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Development of hen antihepatitis B antigen IgY-based conjugate for ELISA assay.

Posted by on in 2015
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Nafea NM1Sabbah MA2Al-Suhail R1Mahdavi AH3Asgary S4. 2015. Adv Biomed Res. 4:100. doi: 10.4103/2277-9175.156678. eCollection 2015.

1Department of Biology, College of Science, Baghdad University, Isfahan, Iran.
2Biotechnology Research Center, Alnahrain University, Iraq.
3Department of Animal Sciences, College of Agriculture, Isfahan University of Technology, Isfahan, Iran.
4Cardiovascular Research Center, Isfahan Cardiovascular Research Institute, Physiology Research Center, Isfahan University of Medical Sciences, Isfahan, Iran.



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 polyclonalIgY against hepatitis virus surface antigen (HBsAg) were carried out.


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.


A highly purified of 180 KDa with specific activity of 200 mIU/ml was obtained by our purification protocol. One milligram of the purified IgYwas 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.


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.


ELISA; IgY; hens; hepatitis B; immunization

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