Evaluation of Human Anti IgG Polyclonal Antibody Production Conjugated with Peroxidase in Egg Yolk
Background: Egg yolk is a rich and accessible source of yolk immunoglobulin (Y immunoglobulin). Presently, polyclonal antibodies from mammalian sources are used for diagnosis. Antibody production from egg yolk gives a higher yield and turnover than that from lab animals, and invasive methods such as phlebotomy and causing stress to the animals are not required. Due to the issues regarding mammalian antibodies, we aimed to evaluate the human anti-IgG polyclonal antibody production conjugated with peroxidase in egg yolk.
Methods: Population of laying hens reared in Agriculture/Isfahan University of Technology were used in 2017. After immunizing hen against pure human IgG, specific IgY (yolk immunoglobulin) was purified from the yolk by sedimentation with polyethylene glycol (PEG6000). To assess the molecular weight and activity of the product, SDS-PAGE and ELISA-test were used, respectively.
Results: The complete molecular weight of IgY was 180 kDa and the molecular weight of its light and heavy chains were 27 and 67 kDa, respectively.
Conclusion: Antihuman IgG IgY had a purity above 90%. The product of this study can be used to measure IgG class antibodies in order to diagnose different diseases.
2. Vidarsson G, Dekkers G, Rispens T (2014). IgG subclasses and allotypes: from struc-ture to effector functions. Front Immunol, 5: 520.
3. Gaetani G, Ambrosi E, Ugo PM, et al (2017). Electrochemical Immunosensor for Detection of IgY in Food and Food Supplements. Chemosensors, 5(1):10.
4. Melnik E, Bruck R, Muellner P, et al (2016). Human IgG detection in serum on pol-ymer based Mach-Zehnder interferomet-ric biosensors. J Biophotonics, 9(3): 218-223.
5. Agarwal S, Cunningham-Rundles C (2007). Assessment and clinical interpretation of reduced IgG values. Ann Allergy Asthma Immunol, 99(3): 281-283.
6. Collin M, Björck L (2017). Toward Clinical use of the IgG Specific Enzymes IdeS and EndoS against Antibody-Mediated Diseases. Methods Mol Biol, 1535:339-351.
7. Tini M, Jewel UR, Camenisch G, et al (2002). Generation and application of chicken egg –yolk antibodies. Comp Bio-chem Physiol A Mol Integr Physiol, 131(3):569-74.
8. Schade R, Schniering A, Hlinak A (1992). Polyclonal avian antibodies and extracted from egg yolk as an alternative to the production of antibodies in mammals. A review. ALTEX, 9(2): 43-56.
9. Gürtler M, Fehlhaber K (2004). Growth of Salmonella enteritidis in yolk from eggs laid by immunized hens. Int J Food Micro-biol, 90(1):107-13.
10. Gutiérrez Calzado E, García Garrido RM, Schade R (2001). Human haemoclassification by use of specific yolk antibodies obtained after immuniza-tion of chickens against human blood group antigens. Altern Lab Anim, 29(6):717-726.
11. Mc Cannel A, Nakai S(1990). Separation of egg yolk immunoglobulins into sub-populations using DEAE-ion exchange chromatography. Can Inst Food Sci Technol J, 23:46-42.
12. Leslie GA, Clem LW (1969). Phylogen of immunoglobulin structure and function. Immunoglobulins of the chicken. J Exp Med, 130(6):1337-52.
13. Dillon M, Yin Y, Zhou J, et al (2017). Effi-cient production of bispecific IgG of dif-ferent isotypes and species of origin in single mammalian cells. MAbs, 9(2):213-230.
14. Zambrowicz A, Zabłocka Ł, Bobak J, et al (2017). A simple and rapid method of isolation of active polypeptide complex, yolkin, from chicken egg yolk. Food Chem, 230:705-711.
15. Sun S, Mo W, Ji Y, et al (2001). Preparation and mass spectrometric study of egg yolk antibody (IgY) against Rabies virus. Rapid Commun Mass Spectrom, 15(9):708-12.
16. Lee N(2000). Anti-Bacterial Activities of Chicken Egg YolkAntibody (IgY) against Enteric Pathogens. MSc Thesis.University of Alberta, College of Agriculture, Food & Nutritional Science (Canada). 2000.
17. Bižanov G, Normantiene T, Jonauskiene I (2006). Development of Antibodies to Sendi Virus in chickens and their Isola-tion from Yolk. Biologua, 2: 68-71.
18. Arimitsu H, Sasaki K, Kohda T, Shimizu T, Tsuji T (2014). Evaluation of Shiga toxin 2e-specific chicken egg yolk immuno-globulin: Production and neutralization activity. Microbiol Immunol 58(11):643-8.
19. Sheng L, He Zh, Chen J, et al (2017). The impact of N-glycosylation on confor-mation and stability of immunoglobulin Y from egg yolk. Int J Biol Macromol, 96:129-136.
20. Hansen P, Scoble JA, hanson B, et al (1998). Isolation and purification of immuno-globulins from chicken eggs using thi-ophilic interaction chromatography. J Im-munol Methods, 215(1-2):1-7.
21. Walczak M, Grzywa R, Łupicka-Słowik A, et al (2015). Method for generation of pep-tide-specific IgY antibodies directed to Staphylococcus aureus extracellular fi-brinogen binding protein epitope. Biopol-ymers, 104(5):552-9.
