Gene therapy

Choosing the best clone for stable viral vector production

GENE THERAPY REQUIRES VIRAL VECTORS IN HIGH AMOUNTS AND AT THE HIGHEST PURITY

Recent developments in genome sequencing technology allow researchers to identify rare diseases with monogenic mutations faster than ever. Effective delivery of gene therapeutics into target cells is a key step for efficient gene therapy and can be achieved by packaging genetic material in viral vectors, such as adeno-associated viruses (AAVs) or lentiviruses (LVs). 

NEXT-GENERATION VIRAL VECTOR MANUFACTURING

The ever-growing demand for next-generation gene therapeutics requires fast and cost-efficient viral vector manufacturing, which still relies on transient transfection of the plasmid into the cell of interest. To overcome batch-to-batch variations, it is critical to develop stable viral vector producer cell lines from single cells and select the most efficient lines for further manufacturing. HEK293 and HEK293T host cell lines are routinely used to produce viral vectors at high titers. Scientists recently developed a rapid method to generate stable producer cell lines from single cells to maximize cell growth for efficient LV production.1

VIRAL VECTOR MANUFACTURING

Our extensive product portfolio offers a range of innovative technologies and instruments that significantly increase the speed, quality and efficiency of your workflows.

SINGLE-CELL CLONING AND MONOCLONALITY ASSURANCE

Our technology dispenses hundreds of single cells within minutes and assures clonality through label-free, image-based cell isolation. The new F.SIGHT™ 2.0 also prevents cell loss through the Cell Focusing feature and allows cell sorting based on fluorescence. Early stage viral vector producer cell line development benefits  from the automation and speed of the new F.SIGHT 2.0. 

The UP.SIGHT™ takes it one step further. Its built-in imager provides double assurance of monoclonality during single-cell isolation and tracks the colonies over time to assess clone recovery and colony growth. The UP.SIGHT is the ideal tool for labs that are looking for an all-in-one solution for clonal cell line generation. Our single-cell dispensers are routinely used for both adherent and suspension HEK293 host cell lines.

HIGH-THROUGHPUT SUSPENSION CULTURE UPSCALING

Growing colonies from single cells in microwells as static cultures requires cells to endure decreased concentrations of nutrients and oxygen in the medium for extended periods of time. Whereas the S.NEST, a high-throughput microbioreactor with CO2 incubator functions, optimizes clonal expansion, adaptation to suspension culture and upscaling. It offers continuous mixing in 96-well/24-well plates for suspension cells by providing a higher diffusion rate of oxygen, which creates a more homogeneous culture environment with real-time monitoring. ​

CONTROLLED ENVIRONMENT FOR UPSTREAM DEVELOPMENT

SOLARIS benchtop bioreactors enable the flexible and scalable expansion of selected clones without introducing variability, which can occur when culturing cells in shake flasks. Our technology allows researchers to actively control parameters such as pH and oxygen via an intuitive software and work with up to 24 units simultaneously. ​

A ROAD MAP TO IMPROVE EARLY DEVELOPMENT EFFICIENCY IN VIRAL VECTOR MANUFACTURING WORKFLOW

Transfection

Single-cell cloning

Early suspension culture

Upscaling

Selection of lines & cell banking

Related products

F.SIGHT 2.0™

UP.SIGHT™

S.NEST™

Solaris Bioreactors

CELLCYTE X™

CASY

References

  1. ​Chen YH, Pallant C, Sampson CJ, et al. Rapid lentiviral vector producer cell line generation using a single DNA construct. Molecular Therapy: Methods & Clinical Development. 2020; 19: 47–57. DOI:10.1016/j.omtm.2020.08.011.