Characterization of Anti-E.Coli Antibody Exteracted From Immunized Hen Eggs By Polyethylene Glycol (PEG) Precipitation
-
Mohammad Mehdi Soltan Dallal
,
Monireh Zeinolabedini Zamani
,
Laya Kafami Khorasani
,
Mohammad Hassan Houshdar Tehrani
,
Mohammad Kazem Sharifi Yazdi
,
Saeid Vahedi
,
Farnaz Soltani Jonaghani
,
Fatemehsadat Mirsane
Abstract
Background: The aim of this study was to provide a simple and efficient way of producing IgY antibodies against Escherichia coli and extract that by polyethylene glycol (PEG) precipitation method.
Methods: An ti- Escherichia coli antibody was produced in hens by using formalin-killed E.coli and confirmed by ELISA method, applying serum`antibody. The specific IgY was extracted from egg yolk by Polyethylene Glycol (PEG) Precipitation and analysed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).
Results: The result of ELISA shows that the IgY titre increased from initial immunization and the high titre (≥0.071) persisted after the first immunization. Heavy chain of IgY with ~68 KDa was observed in the Gel electrophoresis pattern.
Conclusion: The results of the ELISA indicates the specificity of the immunoglobulin Y to the target antigen and the result of SDS-PAGE represented the appropriate extraction method. More research must be done on the ability of these antibodies to inhibit the growth of E.coli.
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14. Hirai K, Arimitsu H, Umeda K, et al. Passive oral immunization by egg yolk immunoglobulin (IgY) to Vibrio cholerae effectively prevents cholera. Acta Med Okayama 2010; 64:163-70.
15. Soltan Dallal MM, Kalantar E, Kafami Khorasani L, et al. Identification and extraction of chicken egg yolk immunoglobulin from egg by polyethylene glycol (PEG) precipitation. J Med Bacteriol 2015; 4(5, 6):13-8.
16. Clifton-Hadley FA, Breslin M, Venables LM, et al. A laboratory study of an inactivated bivalent iron restricted Salmonella enterica serovars enteritidis and typhimurium dual vaccine against typhimurium challenge in chickens. Vet Microbiol 2002; 89:167-79.
17. Li C, Ren H, Schade R, et al. A novel and efficient immunoglobulin extraction method using poloxamer-polyethylene glycol. Prep Biochem Biotechnol 2017; 47(7):739-43.
18. Ren H, Yang W, Thirumalai D, et al. A comparative evaluation of six principal IgY antibody extraction methods. Altern Lab Anim 2016; 44(1):11-20.
2. Arathy DS, Vanpee G, Belot G, et al. Antimicrobial drug resistance in Escherichia coli isolated from commercial chicken eggs in Grenada, West Indies. West Indian Med J 2011; 60(1):53-6.
3. George J, Weiner, MD. Building better monoclonal antibody-based therapeutics. Nat Rev Cancer 2015; 15(6):361-70.
4. Zhen YH, Jin LJ, Guo J, et al. Characterization of specific egg yolk immunoglobulin (IgY) against mastitis-causing Staphylococcus aureus. J Appl Microbiol 2008; 105(5):1529-35.
5. Guimarães M, Amara LG, Rangel LBA, et al. Growth inhibition of Staphylococcus aureus by chicken egg yolk antibodies. Arch Immunol Ther Exp 2009; 57:377-82.
6. Meenatchisundaram S, Michael A, Subbraj T, et al. Isolation, purification and neutralizing potential of chicken egg yolk immunoglobulin (IgY) against mastitis causing Escherichia coli in dairy cows in Coimbatore District. Inter J Drug Develop Res 2011; 3:147-53.
7. Liou JF, Shiau JW, Tai C et al. Production of Egg yolk immunoglobulin against Escherichia coli from white leghorn and lohmann chickens. J Animal Vet Advances 2011; 10(18):2349-56.
8. Bozkir A, Hayta G. Preparation and evaluation of multiple emulsions water-in-oil-in-water (w/o/w) as delivery system for influenza virus antigens. J Drug Target 2004; 12(3):157-64.
9. Chakravarthi PV, Shanmugasundaram K, Malmarugan S, et al. In vitro assessment of bacteriostatic potency of egg yolk immunoglobulin against Salmonella typhimurium. J Vet Pub Hlth 2013; 11(1): 39-42.
10. Pauly D, Chacana PA, Calzado EG, et al. IgY Technology: extraction of chicken antibodies from egg yolk by polyethylene glycol (PEG) precipitation. J Vis Exp 2011; 1(51):3084.
11. Laemmeli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227:680-5.
12. Pauly D, Dorner M, Zhang X, et al. Monitoring of laying capacity, immunoglobulin Y concentration, and antibody titer development in chickens immunized with ricin and botulinum toxins over a two-year period. Poult Sci 2009; 88: 281-90.
13. Meulenaer DB, Baert K, Lanckriet H, et al. Development of an enzyme-linked immunosorbent assay for bisphenol A using chicken immunoglobulins. J Agric Food Chem 2002; 50:5273-82.
14. Hirai K, Arimitsu H, Umeda K, et al. Passive oral immunization by egg yolk immunoglobulin (IgY) to Vibrio cholerae effectively prevents cholera. Acta Med Okayama 2010; 64:163-70.
15. Soltan Dallal MM, Kalantar E, Kafami Khorasani L, et al. Identification and extraction of chicken egg yolk immunoglobulin from egg by polyethylene glycol (PEG) precipitation. J Med Bacteriol 2015; 4(5, 6):13-8.
16. Clifton-Hadley FA, Breslin M, Venables LM, et al. A laboratory study of an inactivated bivalent iron restricted Salmonella enterica serovars enteritidis and typhimurium dual vaccine against typhimurium challenge in chickens. Vet Microbiol 2002; 89:167-79.
17. Li C, Ren H, Schade R, et al. A novel and efficient immunoglobulin extraction method using poloxamer-polyethylene glycol. Prep Biochem Biotechnol 2017; 47(7):739-43.
18. Ren H, Yang W, Thirumalai D, et al. A comparative evaluation of six principal IgY antibody extraction methods. Altern Lab Anim 2016; 44(1):11-20.
| Files | ||
| Issue | Vol 8 No 5-6 (2019) | |
| Section | Original Articles | |
| Keywords | ||
| Enzyme-Linked Immunosorbent Assay Escherichia coli Polyethylene Glycol Serum | ||
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How to Cite
1.
Soltan Dallal MM, Zeinolabedini Zamani M, Kafami Khorasani L, Houshdar Tehrani MH, Sharifi Yazdi MK, Vahedi S, Soltani Jonaghani F, Mirsane F. Characterization of Anti-E.Coli Antibody Exteracted From Immunized Hen Eggs By Polyethylene Glycol (PEG) Precipitation. J Med Bacteriol. 2019;8(5-6):38-43.
