10-Year stability of clinical-grade serum-free γ-retroviral vector-containing medium
10-Year stability of clinical-grade serum-free γ-retroviral vector-containing medium
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DOI：
10.1038/gt.2010.126
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发表时间：
2011-02-01
期刊：
影响因子：
5.1
通讯作者：
Glimm, H.
中科院分区：
文献类型：
--
作者：
Herbst, F.;Ball, C. R.;Glimm, H.
十多年前，我们开发了一种有效的方案，用于对来自动员的外周血的人类造血干细胞进行无血清逆转录病毒转导。方法升级后，在严格的良好生产规范 (GMP) 条件下生产的无血清逆转录病毒长臂猿白血病病毒 (GALV) 假型 PG13/LN 载体上清液用于德国首次临床基因标记试验。在本研究中，我们分析了这种无血清临床级逆转录病毒上清液生产 10 年后的滴度和转导效率，以评估其长期稳定性。干冰上的长期储存和运输导致CD34(+)细胞的滴度和转导效率水平略有下降，范围为38.4%至49.1%。我们的结论是，无血清培养基中逆转录病毒载体的稳定性允许在多中心临床试验中延长批准的临床级逆转录病毒载体库存的储存和分发到遥远的地点。基因治疗 (2011) 18, 210-212; doi：10.1038/gt.2010.126； 2010 年 11 月 11 日在线发布
More than 10 years ago, we developed an efficient protocol for serum-free retroviral transduction of human hematopoietic stem cells derived from mobilized peripheral blood. After upscaling of the methodology, serum-free retroviral gibbon-ape leukemia virus (GALV) pseudotype PG13/LN vector supernatant produced under strict good manufacturing practice (GMP) conditions was used in the first clinical gene-marking trial in Germany. In this study, we analyzed the titer and transduction efficiency of this serum-free clinical-grade retroviral supernatant 10 years after production to evaluate the long-term stability. Long-term storage and transport on dry ice resulted in modestly decreased titers and levels of transduction efficiency in CD34(+) cells ranging from 38.4 to 49.1%. We conclude that the stability of retroviral vectors in serum-free medium allows extended storage and distribution of approved clinical-grade retroviral vector stocks to distant sites in multicenter clinical trials. Gene Therapy (2011) 18, 210-212; doi:10.1038/gt.2010.126; published online 11 November 2010