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Evaluation of Retroviral Production Systems Using Quantitative Analysis

机译:使用定量分析评估逆转录病毒生产系统

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Because of the low titer of retroviral supernatant it is necessary to develop and optimize large-scale retriviral production systems.To quantitatively determine the effect of a given operation condiation(e.g.temperature and serum content) on producer cells' retrovirus-producing apacity,a mathematical model was used to analyze the static retroviral production system described by three processes viral diffusion decay and generation.The analytical solutions of the defined model equations were fitted with experimental data to determine the specific retroviral production rate constant which represents the competence of a retroviral production system.Two different retroviral production systems inducible production of vesicular stomatitis virus G glycopretein (VSV-G) pseudotyped retrovirus from 293GPG/EGFPcells and constant production of ecotropic retrovirus from GP+E86/LNCX cells were empolyed to demonstarate the feasibility of the engineering analysis Our results indicated that the time-variant specific retroviral production rate of 293gpg/egfp cells reached its maximum value of 5.7 x 10~-3 CFU/cm2.h.cell and the constant specific rtroviral production rate fo GP+E86/LNCX cells was 1.49 X10~-2 CFU/cm2.h.cell for the whole period of productionFurthermore the effects of serum concentration and temperature on the ecotropic retriviral production system were examined separately.Our results suggest that producting ecotropic recovirused with 10% fetal bovine serum at 37 deg C is the optiml operating conditions for the long-term production system used here.
机译:由于逆转录病毒上清液的滴度很低,因此有必要开发和优化大规模的逆转录病毒生产系统。要定量确定给定的操作条件(例如温度和血清含量)对生产细胞产生逆转录病毒的能力的影响,该模型用于分析由病毒扩散衰减和产生的三个过程描述的静态逆转录病毒生产系统。对定义的模型方程的解析解与实验数据进行拟合,以确定代表逆转录病毒生产系统能力的特定逆转录病毒生产速率常数。强调了两种不同的逆转录病毒生产系统,可从293GPG / EGFP细胞诱导产生水泡性口炎病毒G糖蛋白(VSV-G)假型逆转录病毒,并从GP + E86 / LNCX细胞不断产生嗜酸性逆转录病毒,以证明工程分析的可行性。表示时变293gpg / egfp细胞的特异逆转录病毒生产速率达到最大值5.7 x 10〜-3 CFU / cm2.h.cell,GP + E86 / LNCX细胞的恒定反转录病毒生产速率为1.49 X10〜-2 CFU /整个生产过程中的cm2.h.cell此外,还分别检查了血清浓度和温度对亲热逆转录病毒生产系统的影响。我们的结果表明,在37°C的条件下以10%胎牛血清生产亲和性再病毒是最佳操作条件对于这里使用的长期生产系统。

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