Related Experiment Video
Updated: Apr 11, 2026
Optimization of Antigen Preparation for Hemagglutination Inhibition Assay of Newcastle Disease Virus Serology
Published on: January 24, 2025
Development of hen antihepatitis B antigen IgY-based conjugate for ELISA assay
Najat Muayed Nafea1, Majeed Arsheed Sabbah2, Raghad Al-Suhail1
1Department of Biology, College of Science, Baghdad University, Isfahan, Iran.
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.
