| ORIGINAL ARTICLE |
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| Year : 2015 | Volume
: 4
| Issue : 1 | Page : 100 |
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Development of hen antihepatitis B antigen IgY-based conjugate for ELISA assay
Najat Muayed Nafea1, Majeed Arsheed Sabbah2, Raghad AL-Suhail1, Amir Hossein Mahdavi3, Sedigheh Asgary4
1 Department of Biology, College of Science, Baghdad University, Iran
2 Biotechnology Research Center, Alnahrain University, Iran
3 Department of Animal Sciences, College of Agriculture, Isfahan University of Technology, Isfahan, Iran
4 Cardiovascular Research Center, Isfahan Cardiovascular Research Institute, Physiology Research Center, Isfahan University of Medical Sciences, Isfahan, Iran
Correspondence Address:
Amir Hossein Mahdavi
Department of Animal Sciences, College of Agriculture, Isfahan University of Technology, Isfahan
Iran
 Source of Support: None, Conflict of Interest: None  | Check |
DOI: 10.4103/2277-9175.156678
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Background: Chicken antibodies have many advantages to the mammalian antibodies and have several important differences against mammalian IgG with regard to their specificity and large-scale production. In this study, the production, purification, and HRP conjugation of polyclonal IgY against hepatitis virus surface antigen (HBsAg) were carried out.
Materials and Methods: Single Comb White Leghorn hens were immunized intramuscularly with hepatitis B vaccine in combination with Freund's adjuvants. Blood and eggs were collected before and during ten weeks after the first immunization.
Results: A highly purified of 180 KDa with specific activity of 200 mIU/ml was obtained by our purification protocol. One milligram of the purified IgY was labeled with horseradish peroxidase (HRP). Sandwich ELISA was used to determine the optimum titer of anti-HbsAg IgY-conjugate which was found to be 1:20.
Conclusions: This study showed that laying hens can be used as an alternative source for production of polyclonal antibodies against HBsAg and anti-HBs IgY could be labeled with HRP enzyme and could subsequently be used successfully as secondary antibody in ELISA for detection of HBsAg in the patients sera. |
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