development of hen antihepatitis b antigen igy-based conjugate for elisa assay

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ID: 164604
2015
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Abstract
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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nafea2015advanceddevelopment Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors ;Najat Muayed Nafea;Majeed Arsheed Sabbah;Raghad AL-Suhail;Amir Hossein Mahdavi;Sedigheh Asgary
Journal microbiological research
Year 2015
DOI
10.4103/2277-9175.156678
URL
http://www.advbiores.net/article.asp?issn=2277-9175;year=2015;volume=4;issue=1;spage=100;epage=100;aulast=Nafea
Keywords
ELISA hepatitis b

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