J HELLENIC VET MED SOC 2017, 68(3) ΠΕΚΕ 2017, 68(3) two-step procedure using hydrophobic interaction chromatography and gel filtration. J Immunol Methods 110:225-228.

Hatta H, Kim M, Yamamoto T (1990) A novel isolation method for hen egg yolk antibody, ‘IgY’. Agr Biol Chem 54:2531-2535.

Hernández-Campos FJ, Brito-De la Fuente E, Torrestiana-Sánchez B (2010) Purification of egg yolk Immunoglobulin (IgY) by ultrafiltration: effect of pH, ionic strength, and membrane properties. J Agr Food Chem 58:187–193.

Hodek P, Trefil P, Simunek J, Hudecek J, Stiborova M (2013) Optimized protocol of chicken antibody (IgY) purification providing electrophoretically homogenous preparations. Int J

Electrochem Sci 8:113–124.

Jensenius JC, Andersen I, Hau J, Crone M, Koch C (1981) Eggs: conveniently packaged antibodies. Methods for purification of yolk IgG. J Immunol Methods 46:63-68.

Jensenius JC, Koch C (1993) On the purification of IgG from egg yolk. J Immunol Methods 164:141-142.

Júnior ÁF, Santiago FM, Silva MV, Fereira FB, Júnior AGM, Mota CM, Faria MS, Siva Filho HH, Silva DAO, Cunha-Júnior JP, Mineo JR, Mineo TWP (2012) Production, characterization and applications for Toxoplasma gondii-specific polyclonal chicken egg yolk immunoglobulins. PLoS One 7:1-10.

Kassim N, Mtenga AB, Shim WB, Chung DH (2012) The in vitro and in vivo efficacy of hen IgY against Vibrio parahaemolyticus and Vibrio vulnificus. J Microbiol Biotechnol 22:1423–1431.

Kates M (1972) Techniques of lipidology. Isolation, analysis and identification of lipids. North-Holland Publishing, Amsterdam. Kim H, Nakai S (1996) Immunoglobulin separation from egg yolk: a serial filtration system. J Food Sci 61:510-512.

Kim H, Nakai S (1998) Simple separation of immunoglobulin from egg yolk by ultrafiltration. J Food Sci 63:485-490.

Kitaguchi K, Minoura M, Noritake M, Mizutani M, Kinoshita K, Horio F, Murai A (2008) Determination of immunoglobuluin Y concentration in yolk extract prepared by water dilution method: comparisons among three strains of chickens. J Poult Sci 45:82-87.

Ko KY, Ahn DU (2007) Preparation of immunoglobulin Y from egg yolk using ammonium sulfate precipitation and ion exchange chromatography. Poultry Sci 86:400-407.

Kovacs-Nolan J, Phillips M, Mine Y (2005) Advances in the value of eggs and egg components for human health. J Agr Food Chem 53:8421-8431.

Kwan L, Li-Chan N, Helbig N, Nakai S (1991) Fractionation of water soluble and insoluble components from yolk with minimum use of organic solvents. J Food Sci 56:1537-1541.

Liou JF, Shiau JW, Tai C, Chen LR (2011) Production of egg yolk immunoglobulin against Escherichia coli from white Leghorn and Lohmann chickens. J Anim Vet Adv 10:2349-2356.

Mahdavi AH, Rahmani HR, Nili N, Samie AH, Soleimanian-Zad S (2010) Egg yolk antibody (IgY) powder against Escherihia coli O78:K80. J Anim Vet Advan 9:366-373.

Marcet I, Laca A, Paredes B, Díaz M (2011) IgY isolation from a watery by-product obtained from an egg yolk fractionation process. Food Bioprod Process 89:87-91.

McCannel AA, Nakai S (1990) Separation of yolk immunoglobulins into subpopulations using DEAE-ion exchange chromatography. Can Inst Food Sci Technol J 23:42-46.

McCully KA, Mok CC, Common RH (1962) Paper electrophoretic characterization of proteins and lipoproteins of hen’s egg yolk. Can J Biochem Physiol 40:937-952.

Nafea NM (2012) Isolation, purification and conjugation of polyclonal antibodies type Y from immunized hens against Hepatitis B surface antigen. MS Thesis. University of Baghdad, Baghdad, Iraq.

