Production of polyclonal antibody against Tehran strain influenza virus (A/H1N1/2009) hemagglutinin conserved domain (HA2): brief report
Background: The influenza virus is one of the most important factors for higher morbidity and mortality in the world. Recently, researchers have been focused on influenza conserved antigenic proteins such as hemagglutinin stalk domain (HA2) for vaccine production and serological studies. The HA2 pla...
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doaj-02dbc0066ec64f0d83568eb80d3ddced2020-11-24T22:23:59ZfasTehran University of Medical SciencesTehran University Medical Journal1683-17641735-73222015-10-01737535539Production of polyclonal antibody against Tehran strain influenza virus (A/H1N1/2009) hemagglutinin conserved domain (HA2): brief reportSomayeh Zamani0Fatemeh Fotouhi Chahouki1Zahra Nourmohammadi2Saeideh Sadeghi Neshat3Vahideh Mazaheri4Ali Torabi5Behrokh Farahmand6 Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran. Department of Virology, Influenza Research Lab, Pasteur Institute of Iran, Tehran, Iran. Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran. Department of Virology, Influenza Research Lab, Pasteur Institute of Iran, Tehran, Iran. Department of Virology, Influenza Research Lab, Pasteur Institute of Iran, Tehran, Iran. Department of Virology, Influenza Research Lab, Pasteur Institute of Iran, Tehran, Iran. Department of Virology, Influenza Research Lab, Pasteur Institute of Iran, Tehran, Iran. Background: The influenza virus is one of the most important factors for higher morbidity and mortality in the world. Recently, researchers have been focused on influenza conserved antigenic proteins such as hemagglutinin stalk domain (HA2) for vaccine production and serological studies. The HA2 plays a major role in the fusion of the virus with host cells membrane. The immunity system enables to produce antibody against HA2. The aim of this study is polyclonal antibody production against influenza HA2. Methods: This study was done in the Influenza Research Lab, Pasteur Institute of Iran, Tehran for one year from September 2013 to October 2014. In the present study, recombinant HA2 protein was produced in prokaryotic system and purified using Nickel affinity chromatography. The purified HA2 was mixed with Freund’s adjuvant (complete and incomplete) and injected into two New Zealand white rabbits by intramuscularly and subcutaneously routes. Immunization was continued for several months with two weeks interval. Before each immunization, blood was drawn by venous puncture from the rabbit ear. Function of rabbit's sera was evaluated using radial immunodiffusion (RID) in both forms, Single RID (SRID) and Double RID (DRID). Finally, antiserum activity against HA2 was evaluated using western blotting as serological assay. Results: Sedimentary line and zone was observed in RID assays (SRID and DRID) represent interaction between HA2 protein and anti- HA2 antibody. As well as, western blotting results was positive for HA2 protein. Therefore, these results showed that polyclonal antibody produced against HA2 protein can identify HA2 protein antigenic sites. Conclusion: These findings show that humoral immune responses have properly been stimulated in rabbits and these antibodies can identify HA2 protein and may be suitable for other serological methods.http://tumj.tums.ac.ir/browse.php?a_code=A-10-25-5377&slc_lang=en&sid=1antibody HA2 protein hemagglutinins immunodiffusion influenza virus polyclonal western blotting |
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DOAJ |
language |
fas |
format |
Article |
sources |
DOAJ |
author |
Somayeh Zamani Fatemeh Fotouhi Chahouki Zahra Nourmohammadi Saeideh Sadeghi Neshat Vahideh Mazaheri Ali Torabi Behrokh Farahmand |
spellingShingle |
Somayeh Zamani Fatemeh Fotouhi Chahouki Zahra Nourmohammadi Saeideh Sadeghi Neshat Vahideh Mazaheri Ali Torabi Behrokh Farahmand Production of polyclonal antibody against Tehran strain influenza virus (A/H1N1/2009) hemagglutinin conserved domain (HA2): brief report Tehran University Medical Journal antibody HA2 protein hemagglutinins immunodiffusion influenza virus polyclonal western blotting |
author_facet |
Somayeh Zamani Fatemeh Fotouhi Chahouki Zahra Nourmohammadi Saeideh Sadeghi Neshat Vahideh Mazaheri Ali Torabi Behrokh Farahmand |
author_sort |
Somayeh Zamani |
title |
Production of polyclonal antibody against Tehran strain influenza virus (A/H1N1/2009) hemagglutinin conserved domain (HA2): brief report |
title_short |
Production of polyclonal antibody against Tehran strain influenza virus (A/H1N1/2009) hemagglutinin conserved domain (HA2): brief report |
title_full |
Production of polyclonal antibody against Tehran strain influenza virus (A/H1N1/2009) hemagglutinin conserved domain (HA2): brief report |
title_fullStr |
Production of polyclonal antibody against Tehran strain influenza virus (A/H1N1/2009) hemagglutinin conserved domain (HA2): brief report |
title_full_unstemmed |
Production of polyclonal antibody against Tehran strain influenza virus (A/H1N1/2009) hemagglutinin conserved domain (HA2): brief report |
title_sort |
production of polyclonal antibody against tehran strain influenza virus (a/h1n1/2009) hemagglutinin conserved domain (ha2): brief report |
publisher |
Tehran University of Medical Sciences |
series |
Tehran University Medical Journal |
issn |
1683-1764 1735-7322 |
publishDate |
2015-10-01 |
description |
Background: The influenza virus is one of the most important factors for higher morbidity and mortality in the world. Recently, researchers have been focused on influenza conserved antigenic proteins such as hemagglutinin stalk domain (HA2) for vaccine production and serological studies. The HA2 plays a major role in the fusion of the virus with host cells membrane. The immunity system enables to produce antibody against HA2. The aim of this study is polyclonal antibody production against influenza HA2. Methods: This study was done in the Influenza Research Lab, Pasteur Institute of Iran, Tehran for one year from September 2013 to October 2014. In the present study, recombinant HA2 protein was produced in prokaryotic system and purified using Nickel affinity chromatography. The purified HA2 was mixed with Freund’s adjuvant (complete and incomplete) and injected into two New Zealand white rabbits by intramuscularly and subcutaneously routes. Immunization was continued for several months with two weeks interval. Before each immunization, blood was drawn by venous puncture from the rabbit ear. Function of rabbit's sera was evaluated using radial immunodiffusion (RID) in both forms, Single RID (SRID) and Double RID (DRID). Finally, antiserum activity against HA2 was evaluated using western blotting as serological assay. Results: Sedimentary line and zone was observed in RID assays (SRID and DRID) represent interaction between HA2 protein and anti- HA2 antibody. As well as, western blotting results was positive for HA2 protein. Therefore, these results showed that polyclonal antibody produced against HA2 protein can identify HA2 protein antigenic sites. Conclusion: These findings show that humoral immune responses have properly been stimulated in rabbits and these antibodies can identify HA2 protein and may be suitable for other serological methods. |
topic |
antibody HA2 protein hemagglutinins immunodiffusion influenza virus polyclonal western blotting |
url |
http://tumj.tums.ac.ir/browse.php?a_code=A-10-25-5377&slc_lang=en&sid=1 |
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