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在新建立的细胞系中诱导产生并部分纯化大鼠成纤维细胞源性干扰素

Production and partial purification of rat fibroblast-derived interferon induced in a newly established cell line.

作者信息

Davis C T, Rusckowski M, Goldstein D A, Murasko D M

出版信息

J Interferon Res. 1984 Summer;4(3):365-74. doi: 10.1089/jir.1984.4.365.

Abstract

Interferon (IFN) production induced by either Newcastle disease virus (NDV) or Sendai virus was compared in 10 different rat cell lines. Although there was variation in the IFN titers produced, NDV proved to be the best inducer in each cell line with optimum IFN yields occurring with a multiplicity of infection (MOI) of 1.0. A new continuous rat fibroblast cell line (CD) produced high titers of IFN similar to those reported for other high producers; while Ratec cells were shown to be the most sensitive to the antiviral activity of rat IFN. Partial purification and characterization of IFN produced in CD cells was accomplished by column chromatography. Four sorbents with varying modes of action for binding the IFN (Affi-Gel 202, Poly(U)-Sepharose 4B, CM-Sepharose CL-6B, and Phenyl-Sepharose CL-4B) were compared. The Phenyl-Sepharose CL-4B matrix proved to be the most successful for purification of rat IFN; one passage through this column, increased the specific activity more than 100-fold, with a concomitant recovery of 90%-95% biologic activity. The binding characteristics of rat IFN on each of the column matrices, however, demonstrated differences between the physicochemical nature of CD rat IFN and murine and hamster IFNs.

摘要

在10种不同的大鼠细胞系中比较了新城疫病毒(NDV)或仙台病毒诱导产生的干扰素(IFN)。尽管产生的IFN效价存在差异,但在每种细胞系中,NDV被证明是最佳诱导剂,当感染复数(MOI)为1.0时,IFN产量达到最佳。一种新的连续大鼠成纤维细胞系(CD)产生的IFN效价很高,与其他高产细胞系报道的效价相似;而Ratec细胞被证明对大鼠IFN的抗病毒活性最为敏感。通过柱色谱法对CD细胞中产生的IFN进行了部分纯化和特性鉴定。比较了四种具有不同IFN结合作用方式的吸附剂(Affi-Gel 202、聚(U)-琼脂糖凝胶4B、CM-琼脂糖凝胶CL-6B和苯基-琼脂糖凝胶CL-4B)。结果表明,苯基-琼脂糖凝胶CL-4B基质对大鼠IFN的纯化最为成功;通过该柱一次,比活性提高了100多倍,同时生物活性回收率为90%-95%。然而,大鼠IFN在每种柱基质上的结合特性表明,CD大鼠IFN与小鼠和仓鼠IFN的物理化学性质存在差异。

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