通过亲和层析和非离子密度梯度离心法纯化具有生物活性的爱泼斯坦-巴尔病毒。

Purification of biologically active Epstein-Barr virus by affinity chromatography and non-ionic density gradient centrifugation.

作者信息

Fowler E, Raab-Traub N, Hester S

出版信息

J Virol Methods. 1985 May;11(1):59-74. doi: 10.1016/0166-0934(85)90125-9.

Abstract

Epstein-Barr virus was purified by affinity chromatography on ricin agglutinin Sepharose followed by non-ionic density gradient centrifugation on Nycodenz. The purified virus was highly active in three biological assays: stimulation of immunoglobulin synthesis by B lymphocytes, transformation of B lymphocytes, and superinfection of Raji cells, as well as in an indirect immunofluorescent binding assay. Electron microscopy revealed intact viral particles free of contaminating membranous structures. CsCl density gradient analysis of nick-translated DNA showed material only at the expected viral density with no detectable material at the density of cellular DNA. SDS gel electrophoresis of radioiodinated virus revealed the characteristic viral polypeptides. This method produces highly purified, biologically active virions with an overall recovery of about 30%.

摘要

通过在蓖麻凝集素琼脂糖上进行亲和层析，随后在 Nycodenz 上进行非离子密度梯度离心来纯化爱泼斯坦 - 巴尔病毒。纯化后的病毒在三种生物学检测中具有高活性：刺激 B 淋巴细胞合成免疫球蛋白、B 淋巴细胞转化以及 Raji 细胞的超感染，同时在间接免疫荧光结合检测中也表现出高活性。电子显微镜显示完整的病毒颗粒，无膜结构污染物。对切口平移 DNA 进行 CsCl 密度梯度分析表明，仅在预期的病毒密度处有物质，在细胞 DNA 密度处未检测到物质。对放射性碘化病毒进行 SDS 凝胶电泳显示出特征性的病毒多肽。该方法可产生高度纯化、具有生物活性的病毒粒子，总体回收率约为 30%。