Ntakarutimana V, Demedts P, Van Sande M, Scharpé S (1992) A simple and economical strategy for downstream processing of specific antibodies to human transferrin from egg yolk. J Immunol Methods 153:133-140.

Parma YR, Chacana PA, Lucchesi PM, Roge A, Granobles Velandia CV, Krüger A, Parma AE, Fernández-Miyakawa ME (2012) Detection of Shiga toxin-producing Escherichia coli by sandwich enzyme linked immunosorbent assay using chicken egg yolk IgY antibodies. Front Cell Infect Microbiol 2:1-8.

Polson A, Von Wechmar MB, Van Regenmortel MVH (1980a) Isolation of viral IgY antibodies from yolks of immunized hens. Immunol Commun 9:475-493.

Polson A, Von Wechmar MB, Fazakerley G (1980b) Antibodies to proteins from yolk immunized hens. Immunol Commun 9:495-514.

Polson A, Coetzer T, Kruger J, Von Maltzahn E, Van der Merwe KJ (1985) Improvements in the isolation of IgY from the yolks of eggs laid by immunized hens. Immunol Invest 14:323-327.

Ruan GP, Ma L, He XW, Meng MJ, Zhu Y, Zhou MQ, Hu ZM, Wang XN (2005) Efficient production, purification, and application of egg yolk antibodies against human HLA-A*0201 heavy chain and light chain (β2m). Protein Expres Purif 44:45-51.

Russel WMS, Burch RL (1959) The principles of humane experimental technique. Methuen, London.

Sampaio LC, Baldissera MD, Grando TH, Gressler LT, Capeleto DdeM, de Sa MF, de Jesus FP, dos Santos AG Jr, Anciuti AN, Colonetti K, Stainki DR, Monteiro SG. (2014). Production, purification and therapeutic potential of egg yolk antibodies for treating Trypanosoma evansi infection. Vet Parasitol 204:96-103.

Shahbazi P, Shayan P, Ebrahimzadeh E, Rahbari S (2009) Specific egg yolk antibody against recombinant Cryptosporidium parvum P23 protein. Iran J Parasitol 4:15-24.

Shenstone FS (1968) The gross composition, chemistry and physicochemical basis of organization of the yolk and white. In: (ed.: Carter TC) Egg quality a study of hen’s egg, Oliver and Boyd, Edinburgh, pp.26-58.

Sudjarwo SA, Sudjarwo KE, Koerniasari (2012) Purification and characterization protein of anti-dengue specific immunoglobulin Y for diagnostic kit of denge. J Appl Pharm Sci 2:7-12.

Sunwoo HH, Lee EN, Gujral N, Suresh MR (2010) Growth inhibition of Escherichia coli 987P by neutralizing IgY antibodies. Open Immunol J 3:1-8.

Verdoliva A, Basile G, Fassina G (2000) Affinity purification of immunoglobulins from chicken egg yolk using a new synthetic ligand. J Chrom B 749:233-242.

Vieira JG, Oliveira MA, Maciel RM, Mesquita CH, Russo EM (1986) Development of an radioimmunoassay for the synthetic amino terminal (1-34) fragment of human parathyroid hormone using egg yolk-obtained antibodies. J Immunoassay 7:57-72.

Vega C, Bok M, Chacana P, Saif L, Fernandez F, Parreño V (2011) Egg yolk IgY: protection against rotavirus induced diarrhea and modulatory effect on the systemic and mucosal antibody responses in newborn calves. Vet Immunol Immunopathol. 142: 156–169.

Wen J, Zhao S, He D, Yang Y, Li Y, Zhu S (2012) Preparation and characterization of egg yolk immunoglobulin Y specific to influenza B virus. Antiviral Res 93:154-159.

Williams J (1962) Serum proteins and the livetins in hen’s-egg yolk. Biochem J 83:346-355.

Xu JJ, Ren CZ, Wang SS, Liu DD, Cao LQ, Tao JP (2013) Protection efficacy of multivalent egg yolk immunoglobulin against Eimeria tenella infection in chickens. Iran J Parasitol 8:449-458.


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