Whitepapers

Application Notebook highlighting Liquid Chromatography techniques commonly used for monitoring Critical Quality Attributes (CQAs) in Bioprocessing Applications

Catalent has developed a cutting-edge bioassay using real-time quantitative reverse transcription (RT-qPCR) in a duplex format to measure transcription activity in cells treated with ligands or transgenic vectors. This reliable and reproducible assay is a valuable tool for evaluating the relative potency of various test substances. Enhance your research with Catalent’s robust cell-based potency assays.

Transcriptional activity within a cell can be used to evaluate cell response to a ligand or promoter activity within a transgene or plasmid within a cell. Catalent has developed a relative potency bioassay using real-time quantitative reverse transcription (RT-qPCR) in a duplex format to assess relative transcription activity in cells treated with ligands or transgenic vectors. The assay utilizes two fluorescent dyes with minimally overlapping emission spectra that allow real-time monitoring of the gene expression of both target and normalizer genes. The assay does not require purification of the mRNA produced by the cells once lysis has occurred. Normalizing the qPCR cycle thresholds (CT) of the target transcript to the reference transcript allows response curve to be generated and compared to a reference standard. The generation of a four-parameter fit curve analysis from raw qPCR cycle threshold data allows for comparison of relative potency and assessment of suitability based on curve parallelism. The assay platform has been used by Catalent to qualify a repeatable, accurate, linear, and specific bioassay for assessing relative potency.

The manufacture of protein-based drugs is complex and relies on using biological host systems. This can result in small changes in protein structure during production and formation of protein variants that can have a large impact on functionality. This heterogeneity — variations in the protein size, charge or structure — can significantly impact the safety and activity of the final biotherapeutic or biosimilar therapy, potentially hindering their beneficial effect. It is vital that charged variant profiles of biologics are adequately characterized, as many post-translational modifications (PTMs) may alter the charge of the molecule, in turn impacting its stability, pharmacokinetics and pharmacodynamics. In this article, Catalent explores protein variants, focusing on charged variants, by outlining their impact on protein-based drugs, and explain how specific characterization techniques can be used to determine product safety and efficacy.

Transcriptional activity within a cell can be used to evaluate cell response to a ligand or promoter activity within a transgene or plasmid within a cell. Catalent has developed a relative potency bioassay using real-time quantitative reverse transcription (RT-qPCR) in a duplex format to assess relative transcription activity in cells treated with ligands or transgenic vectors. The assay utilizes two fluorescent dyes with minimally overlapping emission spectra that allow real-time monitoring of the gene expression of both target and normalizer genes. Notably, the assay simplifies the process by eliminating the need for mRNA purification, enabling more efficient and accurate analysis. Normalizing the qPCR cycle thresholds (CT) of the target transcript to the reference transcript allows the response curve to be generated and compared to a reference standard. The generation of a four-parameter fit curve analysis from raw qPCR cycle threshold data allows for the comparison of relative potency and assessment of suitability based on curve parallelism. Catalent has successfully implemented this assay platform to develop a reliable, accurate, and specific bioassay. It stands out for its linear response and reproducibility, making it a valuable tool for evaluating the relative potency of various test substances. Join us to explore how these robust cell-based potency assays can enhance your research and provide critical data on drug product potency.

Biophysical characterization is critical to understand the make-up and behaviors of biologic therapies and vaccines, both early in development and throughout the manufacturing scale-up process. As biologics become more complex in structure, and as scientists improve their understanding of the effects of structure on stability, efficacy, safety, etc., there is a need to develop new and improved analytical methods to characterize biologic products. During this presentation, experts will discuss the latest challenges in biophysical characterization and will present solutions to overcome these challenges.

The characterization and analysis of advanced therapies, such as cell and gene therapies (CGTs) can be difficult, as these products are designed to function using complex mechanisms of action (MOA)s. There are a wide range of challenges associated with accurately assessing the potency and impurity profiles of these complex biologicals. As many CGT programs qualify for accelerated review pathways, novel approaches for analysis and characterization can help generate data that allows for real-time decision making and faster development timelines. Catalent has developed a relative potency bioassay using quantitative polymerase chain reaction (qPCR) to assess relative transcription activity in cells treated with ligands or transgenic vectors. The assay platform can be used to qualify a repeatable, accurate, linear, and specific bioassay for assessing relative potency for CGTs, mRNA- and other nucleic acid-based therapies.

