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Iran J Virol 2017, 11(4): 31-38 Back to browse issues page
Production and Evaluation of Polyclonal Antibody Against Influenza A Virus Matrix 2 Conserved Protein for Research and Diagnosis Purposes
S Zamani , F Fotouhi Chahouki , P Mehrbod , S Sadeghi Neshat , B Farahmand
Department of Influenza and other Respiratory Viruses, Pasteur Institute of Iran, Tehran, Iran
Abstract:   (1949 Views)
Background and Aims: The aim of this study was to produce and purify the Polyclonal antibody (pAb) against Matrix protein 2 (M2) with reasonable efficiency. Matrix protein 2 is one of the most conserved proteins of the influenza A virus which acts as ion channel. Polyclonal antibodyproduced against Matrix protein 2 is used in vaccine research, passive immunization and qualitative/quantitative analysis methods.
Materials and Methods: Recombinant M2 protein was produced in E.coli. Purified protein with Freund’s adjuvants (Complete and Incomplete) was injected into two New-Zealand white male rabbits. The polyclonal antisera of rabbits were evaluated by RID, immunoblotting and ELISA. The IgG was purified using DAEA-cellulose column chromatography. Finally, the quality and properties of purified IgG were evaluated using SDS-PAGE and ELISA.
Results: The RID and immunoblotting results showed that the produced anti-M2 antibody was able to recognize M2 recombinant protein epitopes. The ELISA results confirmed anti-M2 pAb reached reasonable titers after three injections. IgG against M2 was purified with suitable concentration. The Purified polyclonal IgG-M2 was evaluated using ELISA and the results showed IgG-M2 reacted with the antigen up to 1:32000.
Conclusions: The data showed that recombinant M2 protein was able to stimulate immune response to produce antibody at satisfactory level.
Keywords: ELISA, Influenza Virus, M2 protein, polyclonal antibody, RID.
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Type of Study: Original article | Subject: Special
Received: 2018/11/16 | Accepted: 2019/01/26 | Published: 2019/01/26
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Zamani S, Fotouhi Chahouki F, Mehrbod P, Sadeghi Neshat S, Farahmand B. Production and Evaluation of Polyclonal Antibody Against Influenza A Virus Matrix 2 Conserved Protein for Research and Diagnosis Purposes. Iran J Virol. 2017; 11 (4) :31-38
URL: http://journal.isv.org.ir/article-1-335-en.html

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