Chimeric antigen receptor T-cell (CAR-T) therapy has revolutionized immunotherapy, offering innovative solutions for cancer treatment. Allogeneic CAR-T therapies utilize T cells from healthy donors or such sources as umbilical cord blood and induced pluripotent stem cells, providing scalable, off-the-shelf treatments with broader applicability and reduced waiting times compared with autologous CAR-T therapies. The manufacturing process includes T-cell activation, genetic modification using viral vectors to express CARs, cell expansion, rigorous quality control testing, and cryopreservation for storage and shipment. These therapies demonstrate clinical efficacy in hematologic malignancies, making them cost-effective and accessible to patients who are unsuitable for autologous approaches. In contrast, autologous CAR-T therapies leverage a patient's own T cells to create personalized, highly targeted treatments. Beginning with T-cell collection via apheresis, the process involves activation, genetic engineering to express CARs targeting specific cancer antigens, cell expansion, and thorough quality control. Despite challenges in T-cell quality and manufacturing complexity, autologous CAR-T therapies have shown remarkable success in treating acute lymphoblastic leukemia and non-Hodgkin lymphoma, offering hope to patients who have exhausted other treatment options. Together, both approaches highlight the transformative potential of CAR-T therapies in cancer treatment.
This second part of a two-part article provides a clear understanding of microbiological load reduction during cleaning processes in the non-sterile pharmaceutical manufacturing. Different microbiological test methods are used to get to an outline if the cleaning process can already be seen as a microbiological risk reduction step. Finally, cleaning validation considerations using different cleaning chemistries are discussed in this paper.
A critical function of container closure systems-defined as one vial, one stopper, one seal, or one press-on cap-used for the packaging of injectable drug products is to maintain closure integrity over the life cycle of the drug product when exposed to a range of storage and transportation temperature conditions. Many injectable drugs are packaged in glass vials affixed with an elastomeric closure secured by an aluminum seal using sufficient crimping seal force to obtain the target residual seal force to assure container closure integrity (CCI). And more recently, interest in closures comprised of press-on caps pre-assembled with stoppers. The development of advanced therapy medicinal products (ATMPs), vaccines, insulins, and many biologics requiring extreme cold storage management pose CCI challenges not encountered at ambient and base cold storage temperatures impacted by differences in packaging component coefficients of expansion and elastomer viscoelastic transition to a glass-like state beyond -50 degrees C. ATMPs, including cell, gene, and RNA therapies, are collectively defined as a class of highly innovative novel drugs requiring complex cold chain logistics, driving sustained growth of the cold chain medicine market and a need for injectable primary packaging capable of maintaining CCI at extreme cold storage (-80 degrees C) temperatures. This study evaluates the best-in-class elastomer compounds, formulations, and coatings, affixed with aluminum seals and press-on caps to Type I borosilicate tubular glass vials conforming to the International Organization for Standardization (ISO 8362) injection containers requirements.
Validated cleaning approaches, especially legacy processes, are challenging to modify. To make the case for change, strategies such as the use of digital twins and continuous monitoring enable return-on-investment calculations and real-time quality assessment to mitigate risk. Embracing these strategies is a means to drive out waste and improve the overall cleaning process. The authors explored the elimination of a water rinse and blow down following the caustic wash step, examining the potential safety considerations, the effect on the quality of the cleaning process, and the potential benefit of implementing the change
Pneumococcal infections, stemming from Streptococcus pneumoniae , present a substantial health risk, especially among vulnerable populations. To counter this, vaccines employing bacterial capsular polysaccharide (CPS) have been developed. However, the purification of CPS is a pivotal vaccine manufacturing step, with the clarification process playing a crucial role in achieving optimal product yield and purity. This research focuses on refining the clarification step by assessing the performance of various depth filters. The study explores both conventional cellulose -based and innovative synthetic depth filters, utilizing pneumococcal fermentation harvests from two challenging serotypes. Evaluation criteria include hydraulic performance, throughput, and filtrate turbidity. Employing the constant flow rate sizing method for scale -up, the study determines that synthetic depth filters with suitable pore sizes demonstrate low resistance and high throughput, suggesting their efficiency in impurity removal. Overall, optimizing the clarification step is crucial for improving downstream processing of pneumococcal polysaccharide conjugate vaccines, and this study provides valuable insights into selecting and optimizing depth filters for efficient clarification, ensuring high product quality and process efficiency in vaccine manufacturing.
The harvest step in a monoclonal antibody downstream purification process removes biomass, particulates, and other material from the cell culture to generate a product stream suitable for purification by liquid chromatography. In a widely employed, two-step harvest process, coarse solids and intact cells are first removed using a centrifuge, and the resulting centrate is further clarified by depth and membrane filtration in series. Conversely, direct depth filtration has the potential to greatly simplify the overall harvest process by directly loading the cell culture broth on high -capacity depth filters and consequently removing the need for centrifugation. The authors evaluated this potential of direct filtration for multiple biopharmaceutical candidates to understand the underlying mechanisms that define fouling and the resulting capacity limitations in these filters. Here, the authors present a methodology to assess the contribution of individual filters to overall capacity of the direct filtration train, and elucidate how capacity is affected by characteristics of the cell culture broth and process parameters, including for enhanced upstream processes such as those incorporating high cell density perfusion. The findings highlight the limitations of direct filtration when implemented in large-scale facilities for high cell density processes. Ultimately, increased understanding of the fundamental principles that govern depth and membrane filtration can aid in moving from a trial -and -error to a more predictive and methodologybased approach for harvest development. This article is Part 2 of the study.
Single-use biomanufacturing is a rapidly growing industry. In recent years, single-use centrifuges have become viable alternatives to traditional filters, promising 90% reductions in waste materials and improvements in process time and yield. However, these systems often have several fluid paths with different materials, complicating extractable studies This case study describes how the risk assessment of the first-to-market single-use disc-stack centrifuge was conducted. This study shows how a dramatic modification of the surface-area-to-volume extraction ratio was required for the single-use system and how this was considered during interpretation of the results respecting industry best practice. Furthermore, it's demonstrated how the detection of volatiles can vary depending on the approach of analysis, surface coatings, and where analytical study design should be considered to mitigate missing volatile extractables.
This commentary summarizes the derivation of clonal HEK293 suspension cell lines, selection of clones for rAAV production, and design of experiments-based optimization strategies for characterization of one clonal isolate for high yield rAAV manufacture.