Metabolic Flux Analysis (MFA) is a powerful technique used to characterize metabolic phenotype driving to improved productivity in biomanufacturing. MFA is most powerful when absolute concentrations of the metabolic intermediates are measured, but doing so in practice is often impractical due to inherent limitations of conventional absolute quantitation by mass spectral analysis. Recently, advances in artificial intelligence (AI) have been applied to solving the problem of broad, untargeted, absolute quantitation in liquid chromatography-mass spectrometry (LC-MS). These new approaches extend readily to the determination of absolute concentrations of stable isotopically-labeled metabolic intermediates, offering a new tool for MFA. Dr Sam Yenne from Metalytics Inc., a company that specializes in the science of metabolic flux, has recently assessed and adopted Pyxis, a new tool for absolute quantitation of raw LC-MS data. Dr Yenne describes MFA, its utility in biopharmaceutical development, associated challenges and how Pyxis impacts those challenges.

NGS-Based Viral Safety Testing for ATMPs: Mandatory or Just an Option?

Learn highlights from the 2024 event, including new strategies for viral clearance and NGS applications for viral safety testing.

Cell banking and cell line characterization experts are sharing why HEK293 cells are important for viral vector production for CGT products.

Your biologic can only be as successful as the cells behind them. Learn about our full spectrum of CGMP cell banking and cell line characterization services.

When complexities of developing donor-derived cell therapies are considered, NGS emerges clearly as a better, cost-effective yet regulatory-compliant solution.

Download Part 2 of the DCO₂ White Paper Series and learn the shortcomings of current measurement technologies. As demonstrated in Part 1, Dissolved CO₂ is a Critical Process Parameter (CPP) in biopharma production processes. However, the sensors currently used to measure and monitor this CPP each have significant drawbacks.

Download our comprehensive white paper and gain valuable insights into the critical process parameters and advance sensor technologies that can revolutionize your Biopharma Downstream Process (DSP). Discover strategies and techniques to optimize efficiency, improve product quality, and ensure the safety and efficacy of your biopharmaceutical products.

Learn the latest techniques for improved chromatographic performance and throughput of AAV analysis.

How does PAT apply to the bioreactor? Download our comprehensive white paper and explore what PAT means in Biopharma Upstream processes. Discover cutting-edge solutions to effectively monitor critical process parameters and ensure product quality.

Are you missing sources of measurement error? Measurement of dissolved oxygen (DO) has long been known as a critical control parameter for optimizing cell growth in bioprocesses. Much has been written about different strategies for proper DO control. However, little information has been publicly available regarding optimization of the actual measurement accuracy and reducing common sources of process-related error. Our White Paper aims to divulge these details for the first time.

Collect More Actionable Data and Optimize Yield with Real-Time Process Adjustments. Learn more how Viable Cell Density Monitoring works for your process in Hamilton’s eBook that features must-see examples of real applications using Hamilton viable cell density monitoring (Incyte) and total cell density monitoring (Dencytee).

Cell-based assays (CBAs) are key tools for evaluating the potency of new biologic drugs. Catalent’s transcription-based bioassay simplifies and empowers the testing of new biologics.

Discover how our latest study demonstrates consistent performance across 250mL and 5L bioreactors, ensuring reliable results at any scale. Dive deeper into optimizing your gene therapy processes with scalable, data-driven solutions.

Adeno-associated virus (AAV) is a critical vector in gene therapy, but process robustness and scalability can pose challenges. This whitepaper demonstrates how Culture Biosciences' on-demand fleets of 250mL bioreactors enable large-scale experimentation and real-time data access to optimize upstream processes. Learn how process optimization can increase the fraction of full AAV capsids, accelerating your path to market.

This whitepaper explores how Culture Biosciences' cloud-connected 250mL and 5L bioreactors provide high-fidelity, continuous data streams to deliver accurate cell growth metrics. Learn how automated data analysis and real-time monitoring of key performance metrics like doubling time and production rate enable rapid bioprocess insights.

Culture Biosciences provides cloud-enabled 250mL bioreactors designed for various applications, including mammalian cell cultivation. Accurate transfer of process setpoints is critical to ensuring the success of moving client processes from other systems to Culture’s bioreactors. This whitepaper presents a detailed characterization study of Culture’s mammalian cell bioreactors, highlighting the precision and consistency achieved during process transfer.

This whitepaper explores how power analysis can help determine the optimal sample size for statistically powered clone screening experiments in bioreactors. Learn how to improve key performance indicators such as titer, rate, and yield while accounting for bioprocess variability.

To help provide patients with greater access to safe, effective biological products, the FDA approves biosimilars on an abbreviated pathway. How should you work with the FDA to navigate this pathway?

Maintaining a supply of high-quality product for patients is reliant on the accuracy and completeness of data generated during the development and manufacturing process. Data integrity is critical to ensure that the electronic and paper records we archive and the information they contain retain evidence of the quality of the work performed. This talk discusses data integrity, what it is, and how to implement a robust data integrity program.