...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Journal of chromatography, B. Analytical technologies in the biomedical and life sciences >High-performance liquid chromatographic total particles quantification of retroviral vectors pseudotyped with vesicular stomatitis virus-G glycoprotein
【24h】

High-performance liquid chromatographic total particles quantification of retroviral vectors pseudotyped with vesicular stomatitis virus-G glycoprotein

机译：水泡性口腔炎病毒-G糖蛋白假型逆转录病毒载体的高效液相色谱总颗粒定量

获取原文
获取原文并翻译 | 示例
   

获取外文期刊封面封底 >>

       
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

A novel and rapid method for the total particles quantification of murine leukemia virus derived retroviral vectors pseudotyped with vesicular stomatitis virus-G glycoprotein was developed using high performance liquid chromatography. Virus particles were detected by absorbance at 260 nm and quantified using a calibration curve generated from highly purified and concentrated viral stock characterized by negative stain electron microscopy. The method requires Benzonase(R) digestion and concentration of the supernatant prior to analysis. The virus eluted in 12.55 min at a flow rate of 1 mL/min in 20 mM Tris-Cl, pH 7.4 + 1.1 M NaCl. The limits of detection and quantification of this assay were 4.71 x 10(8) and 1.57 x 10(9) viral particles/mL, respectively. Linearity was between 3.0 x 10(9) and 1.0 x 10(11) viral particles/mL with a correlation coefficient of 0.9923 and a slope of 6 x 10(-6). The assay precision was < 5 % and < 1 0% for intra- and inter-day analysis, respectively. This assay was used for the total particles quantification of a 7-day, large-scale perfusion culture production of a retroviral vector grown in 293 cells expressing the P-galactosidase gene. (C) 2004 Published by Elsevier B.V.
机译：利用高效液相色谱法，开发了一种新型，快速的方法，用于以假性水疱性口炎病毒-G糖蛋白假型的鼠白血病病毒衍生的逆转录病毒载体的总颗粒定量。通过在260nm处的吸光度检测病毒颗粒，并使用由以负染色电子显微镜表征的高度纯化和浓缩的病毒原液产生的校准曲线进行定量。该方法需要Benzonase消化并在分析之前浓缩上清液。病毒在12.55分钟内以1 mL / min的流速在20 mM Tris-Cl（pH 7.4 + 1.1 M NaCl）中洗脱。该测定的检测和定量限分别为4.71 x 10（8）和1.57 x 10（9）病毒颗粒/ mL。线性介于3.0 x 10（9）和1.0 x 10（11）病毒颗粒/ mL之间，相关系数为0.9923，斜率为6 x 10（-6）。日内和日间分析的分析精度分别为<5％和<1 0％。该测定法用于在表达P-半乳糖苷酶基因的293细胞中生长的逆转录病毒载体的7天大规模灌注培养生产的总颗粒定量。 （C）2004由Elsevier B.V.发布

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号