22. Mahdavi AH, Rahmani HR, Nili N et al (2010). Effects of dietary Egg Yolk anti-body (IgY) Powder on growth perfor-mance, Intestinal Escherichia Coli coloni-zation, and Immunocompetence of Chal-lenged Broilerchicks. Poult Sci, 89(3):484-94.
23. Pauly D, Chacana PA, Calzado EG et al (2011). IgY technology:extraction of chicken antibodies from egg yolk by polyethyleneglycol (PEG) precipitation. J Vis Exp, (51): 3084.
24. Polson A, von Wechmar MB, van Re-genmortel MH (1980). Isolation of viral IgY antibodies from yolks of immunized hens. Immunol Commun, 9(5):475-93.
25. Ouchterlony O (1949). Antigen-antibody re-actions in gels. Acta Pathol Microbiol Scand, 26(4):507-15.
26. Liu J, He Q, Wang W et al (2017). Prepara-tion and neutralization efficacy of IgY an-tibodies raised against Deinagkistrodon acutus venom. J Venom Anim Toxins Incl Trop Dis, 23:22.
27. Laemmli UK (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, 227(5259):680-5.
28. Tumurjav B, Terkawi MA, Zhang H, et al (2015). Optimization and validation of an ELISA using recombinant Toxoplasma gondii matrix antigen 1 for serodiagnosis of the infection. J Agric Sci, 15(2):56-60.
29. Wilson MB, Nakane PK (1978). Recent de-velopments in the periodate method of conjugating horseradish peroxidase (HRPO) to antibodies, p. 215-224. In W. Knapp, G. Wick, and K. Holubar (ed.), Immunofluorescence and related staining techniques. Elsevier, Amsterdam, The Netherlands.
30. Junior AF, Santiago FM, Silva MV, et al (2012). Production, characterization and applications for Toxoplasma gondii-specific polyclonal chicken egg yolk im-munoglobulins. PLoS One, 7(7):e40391.
31. He J, Hu J, Thirumalai D et al (2016). Devel-opment of indirect competitive ELISA using egg yolk-derived immunoglobulin (IgY) for the detection of Gentamicin residues. J Environ Sci Health B, 51(1):8-13.
32. Ren H, Yang W, Thirumalai D, et al (2016). A comparative evaluation of six principal IgY antibody extraction methods. Alter-natives to laboratory animals: Altern Lab Anim, 44(1):11-20.
33. Priyanka B, Abhijith K, Rastogi NK, et al (2014). Integrated approach for the ex-traction and purification of IgY from chicken egg yolk. Sep Sci Technol, 49(4):562-568.
34. Meenatchisundaram S, Michael A (2010). Comparison of four different purification methods for isolation of anti Echis cari-natus antivenom antibodies from im-munized chicken egg yolk. Iran J Biotechnol, 8(1):50-55.
35. Verdoliva A, Basile G, Fassina G (2000). Af-finity purification of immunoglobulins from chicken egg yolk using a new syn-thetic ligand. J Chromatogr B Biomed Sci Appl, 749(2):233-42.
36. Jiang X, Diraviyam T, Zhang X (2016). Af-finity purification of egg yolk immuno-globulins (IgY) using a human myco-plasma protein. J Chromatogr B Analyt Technol Biomed Life Sci, 1012-1013:37-41.
37. Akita EM, Nakai S (1992). Immunoglobulins from egg yolk: Isolation and purification. J Food Sci, 57(3):629-634.
38. Cook CL, Pao W, Firea JR, et al (2001). Simple purification methods for an α - Galactose-specific antibody from chicken eggs. J Biosci Bioeng, 91(3): 305-310.
39. Deignan J, Kelly A, Gert HJ (2000). Com-parative analysis of methods of purifica-tion of egg yolk immunoglobulin. Food Agric Immunol, 12(1):77-85.
40. Polson A (1990). Isolation of IgY from the yolks of eggs by a chloroform polyeth-ylene glycol procedure. Immunol Invest, 19(3):253-8.
41. Hasan Khan K, Himeno A, Kosugi S, et al (2017). IgY-binding peptide screened from a random peptide library as a ligand for IgY purification. J Pept Sci, 23(10): 790-797.
42. Sudjarwo S, Eraiko K, Sudjarwo GW et al (2017). The potency of chicken egg yolk immunoglobulin (IgY) specific as immu-notherapy to Mycobacterium tuberculosis infection. J Adv Pharm Technol Res, 8(3):91-96.
43. Grover RK, Zhu X, Nieusma T, et al (2014). A structurally distinct human mycoplasma protein that generically blocks antigen-antibody union. Science, 343(6171):656-661.
44. Mulvey GL, Dingle TC, Fang L, et al (2011). Therapeutic potential of egg yolk anti-bodies for treating Clostridium difficile infection. J Med Microbiol, 60(8):1181-7.
45. Witkowski PT, Bourquain DR, Hohn O, et al (2009). Gene gun-supported DNA immunisation of chicken for straightfor-ward production of poxvirus-specifiv IgY antibodies. J Immunol Methods, 341(1-2):146-53.
|Issue||Vol 48 No 7 (2019)|
|Egg yolk HRP Human serum IgY Polyclonal antibody|
|Rights and permissions|
|This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.|