Conventional mass spectrometry is usually limited to masses less than a Megadalton because of heterogeneity. Charge detection mass spectrometry (CDMS) can overcome this limitation and extend accurate mass measurements into the Gigadalton regime. Recent technical advances have dramatically improved the sensitivity of CDMS. Applications of CDMS to the analysis of complex biopharmaceuticals, including gene therapy products and vaccines, will be presented.

Clinical-grade CAR T cell drug products contain a heterogenous mixture of phenotypically and functionally distinct cells. Such heterogeneity necessitates innovative and comprehensive strategies to characterize CAR T cell therapy investigational drug products. Here, we present how high-dimensional single-cell analytics in CAR T manufacturing and beyond can be used to resolve drug product complexity and identify potentially key clinical correlates.

Recombinant adeno-associated virus (rAAV) vectors have been widely used for in vivo gene therapy with commercial products approved by FDA. Charles River Laboratories (CRL) PA Biologics has established a platform approach for rAAV vector genome titer quantification, residual host cell DNA quantification and sizing evaluation, and replication competent virus (rcAAV) detection, by adopting real-time PCR as well as ddPCR technologies and using reference materials available from ATCC or biological materials created at CRL.

Standards are essential to the development and control of biological products. Considering their importance, there is no consensus on the source of a standard, the basis and means of standard qualification, and stability evaluation. This talk will discuss principles and practices related to standards used to report potency of biological products and propose strategies. Those proposals will borrow from practices related to quality by design, highlighting fitness-for-use of a standard.

This discussion will cover standards for regenerative medicine products. Discussion will focus on standards for bioassays, published standards and how to implement these standards be addressed. An overview of current standards under development and how to get involved, as well as a forum for questions will be provided.

Currently the gene therapy space is under utilizing the power of biophysics, specifically Analytical Ultracentrifugation (AUC). Besides the quantitation of empty capsids, AUC can also be used to evaluate the physicochemical properties, higher-order structures, and other pertinent properties. The EMA and FDA are seeing QC release specifications based on AUC, therefore it is paramount that AUC methods are fully realized and meticulously developed.

Viral capsid proteins play an important role in cellular targeting and trafficking as part of the viral infection cycle, and thus changes in the viral capsid protein sequence or post-translational modifications (PTMs) might impact viral targeting and infectivity. We evaluated the role of AAV capsid protein PTMs on AAV transduction potential by generating AAV2 and AAV5 capsid mutants and performing a stress study.

When thermally stressed AAV genomes leak out from their capsids, and do it before proteins melt. However, methods that look only at protein unfolding don’t track DNA, so you’re not getting the whole story. This talk will show Uncle’s insight on AAV stability by monitoring DNA ejection from capsids, and how to look at quantifying encapsulated DNA and particle titer.

An integrated control strategy was developed for a late-phase CAR-T cell therapy product based on our current understanding of the product quality attributes, the manufacturing process, and analytical capability. Based on this comprehensive assessment, the overall risk of drug product to patient safety and product efficacy was determined to be low, demonstrating that the combined operational and testing controls are suitable to ensure product quality from the proposed commercial process.

We used a very common analytical method to characterize subvisible particles in biologics (Micro-Flow Imaging) and applied the technique to cell therapy products. We have developed customized machine learning algorithms specific to cell therapy MFI data for rapid classification of images to establish particle profiles. The method has overwhelming potential to monitor cell particle size distributions, cell debris, cell agglomerates, as well as extrinsic material from batch to batch.

Live bacterial products (LBP) are novel therapeutics with the potential to treat unmet needs in rare genetic diseases, such as phenylketonuria. Viability is a critical aspect of LBPs and traditionally determined through colony-forming unit methods. Another approach to determining viability is automated cell counting. When the methods were compared, the cell counting results were more indicative of in vivo activity of the LBPs and is now used to determine dose.