陶瓷羟基磷灰石一步色谱法纯化登革病毒颗粒

疫苗(英文) Pub Date : 2012-08-03 DOI:10.4236/WJV.2012.23020

Y. Kurosawa, M. Saito, Shintaro Kobayashi, T. Okuyama

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引用次数: 14

摘要

登革病毒2型ThNH7/93经陶瓷羟基磷灰石层析纯化后仍保持感染活性。将登革病毒2型培养液装在陶瓷羟基磷灰石柱上，用线性梯度的磷酸钠缓冲液洗脱。首先洗脱来自宿主细胞的培养液和蛋白质污染物，然后洗脱dsDNA，然后洗脱2型登革热病毒。血凝(HA)法检测登革病毒2型的回收率为64±14% (n = 11)，焦点法检测登革病毒2型的回收率为60% (n = 2)。该方案重复性高，简单，快速，似乎适用于其他病毒种类，如流感病毒，乙型脑炎病毒和腺病毒。

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Purification of Dengue Virus Particles by One-Step Ceramic Hydroxyapatite Chromatography

Dengue virus type 2 ThNH7/93 retained infectious activity after purification by ceramic hydroxyapatite chromatogra-phy. Dengue virus type 2 culture fluid was loaded onto the ceramic hydroxyapatite column and eluted with a linear gradient of sodium phosphate buffer. Culture fluid and protein contaminants derived from host cells were eluted initially, followed by elutions of dsDNA, and then dengue virus type 2. The recoveries of dengue virus type 2 were 64 ± 14% (n = 11) in the hemagglutination (HA) test and 60% (n = 2) determined by focus assay for viral infectivity. This protocol was highly reproducible, simple, rapid, and appears applicable to other virus species such as influenza virus, Japanese encephalitis virus and adenovirus.

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疫苗(英文)

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