Quantum Nufect

Gene-modified cell therapy involves the introduction of genetic materials into cells. Success of gene therapy is highly dependent on the efficiency of cell engineering and the quantity of engineered cells. The “therapeutic genes” can be delivered to a cell’s nucleus using virus or virus-free gene delivery vectors. Virus-free vectors have several advantages over viral vectors as gene delivery vehicles. Despite these advantages of virus-free vectors, there remain challenges that need to be overcome.

One of these challenges is improving efficiency in gene delivery. Electroporation is currently one of the most efficient methods for gene delivery and is commonly used for virus-free gene therapy. However, following electroporation, viability and expansion capacity of cells are markedly reduced. GenomeFrontier’s Quantum Nufect™ overcomes these challenges of inefficient gene delivery and low cell viability following electroporation.

Quantum Nufect™ is a highly efficient and cost-effective universal electroporation buffer system. In conjunction with an electroporator (e.g. Lonza nucleofector™), it can be used to efficiently introduce DNAs, RNAs, or proteins into multiple cell types with minimal reduction in cell viability.

We demonstrated that Quantum Nufect™ facilitates highly efficient vector delivery into several hard-to-transfect cell types, including human primary T cells (see Figure 1 ). We further demonstrated that during CAR-T cell production, Quantum Nufect™ not only preserves the viability of engineered T cells but also enhances cell expansion following electroporation. These evidence suggest potential benefits for using Quantum Nufect™ as a buffer system to manufacture clinical-scaled virus-free gene-modified cells.

Figure 1. Quantum Nufect™ Facilitates Effective Transfection of Cell Lines as well as Primary Cells.

Quantum Nufect™ facilitates transfection of green fluorescent protein (pBRIGHT-GFP)-encoding plasmids into various cell types including human primary (A) mesenchymal stem cells (hMSC), granule neurons and CD8+ T cells (pMax-GFP, see (C)). This buffer system also facilitates the transfection of (B) commonly used HEK293 cells as well as the hard to transfect C17.2 cell line. (C) When comparing Quantum Nufect™ with a commonly used buffer system (Lonza), Quantum Nufect™ was found to deliver markedly higher live cell transfection efficiency. Importantly, Quantum Nufect™ also preserved the viability (i.e. PI- staining) of more cells (96%) compared with cells that were transfected using gold standard transfection buffer system (38%). Viability is expressed as % of PI- cells (gated in purple, R2), and transfection efficiency is expressed as % of GFP+ and PI- cells (gated in green), with respect to the total cell population. Experiments were carried out using Lonza Nucleofector 2B.