Department of Influenza and other Respiratory Viruses, Pasteur Institute of Iran, Tehran, Iran
Abstract: (3036 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.
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