Radically transform your gene editing workflow

In this talk, Tom Quinn presents an innovative approach to enhance the ex vivo engineering of human T cells by integrating cell activation and lentiviral transduction into a single, efficient step, reducing time and resource requirements while maintaining high performance. The Lenti-X T-Cell Transduction Sponge provides a faster, more user-friendly workflow to obtain high transduction efficiency with T cells.

With over 130 clinical trials and eight approved gene therapy products, AAV has emerged as one of the most reliable and versatile tools for delivering therapeutic DNA directly into living organisms. The AAV expression system offers several advantages, including efficient transduction, broad tropism, stable high yields, and compatibility with gene-editing components. Quantitative real-time polymerase chain reaction (ddPCR or qPCR), ELISA, cell-based infectious unit assays, and transmission electron microscopy (TEM) have all been used to determine AAV titers. However, these methods are time-consuming and labor-intensive. In this work, Tom Quinn presents an iOS and Android-compatible smartphone application that analyzes a serotype-specific lateral flow assay and delivers particle number values in 10 minutes. The simplicity of the assay facilitates easy monitoring and optimization of AAV production processes to ensure consistency and confidence in downstream applications.

Adeno-associated viruses (AAVs) serve as essential vectors in gene therapy and biomedical research, requiring precise and efficient titer quantification methods to ensure experimental consistency and therapeutic success. In this poster, Yi Zhao presents a single-wash, automation-friendly Enzyme-Linked Immunosorbent Assay (ELISA) protocol for rapid, reproducible quantitation of AAV titers for serotypes 2, 8, and 9. The single-wash approach minimizes hands-on time and procedural complexity, taking only 90 minutes to complete, which is significantly less time than conventional ELISA methods. Validation studies demonstrated a high degree of concordance with quantitative PCR (qPCR) titer data, underscoring its reliability. Its optimized protocol and validated accuracy make it a valuable tool for advancing AAV-based gene therapy research and production, addressing the field's growing demand for efficient and reliable methodologies.

Efficient T-cell engineering is crucial for the success of CAR T-cell therapy but requires multiple labor-intensive steps, including T-cell isolation, activation, and transduction. Herein, we present a streamlined approach to enhance the ex-vivo engineering of human T cells by integrating cell activation and lentiviral transduction into a single, efficient step, reducing time and resource requirements while maintaining high performance—the Lenti-X T-Cell Transduction Sponge. Our approach leverages a lyophilized alginate-based sponge that is embedded with activation reagents, including anti-CD3 and anti-CD28 antibodies, as well as interleukin-2. The hygroscopic nature and 20–300 nm pore size of the sponge spatially constrain cells and virus to a small area, facilitating efficient transduction and ensuring uniform exposure to activation signals, eliminating the need for spinoculation. The release of activated and transduced cells is facilitated by a chelating release buffer that depolymerizes the sponge, ensuring the rapid recovery of high-quality transduced cells with minimal handling. Our data across multiple donors and operators consistently indicate strong activation and transduction efficiencies, with no adverse effects on growth rates, cell phenotypes, or exhaustion marker expression compared to standard protocols. By providing a streamlined and user-friendly system for T-cell engineering, we strive to drive progress in T-cell therapies and expand their availability to a wider range of patients.