Requirements for Lentiviral Vector Production and Quality Control in Cell and Gene Therapy Products

LVs (lentiviral vectors), known for their ability to efficiently transduce both dividing and non-dividing cells and achieve long-lasting transgene expression, have become one of the most widely used gene delivery tools in cell and gene therapy, especially in CAR-T cell therapy and the treatment of genetic diseases. This article aims to provide a comprehensive overview of the requirements for producing lentiviral vectors and ensuring quality control. It reviews the historical development of lentiviral vector systems, including their engineering from HIV-1 and safety improvements through three generations of system upgrades, and compares the technological progress and regulatory frameworks established domestically and internationally. This article systematically elaborates on the mainstream strategies for current lentiviral vector production, including large-scale manufacturing processes based on adherent cells and suspension cells, the advantages and disadvantages of transient transfection versus stable producer cell line establishment, as well as downstream purification and concentration strategies. Special focus is placed on the quality control system for LVs, covering critical quality attributes such as biological activity (titer), purity, safety [sterility, mycoplasma detection, RCL (replication-competent lentivirus), etc.], and physicochemical properties, along with their testing methods and standards. Lastly, future directions are discussed, including innovations in production technology (e.g., stable cell line development, serum-free suspension culture methods), improvements in analytical techniques (e.g., next-generation sequencing, rapid testing methods), and strategies to overcome challenges in large-scale production. The goal is to offer insights that support the clinical translation and industrialization of LVs-based cell and gene therapy products.

CSTR: 32200.14.cjcb.2026.01.0013