Purification of neuraminidase from Influenza virus subtype H5N1

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dc.creatorTarigan, Simson
dc.creatorIndryani, Risa
dc.creator., Darminto
dc.creatorIgnjatovic, Jagoda
dc.date2013-05-07
dc.date.accessioned2018-06-04T06:46:53Z
dc.date.available2018-06-04T06:46:53Z
dc.date.issued2013-05-07
dc.descriptionInfluenza-virus neuraminidase plays vital role in the survival of the organisms. Vaccination of animals with this glycoprotein confers immune responses so that enable it to protect the animals from incoming infection. Supplementation of conventional vaccines with this glycoprotein increases the protection and longevity of the vaccine. Purified neuraminidase can also be used to develop serological tests for differentiation of serologically positive animals due to infection or to vaccination. In this study purification of neuraminidase from influenza virus subtype H5N1 was described. Triton x-100 and Octyl β-D-glucopyranoside were used to extract and diluted the glycoprotein membrane. The enzymatic activity of the neuraminidase was assayed using a fluorochrome substrate, 4-methylumbelliferyl-a-D-N-acetyl neuraminic acid, which was found to be simple, sensitive and suitable for the purification purpose. The neuraminidase was absorbed selectively on an oxamic-acid agarose column. The purity of neuraminidase eluted from this affinity column was high. A higher purity of the neuraminidase was obtained by further separation with gel filtration on Superdex-200. The purified neuraminidase was enzymatically active and did not contain any detectable haemagglutinin, either by haemagglutination assay or by monospecific antibodies raised against H5N1 hemagglutinin.  The purified neuraminidase was recognized strongly by antibodies raised against an internal but only weakly by that against C-terminal regions of the neuraminidase protein of H5N1-influenza virus. The purified neuraminidase was in tetrameric forms but dissociated into monomeric form on reducing condition, or mostly dimeric form on non-reducing SDS-PAGE. Key Words: Neuraminidase, Influenza, H5N1, Methylumbelliferyl, Oxamic-aciden-US
dc.formatapplication/pdf
dc.identifierhttp://medpub.litbang.pertanian.go.id/index.php/jitv/article/view/366
dc.identifier10.14334/jitv.v14i1.366
dc.identifier.urihttps://repository.pertanian.go.id/handle/123456789/2656
dc.languageeng
dc.publisherIndonesian Animal Sciences Societyen-US
dc.relationhttp://medpub.litbang.pertanian.go.id/index.php/jitv/article/view/366/375
dc.rightsCopyright (c) 1970 Indonesian Journal of Animal and Veterinary Sciencesen-US
dc.rightshttp://creativecommons.org/licenses/by/4.0en-US
dc.source2252-696X
dc.source0853-7380
dc.sourceIndonesian Journal of Animal and Veterinary Sciences; Vol 14, No 1 (2009): MARCH 2009; p.75-82en-US
dc.titlePurification of neuraminidase from Influenza virus subtype H5N1en-US
dc.typeinfo:eu-repo/semantics/article
dc.typeinfo:eu-repo/semantics/publishedVersion
dc.typePeer-reviewed Articleen-US
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