Introduction

To meet the increasing demand for lentiviral vectors in cell and gene therapy trials, a reliable and scalable manufacturing solution is essential. EurekaBio has developed the EuLV lentiviral vector stable producer cell line production system（hereinafter referred to as “EuLV™ System"）, a suspension-mode, inducible lentiviral vector production system to address this challenge. The EuLV™ system eliminates the need for plasmid transfection. All the necessary packaging genes (VSV-G, gag/pol, and rev), as well as the gene of interest (GOI), are stably integrated into the genome of HEK293T cells to create a stable producer cell line.
In this case study, we used the EuLV™ system to develop an AntiCD19-CAR stable producer cell line. Additionally, we established a high-density culture process in chemically defined medium with low uninduced leakage of producer cells. The results of this study serve as proof of concept, demonstrating that the EuLV™ system can produce high-titer and high-quality lentiviral vectors.

EurekaBio Solution

The following flowchart illustrates the process of constructing and developing a stable producer cell line using the EuLV™ system:

1. Integration of VSV-G, gag/pol, and rev genes: These genes are stably integrated into HEK293T cells to create an optimal packaging cell line. This step involves monoclonal screening to select the best packaging cells.
2. Integration of viral genome transcription cassette: The lentiviral genome transcription cassette, carrying the target GOI gene, is integrated into the packaging cells. Multiple rounds of monoclonal screening are conducted to obtain the optimal producer cells.