فهرست مطالب f. fotouhi chahouki
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Background and AimsThe 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 MethodsRecombinant 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.ResultsThe 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.ConclusionsThe data showed that recombinant M2 protein was able to stimulate immune response to produce antibody at satisfactory levelKeywords: ELISA, Influenza Virus, M2 protein, polyclonal antibody, RID.}
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