Gene Medicine Research Group, NDCLS, Radcliffe Department of Medicine, John Radcliffe Hospital, Oxford University, Oxford, UK.
United Kingdom Cystic Fibrosis Gene Therapy Consortium, Oxford, Edinburgh, London, UK.
Sci Rep. 2017 Aug 31;7(1):10198. doi: 10.1038/s41598-017-07893-3.
The development of lentiviral-based therapeutics is challenged by the high cost of current Good Manufacturing Practices (cGMP) production. Lentiviruses are enveloped viruses that capture a portion of the host cell membrane during budding, which then constitutes part of the virus particle. This process might lead to lipid and protein depletion in the cell membrane and affect cell viability. Furthermore, growth in suspension also causes stresses that can affect virus production yields. To assess the impact of these issues, selected supplements (Cholesterol Lipid Concentrate, Chemically Defined Lipid Concentrate, Lipid Mixture 1, Gelatin Peptone N3, N-Acetyl-L-Cysteine and Pluronic F-68) were assayed in order to improve production yields in a transient transfection production of a Sendai virus F/HN-pseudotyped HIV-1-based third generation lentiviral vector in FreeStyle 293 (serum-free media) in suspension. None of the supplements tested had a significant positive impact on lentiviral vector yields, but small non-significant improvements could be combined to increase vector production in a cell line where other conditions have been optimised.
基于慢病毒的治疗方法的发展受到当前良好生产规范 (cGMP) 生产的高成本的挑战。慢病毒是包膜病毒,在出芽过程中捕获宿主细胞膜的一部分,然后构成病毒颗粒的一部分。这个过程可能导致细胞膜中的脂质和蛋白质耗竭,并影响细胞活力。此外,悬浮培养也会造成影响病毒生产产量的压力。为了评估这些问题的影响,选择了几种补充剂(胆固醇脂质浓缩物、化学定义的脂质浓缩物、脂质混合物 1、明胶胨 N3、N-乙酰-L-半胱氨酸和泊洛沙姆 F-68)进行了检测,以便在悬浮液中的 FreeStyle 293(无血清培养基)中转染瞬时转染生产的一株新城疫病毒 F/HN-假型 HIV-1 第三代慢病毒载体时提高生产产量。测试的补充剂均未对慢病毒载体产量产生显著的积极影响,但可以将小的非显著改善结合起来,以提高其他条件已优化的细胞系中的载体生产。