Cell Therapy Analytical Development
Introduction
Therapies involving genetically modified T Cells (CAR-T) are on the frontiers of cancer treatment. CAR-T therapies are innovative, allowing personalization of treatment to a patient’s disease phenotype. While cell therapies show promise to become an effective medical modality, production of CAR-T therapies is challenging. Cells are living products and can evolve in culture. Establishing stringent guidelines to characterize potential therapies is important for success in production. Flow cytometry is an essential tool for understanding the suitability of candidates for cell therapy; however, inconsistencies in cytometric assay performance can be attributed to variability between operators or sites where assays are performed. Automating assay development, cell analysis, and culture health measurements makes expansion and production of cell therapies easier, enabling a quick turnaround from process development to patient.
Automated Flow Cytometric Assay Development
Flow cytometric assays are often used to validate cell lines for their suitability as therapeutics. Proper understanding of the strengths and limits of a developed assay is essential for validating cell therapies. Release criteria for assays in development can include:
- Sensitivity of detection
- Accuracy and analytical specificity
- Precision (extending to assay reproducibility)
Establishing a set of standards by which to judge potential therapies is critical for quality control during scale-up and production. The diversity of potential cell therapies makes designing assays that fit the product important. Even if robust assays are developed and validated, variation between laboratories, operators, and reagents used can cause irregularities in data and compromise assay results. Researchers can mitigate the variation from these methods by automating flow cytometry assays – i.e., by integrating a flow cytometer with an automated liquid handler, allowing for hands-off assay development. Dried, shelf-stable reagents help to prevent inconsistencies introduced by reagent handling or lot variation. Custom reagents can be created to help tailor an assay to a lab’s specific needs. Flow cytometric assays developed can be directly transferred for quality control in scale-up of therapies or in a production environment.
Flow Cytometric Analysis
Quality control of cell therapies must occur before release for therapeutic use. Assays developed are used to determine suitability of cell lines for therapeutic use, using cytometric analyses to measure:
- Cell phenotype
- Cell health and count
- CAR expression
Standardizing characterization assays is difficult. Assays may have been developed using different instruments or reagents, and variability between associates can produce subjective results. Performing validation assays is typically not a plug-and-play process. However, the variability inherent in cytometric assays can be minimized by emphasizing consistency in instruments, reagents and software used. Beckman Coulter Life Sciences offers a one-stop solution to performing validation and quality control assays. Automating cell handling and cytometric analyses offers a solution to variability between associates or across sites.
Cell Characterization and Metabolite Analysis
Cell lines used in production facilities may be grown in huge cultures or bioreactors, making characterization of culture health and media composition difficult. Maintaining ideal culture conditions is essential for producing quality and effective therapies. Automation of cell sampling and staining procedures can quickly return data on cell count and viability, giving valuable insights into culture health and maturity. While essential measurements, these metrics cannot give insight into cell activation state or readiness for harvest. Data on cell volume or culture metabolites gives researchers full insight into the status of valuable therapeutics.
Conclusion
Flow cytometry doesn’t have to be a method requiring oversight and constant revalidation of instruments and reagents. Beckman Coulter Life Sciences offers instrument solutions that integrate together to streamline your analyses, and dried reagents for consistency and stabilization. Implementing these approaches in your existing assay development and therapy validation workflows can improve confidence in your data, and reduce the burden, cost, and time required for using this powerful technique in your cell therapy development and production.
