RT - Journal Article T1 - Production and Evaluation of Polyclonal Antibody Against Influenza A Virus Matrix 2 Conserved Protein for Research and Diagnosis Purposes JF - virusj YR - 2017 JO - virusj VO - 11 IS - 4 UR - http://journal.isv.org.ir/article-1-335-en.html SP - 31 EP - 38 K1 - ELISA K1 - Influenza Virus K1 - M2 protein K1 - polyclonal antibody K1 - RID. AB - 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. LA eng UL http://journal.isv.org.ir/article-1-335-en.html M3 ER -