One challenge is enhancing gene delivery efficiency. 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 Therapeutics, Inc. overcomes these challenges of inefficient gene delivery and low cell viability following electroporation by introducing Quantum Nufect™.
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 utilized 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 ), implicating potential benefits for use in clinical scale gene-modified cell production. 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. The evidence suggest that Quantum Nufect™ is a superior buffer system for generating virus-free CAR-T 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 also facilitates the transfection of (B) commonly used HEK293 cells as well as the hard to transfect C17.3 cell line. (C) When comparing Quantum Nufect™ with commonly used buffer system (Lonza), Quantum Nufect™ is 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 solution (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.