Investigation of Two Precipitation Methods for Extracting Immunoglobulin Y (IgY) from Egg Yolks
DOI:
https://doi.org/10.30564/vsr.v3i2.4074Abstract
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) by using chloroform precipitate. The 2nd week post-immunization, IgY concentration increased respectively to 3903 ± 726 μg.ml-1 (chloroform extraction process) and 2937 ± 294 μg.ml-1 (PEG extraction process) (P < 0.01). After 3rdimmunization, IgY level obtaining from in immunization group extracted by chloroform process (6633 ± 1166 μg.ml-1) increased 2.7 times higher than that in control group (2482 ± 414 μg.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 SDSPAGE. 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.
Keywords:
Immunoglobulin Y; Egg yolk; Precipitation; Polyethylene glycol; ChloroformReferences
[1] Akita, E.M., Nakai, S., 1993. Comparson four purification methods for the production of immunoglobulins from egg laid by hens immunized with an enterotoxigenic E. coli strain. J Immunol Methods. 160(2), 207-214.
[2] Al-Adwani, S.R., Crespo, R., Shah, D.H., 2013. Production and evaluation of chicken egg-yolk derived antibodies against Campylobacter jejuni colonization-associated proteins. Foodborne Pathog Dis. 7(10), 624-631.
[3] Al-Edany, A., 2013. Preparation and Purification of Immunoglobulin Y from Egg Yolk of Immunized Hens. Journal of Thi-Qar University. 2(8), 7-16.
[4] Amro, W.A., Al-Qaisi, W., Al-Razem, F., 2018. Production and purification of IgY antibodies from
[5] chicken egg yolk. J Genet Eng Biotechnol. 16, 99- 103.
[6] 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 immune specific antibodies against Sendai virus. Scand J Lab Anim Sci. 31(3), 121-130.
[7] Bizanov, G., 2017. IgY extraction and purification from chicken egg yolk. J Hell Vet Medical Soc. 68, 265-272.
[8] Cakir-Koc, R., 2016. Production of anti-SAG-1 IgY antibody against Toxoplasma gondii parasites and evaluation of antibody activity by ELISA method. Parasitol Res. 115(8), 2947-2952.
[9] Carlander, D., 2002. Avian IgY Antibody: In vitro and in vivo. Acta Universitatis Upsaliensis. Comprehensive Summaries of Uppsala Dissertations from the Faculty of Medicine 1119, 53 pp. Uppsala. ISBN 91-554-5227-2.
[10] De Meulenaer, B., Baert, K., Lanckriet, H., Van Hoed, V., Huyghebaert, A., 2002. Development of an enzyme-linked immunosorbent assay for bisphenol A using chicken immunoglobulins. J Agr Food Chem. 50, 5273-5282.
[11] Horie, K., Horie, N., Abdou, A.M., Yang, J.O., Yun, S.S., Chun, H.N., Park, C.K., Kim, M., Hatta, H., 2004. Suppressive effect of functional drinking yogurt containing specifific egg yolk immunoglobulin on Helicobacter pylori in humans. J Dairy Sci. 87(12), 4073-4079.
[12] Hussain, C.N.B., 2017. Isolation and Estimation of Chicken Immunoglobulins (IgY) from Egg Yolk by Optimizing Polyethylene Glycol (PEG) Precipitation Method. Sch J Agric Vet Sci. 4(7), 286-292.
[13] Kim, H., Nakai, S., 1998. Simple separation of immunoglobulin from egg yolk by ultrafiltration. J Food Sci. 63, 485-490.
[14] Kumaran, T., Citarasu, T., 2016. Igy-technology: production of antibodies in chickens and use in therapy of infectious diseases. IJSRME. 1(1), 29-35.
[15] Lee, D.H., Jeon, Y.S., Park, C.K., Kim, S., Lee, D.S., Lee, C., 2015. Immunoprophylactic effect of chicken egg yolk antibody (IgY) against a recombinant S1 domain of the porcine epidemic diarrhea vírus spike protein in piglets. Arch Virol. 160(9), 2197-2207.
