PlasmidFactory GmbH & Co. KG , Bielefeld, Germany .
Hum Gene Ther. 2017 Oct;28(10):856-861. doi: 10.1089/hum.2017.159. Epub 2017 Aug 21.
Plasmid DNA is currently gaining increasing importance for clinical research applications in gene therapy and genetic vaccination. For direct gene transfer into humans, good manufacturing practice (GMP)-grade plasmid DNA is mandatory. The same holds true if the drug substance contains a genetically modified cell, for example chimeric antigen receptor (CAR) T cells, where these cells as well as the contained plasmids are used. According to the responsible regulatory agencies, they have to be produced under full GMP. On the other hand, for GMP production of, for example, mRNA or viral vectors (lentiviral vectors, adeno-associated virus vectors, etc.), in many cases, High Quality Grade plasmid DNA is accepted as a starting material. The manufacturing process passes through different production steps. To ensure the right conditions are used for the plasmid, a pilot run must be conducted at the beginning. In this step, a followed upscaling with respect to reproducibility and influences on product quality is performed. Subsequently, a cell bank of the transformed productions strain is established and characterized. This cell bank is used for the cultivation process. After cell harvesting and lysis, several chromatography steps are conducted to receive a pure plasmid product. Depending on the respective required quality grade, the plasmid product is subject to several quality controls. The last step consists of formulation and filling of the product.
质粒 DNA 目前在基因治疗和基因疫苗的临床研究应用中越来越重要。对于直接将基因转入人体，必须使用良好生产规范 (GMP) 级别的质粒 DNA。如果药物成分含有经过基因改造的细胞，例如嵌合抗原受体 (CAR) T 细胞，那么这些细胞以及包含的质粒也将被使用。根据负责的监管机构的要求，这些细胞和质粒都必须在完全 GMP 条件下生产。另一方面，对于例如 mRNA 或病毒载体（慢病毒载体、腺相关病毒载体等）的 GMP 生产，在许多情况下，高纯度等级的质粒 DNA 可作为起始材料接受。生产过程需要经过不同的生产步骤。为了确保质粒处于正确的条件下，必须在开始时进行试验性放大，以确保重现性和对产品质量的影响。随后，建立和表征转化生产菌株的细胞库。该细胞库用于培养过程。细胞收获和裂解后，进行几个色谱步骤以获得纯质粒产品。根据各自所需的质量等级，质粒产品需要进行几次质量控制。最后一步包括产品的配方和灌装。
