CAR T cells are engineered T cells expressing a chimeric antigen receptor (CAR) on the cell surface. CARs allow the T cell to engage an antigen on tumor cells and activate downstream signaling that leads to destruction of the tumor cell. Surface expression of the CAR is critical for therapeutic efficacy, but often differences in efficacy are observed between constructs that show similar surface expression. We developed a workflow using immunoprecipitation of the CAR from the cell surface to characterize post-translational modifications by mass spectrometry (IP-MS) to gain insights on differences in efficacy.

Cell-based gene therapy drug products require comprehensive analytical assays to characterize their safety and efficacy. This talk will focus on the GMP release tests implemented across multiple phase I/II and III clinical trials and the additional non-GMP characterization assays performed to address the heterogeneity of drug products. Correlative analyses between the results of different analytical assays will be discussed.

Cell and gene therapy processes and products are fundamentally complex. As cell-based therapies continue to be translated and advance through clinical trials, long-term success may hinge on our ability to gain greater process and product understanding through enhanced analytics. For cell-based therapies, the starting source material is a vital component to ensuring therapeutic success. The starting material is inherently variable which directly impacts downstream process development and manufacturing. More and more in-process analytical techniques are being developed and implemented to gain better process and product understanding to reduce risks of potential delays or translational success. This presentation will focus on donor-derived starting materials and the impact of characterization on process analytics, consistency, and overall product quality. 

In all biological products, distinguishing aggregated API from other particle types matters for understanding the root cause of instability. Until now, subvisible particle characterization methods have been unreliable, slow, and difficult to use across many workflows. Introducing the Aura, a 96-well, low-volume, high throughput aggregate and particle imaging system can rapidly size, count, and characterize biological particles and identify them as proteins, non-proteins, cellular aggregates, or other types of molecules.

 The increased utilization of cells in biomanufacturing and as therapeutic products over the last decade has prompted the development and publication of two ISO Cell Counting Standards, ISO 20391 – 1:2018 and ISO 20391 – 2:2019. This talk will discuss 6 success factors for obtaining high-quality cell counting results based on ISO standards, with specific recommendations for the cell therapy manufacturing environment. 

CAR T drug products generally use commercial formulation media with complex, undisclosed stoichiometry of ingredients. The proprietary nature of the commercial formulation limits complete understanding of the impact of formulation excipients on product quality, safety and efficacy. In this talk, we present our formulation efforts to develop defined formulations for T and NK cells and also discuss the ability of formulation excipients to increase the product quality and recovery administration.

Natural killer (NK) cell infiltration into and anti-tumor immunity against solid tumors is often low. Functional and metabolic impairment of NK cells is induced by the suppressive microenvironment of solid tumors due to, among others, hypoxia, metabolites, such as adenosine, and the expression of inhibitory NK checkpoints. Here, we discuss our work in redirecting NK cells to overcome immunosuppressive solid tumor by genetically rewiring their functional and immunometabolic responses.

Lentivirus is a very common tool in cell biology, especially in immunology and cell therapy, acting as an effective strategy for cell editing. However, for some challenging cells (ex: T & B), there are still some struggles to achieve high transduction efficiency and insert the targets into genome stably. At GenScript, our proprietary lentivirus system provides intact and functional viruses customized according to research needs and accelerate drug discovery process.

Plasmids are starting materials consisting of double-stranded, circular DNA and used to produce viral vectors in cell and gene therapy modalities. While the regulatory classification of plasmids for cell therapy has evolved in the recent years, necessary controls for plasmids remain to be an area of continued debate and definition. This talk will cover the key considerations and elements of control strategy for plasmids used in cell therapy manufacturing.  

Lentiviral vectors are largely used as gene delivery vehicles for the generation of gene-modified cell and gene therapies for ex vivo treatment of cancers and rare diseases and for some in vivo applications. Manufacturing of lentiviral vectors and derived lentiviral-based cell therapies is complex and requires rapid and reliable analytical assays to measure product’s critical quality attributes and demonstrate consistent manufacturing, safety, and efficacy. Cell and Gene Therapy Catapult has developed a portfolio of assays for LV product characterisation. This presentation will showcase some examples of novel analytical solutions for the characterisation of LV-based products, which aim to reduce variability and turnaround time.

The purpose of this talk will be to walk through the design and build-out of a multi-product GMP manufacturing facility in an academic setting. The talk would review both the technical design considerations as well as business components to ensure staffing and Phase I compliance.

Cellino’s platform combines label-free imaging and high-speed laser editing with machine learning to automate cell reprogramming, expansion, and differentiation in a closed cassette format, enabling thousands of patient samples to be processed in parallel in a single facility.

Adeno-associated virus (AAV) has emerged as the leading platform for gene therapy; however, challenges exist in scaling up production to treat larger patient populations. The AAV-MAX System addresses key challenges by enabling scalable, high titer, and cost-effective production of AAV.  Here, we will discuss data on the use of AAV-MAX for production of AAV across multiple scales in shake flasks and bioreactors, as well as downstream purification strategies and analytics.