[16] Li, X., Yao, Y., Wang, X., Zhen, Y., Thacker, P.A., Wang, L., Shi, M., Zhao, J., Zong, Y., Wang, N., Xu, Y., 2016. Chicken egg yolk antibodies (IgY) modulate the intestinal mucosal immune response in a mouse model of Salmonella typhimurium infection. Int Immunopharmacol. 36, 305-314.
[17] Nasiri, K., Nassiri, M.R., Tahmoorespur, M., Haghparast, A., Zibaee, S., 2016. Production and characterization of egg yolk antibody (IgY) against recombinant VP8-S2 antigen. Pol J Vet Sci. 19(2), 271-279.
[18] Pauly, D., Chacana, P.A., Calzado, E.G., Brembs, B., Schade, R., 2011. IgY technology: extraction of chicken antibodies from egg yolk by polyethylene glycol (PEG) precipitation. J Vis Exp. 1(51), 1-6.
[19] Pizarro-Guajardo, M., Díaz-González, F., Álvarez-Lobos, M., Paredes-Sabja, D., 2017. Characterization of chicken IgY specifific to Clostridium difficile R20291 spores and the effect of oral administration in mouse models of initiation and recurrent disease. Front Cell Infect Microbiol. 7, 1-16.
[20] Polson, A., 1990. Isolation of IgY from the yolks of eggs by a chloroform Polyethylene glycol procedure. Immunol Commun. 19(3), 253-258.
[21] Polson, A., Wechmar, M.B., Fazakerley, G., 1980. Antibodies to proteins from yolk of immunized hens. Immunol Commun. 9(5), 495-514.
[22] Rahman, S., Nguyen, S.V., Faustino, C., Icatlo, Jr., Umeda, K., Kodama, Y., 2013. Oral passive IgY based immunotherapeutics: a novel solution for prevention and treatment of alimentary tract diseases. Hum Vaccin Immunother. 9(5), 1039-1048.
[23] Schade, R., 2001. Chicken Egg Yolk Antibodies, Production and Application IgY-Technology, Springer Berlin.
[24] Schade, R., Staak, C., Hendriksen, C., Erhard, M., Hugl, H., Larsson, G.K., Polimann, W., Van Regenmortel, M., Rijke, E., Spieimann, H., Steinbusch, H., Straughan, D., 1996. The production of avian (egg yolk) antibodies: IgY. The report and recommenda-30 Veterinary Science Research | Volume 03 | Issue 02 | December 2021 tions of ECVAM workshop 21, ATLA. 24, 925-934.
[25] Sunwoo, H.H., Nakano Dixon, T.W., Sim, J.S., 1996. Immune responses in chickens against lipopolysaccharide of Escherichia coli and Salmonella typhimurium. Poultry Sci. 75, 342-345.
[26] Suzuki, H., Nomura, S., Masaoka, T., Goshima, H., Kamata, N., Kodama, Y., Ishii, H., Kitajima, M., Nomoto, K., Hibi, T., 2004. Effect of dietary anti-Helicobacter pylori-urease immunoglobulin Y on Helicobacter pylori infection. Aliment Pharmacol Ther. 20, 185-192.
[27] Vega, C., Bok, M., Saif, L., Fernandez, F., Parreño, V., 12015. Egg yolk IgY antibodies: a therapeutic intervention against group a rotavirus in calves. Res Vet Sci. 103, 1-10.
[28] Walczak, M., Grzywa, R., Łupicka-Słowik, A., Skoreński, M., Bobrek, K., Nowak, D., Boivin, S., Eric, L., Brown, E.L., Oleksyszyn, J., Sieńczyk, M., 2015. Method for generation of peptide-specifific IgY antibodies directed to Staphylococcus aureus extracellular fifibrinogen binding protein epitope. Biopolymers. 104(5), 552-559.
[29] Yamamoto, H., Watanabe, H., Sato, G., 1975. Identification of immunoglobulins in chicken eggs and their antibody activity. Jap J Vet Res. 23, 131-40.
[30] Zhang, X., Diraviyam, T., Li, X., Yao, G., Michael, A., 2016. Preparation of chicken IgY against recombinant E2 protein of bovine viral diarrhea virus (BVDV) and development of ELISA and ICA
[31] for BVDV detection. Biosci Biotechnol Biochem. 80(12), 2467-2472.
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