Charles River Launches New Gene Therapy Manufacturing with Lentivation™ Platform; Dramatically Shortens Manufacturing Timelines

Charles River Laboratories International, Inc., a global leader in providing essential products and services for pharmaceutical, biotechnology, and research organizations, has unveiled its new lentiviral vector (LVV) manufacturing platform, Lentivation™. This innovative platform is poised to potentially transform the landscape of gene and gene-modified cell therapies by significantly reducing manufacturing timelines.
Traditionally, LVV manufacturing for gene therapies has been a lengthy and complex process, often taking up to 18 months. However, Charles River’s Lentivation platform is poised to revolutionize this by streamlining the process and reducing timelines by up to 60 percent. With Lentivation, gene and gene-modified cell therapies can now be manufactured in less than seven months, a significant advancement that could accelerate the availability of life-changing treatments.
Lentiviral vectors are powerful tools in gene therapy due to their unique biology and capabilities. Lentiviruses, a type of retrovirus, possess a single-stranded RNA genome with a reverse transcriptase enzyme. They also feature a viral envelope with glycoproteins that facilitate attachment to host cell membranes. Upon entering a host cell, lentiviruses utilize a reverse transcriptase molecule to transcribe their genetic material. This genetic material contains RNA sequences that code for specific proteins responsible for incorporating viral sequences into the host cell’s genome. What sets lentiviruses apart is their ability to infect both dividing and non-dividing cells, making them suitable for gene therapy applications involving cell types that do not typically divide in adult organisms, such as neurons. This unique feature broadens the potential applications of lentiviral gene therapy, especially for conditions affecting non-dividing cell types. The U.S. Food and Drugs Administration (FDA) and the European Medicines Agency (EMA) have approved several gene therapies during recent years that use different viral vectors and lentiviral vectors have grown in popularity.
The production of lentiviral vectors involves several essential steps yet time consuming steps. First, a packaging cell line (PCL) is cultured and expanded to achieve a sufficient cell population. The HEK293 cell line is commonly used due to its transfectability and adaptability to various culture strategies, making it ideal for lentiviral vector (LV) production.
Next, plasmids are transfected into the packaging cells to generate vector particles. To prevent recombination, plasmids are divided into separate transcription units, requiring the transfection of multiple plasmids in the same packaging cell to express all necessary viral sequences efficiently. Transient transfection is the simpler method to obtain viral particles, but establishing a stable PCL capable of continuous vector production is possible, though time-consuming.
After developing the transfected PCL, cell population expansion is necessary to produce an adequate number of viral particles. LV titers, crucial for assessing vector potency, depend on the titration method used. Titer estimation involves measuring transducing units (TUs), which are viral particles capable of efficiently infecting cells. Flow cytometry and quantitative polymerase chain reaction (qPCR) are common methods for TU quantification. Standard controls are employed to standardize titers across experiments.
Finally, before collection, a purification and filtration step is vital to ensure product safety. The use of partially purified batches in animal studies or clinical trials can potentially lead to toxicity or immunogenicity in patients, emphasizing the importance of this purification step.
Lentivation leverages Charles River’s extensive experience and expertise in LVV manufacturing. The platform combines a robust screening toolbox, off-the-shelf LVV packaging plasmids, phase-appropriate production facilities, and in-house analytics to deliver efficient and high-quality LVV production. This approach significantly reduces process development time and costs while ensuring the highest standards of purity and potency.
The launch of Lentivation comes on the heels of Charles River’s introduction of OTS LVV packaging and AAV pHelper plasmids, as well as the eXpDNA™ plasmid manufacturing platform. These innovations are designed to support LVV-based and AAV-based gene therapy programs, marking Charles River’s commitment to advancing cell and gene therapy manufacturing.
The Lentivation platform will be presented during the BioInsight’s Expert Roundtable: Exploring current and future trends in lentiviral vector manufacturing on September 27, 2023, featuring Ramin Baghirzade, PhD, Senior Director, Global Head Commercial, Gene Therapy CDMO Services.
Beth Tebeau, Site Director, Rockville, MD, Charles River: “Charles River’s LVV platform provides a streamlined, reliable path for therapeutic developers to clinic and commercial. The proven platform builds on Charles River’s established capabilities and processes, fine-tuned over decades spent successfully supporting advanced therapy clients.”