2013•Iranian Journal of VirologyOpen access

Characterization of Nucleoprotein Extracted from Human Influenza A Virus Cultured in Two Different Cell Lines

Yasin Panahi, Behrokh Farahmand, Rasoul Soleimani-Stiar, Reza Saghiri, SHima Fattahi, Mansoureh Tabatabaeian, Masoumeh Tavassoti-Kheiri

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Abstract

Background and Aims: Influenza virus nucleoprotein (NP) has the capacity to be used as subunit vaccine, but little is known about the impact of different cultures on its structure.In the present study we aimed to evaluate and compare the Isoelectric focusing (IEF) property of extracted viral nucleoproteins derived from Madin Darby canine kidney (MDCK) cell line and embryonated chicken eggs (ECE). Materials and Methods:Influenza virus strain A/NewCaledonia/20/99/H1N1 was propagated and grown in allantoic sac of 10-11 day-old embryonated chicken eggs, and mammalian cell culture (MDCK) in parallel.Ribonucleoprotein extraction was conducted from two separate cultures and evaluated using isoelectric focusing gel strips. Results:The results showed higher isoelectric pH in extracted nucleoproteins from MDCK as compared to embryonated chicken eggs.Conclusion: It is possible that some amino acids have been replaced.Suggesting that the changing net charge of protein may be affect the conserved regions of the protein.Therefore, this could impact the new generation of vaccines construction based on conserved proteins.

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Background and Aims: Influenza virus nucleoprotein (NP) has the capacity to be used as subunit vaccine, but little is known about the impact of different cultures on its structure.In the present study we aimed to evaluate and compare the Isoelectric focusing (IEF) property of extracted viral nucleoproteins derived from Madin Darby canine kidney (MDCK) cell line and embryonated chicken eggs (ECE). Materials and Methods:Influenza virus strain A/NewCaledonia/20/99/H1N1 was propagated and grown in allantoic sac of 10-11 day-old embryonated chicken eggs, and mammalian cell culture (MDCK) in parallel.Ribonucleoprotein extraction was conducted from two separate cultures and evaluated using isoelectric focusing gel strips. Results:The results showed higher isoelectric pH in extracted nucleoproteins from MDCK as compared to embryonated chicken eggs.Conclusion: It is possible that some amino acids have been replaced.Suggesting that the changing net charge of protein may be affect the conserved regions of the protein.Therefore, this could impact the new generation of vaccines construction based on conserved proteins.

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Available abstract

Background and Aims: Influenza virus nucleoprotein (NP) has the capacity to be used as subunit vaccine, but little is known about the impact of different cultures on its structure.In the present study we aimed to evaluate and compare the Isoelectric focusing (IEF) property of extracted viral nucleoproteins derived from Madin Darby canine kidney (MDCK) cell line and embryonated chicken eggs (ECE). Materials and Methods:Influenza virus strain A/NewCaledonia/20/99/H1N1 was propagated and grown in allantoic sac of 10-11 day-old embryonated chicken eggs, and mammalian cell culture (MDCK) in parallel.Ribonucleoprotein extraction was conducted from two separate cultures and evaluated using isoelectric focusing gel strips. Results:The results showed higher isoelectric pH in extracted nucleoproteins from MDCK as compared to embryonated chicken eggs.Conclusion: It is possible that some amino acids have been replaced.Suggesting that the changing net charge of protein may be affect the conserved regions of the protein.Therefore, this could impact the new generation of vaccines construction based on conserved proteins.

Key concepts: Nucleoprotein, Virology, Virus, Biology, Cell culture, Influenza A virus, Cell, Orthomyxoviridae

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