Radhakrishnan Harikrishnan, Javitz Harold S, Bhatnagar Parijat
Biosciences Division, SRI International, Menlo Park, CA, 94025, USA.
Education Division, SRI International, Menlo Park, CA, 94025, USA.
Adv Biosyst. 2020 Jun;4(6):e1900288. doi: 10.1002/adbi.201900288. Epub 2020 May 10.
A process for maximizing the titer of lentivirus particles, deemed to be a necessity for transducing primary cells, is developed. Lentivirus particles, with a set of transgenes encoding an artificial cell-signaling pathway, are used to transform primary T cells as vectors for calibrated synthesis of desired proteins in situ, that is, T-cell biofactory cells. The process is also used to generate primary T cells expressing antigen-specific chimeric antigen receptors, that is, CAR T cells. The two differently engineered primary T cells are expanded and validated for their respective functions, that is, calibrated synthesis of desired proteins upon engaging the target cells, which is specific for the T-cell biofactory cells, and cytolysis of the target cells common to both types of cells. The process is compliant with current Good Manufacturing Practices and can be used to support the scale-up for clinical translation.
开发了一种用于最大化慢病毒颗粒滴度的方法，这被认为是转导原代细胞所必需的。携带一组编码人工细胞信号通路的转基因的慢病毒颗粒，被用作转化原代T细胞的载体，用于原位校准合成所需蛋白质，即T细胞生物工厂细胞。该方法还用于生成表达抗原特异性嵌合抗原受体的原代T细胞，即CAR T细胞。对这两种经过不同工程改造的原代T细胞进行扩增，并验证其各自的功能，即与靶细胞结合后校准合成所需蛋白质（这对T细胞生物工厂细胞具有特异性），以及两种细胞类型共有的靶细胞的细胞溶解作用。该方法符合现行的药品生产质量管理规范，可用于支持临床转化的扩大规模。
