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通过单克隆抗体亲和层析从细胞培养物中纯化传染性犬细小病毒。

Purification of infectious canine parvovirus from cell culture by affinity chromatography with monoclonal antibodies.

作者信息

Rimmelzwaan G F, Groen J, Juntti N, Teppema J S, UytdeHaag F G, Osterhaus A D

出版信息

J Virol Methods. 1987 Mar;15(4):313-22. doi: 10.1016/0166-0934(87)90154-6.

Abstract

Immuno affinity chromatography with virus neutralizing monoclonal antibodies, directed to the haemagglutinating protein of canine parvovirus (CPV) was used to purify and concentrate CPV from infected cell culture. The procedure was monitored by testing the respective fractions in an infectivity titration system, in an ELISA, in a haemagglutination assay and by negative contrast electron microscopy to quantify CPV or CPV antigen. The degree of purification was further estimated by testing the fractions for total protein content in a colorimetric method, for bovine serum albumin content in an ELISA and by SDS-PAGE. Over 99% of the contaminating proteins proved to be removed, and 20% or 70-90% of infectious CPV or CPV antigen, respectively, was recovered.

摘要

使用针对犬细小病毒(CPV)血凝蛋白的病毒中和单克隆抗体进行免疫亲和层析,从感染的细胞培养物中纯化并浓缩CPV。通过在感染性滴定系统、酶联免疫吸附测定(ELISA)、血凝试验中检测各个组分,并通过负染电子显微镜对CPV或CPV抗原进行定量,来监测该过程。通过比色法检测组分中的总蛋白含量、ELISA检测牛血清白蛋白含量以及十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳(SDS - PAGE),进一步估计纯化程度。结果表明,超过99%的污染蛋白被去除,分别回收了20%的感染性CPV或70 - 90%的CPV抗原。

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