Advanced Biomedical Research 2277-9175 2015 May 2015 05 01 Development of hen antihepatitis B antigen IgY-based conjugate for ELISA assay 1 6 30323 EN Najat Muayed Nafea Department of Biology, College of Science, Baghdad University, Iran Majeed Arsheed Sabbah Biotechnology Research Center, Alnahrain University, Iran Raghad AL-Suhail Department of Biology, College of Science, Baghdad University, Iran Amir Hossein Mahdavi Department of Animal Sciences, College of Agriculture, Isfahan University of Technology, Isfahan, Iran Sedigheh Asgary Cardiovascular Research Center, Isfahan Cardiovascular Research Institute, Physiology Research Center, Isfahan University of Medical Sciences, Isfahan, Iran Journal Article 2023 03 29 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.

https://advbiores.mui.ac.ir/article_30323_99a3812141d4296fa601af58724d8a7f.pdf