Development and application of an integrated autonomous system for process analytical monitoring of upstream processes producing monoclonal antibodies will be described. Our platform uses automated aseptic sampling and distribution to analytical destinations, including integrated LC-MS for near real-time product quality assessment. Supporting at-line analytics using various MS analyses were also performed for deeper process characterisation. Application for optimisation of mAb galactosylation using a design of experiments approach will be discussed.

Residual host cell proteins (HCPs) can impact patient safety and/or product quality. As the standard ELISA methods are usually not capable of identifying/quantifying single HCPs, LCMS-based methods were established as orthogonal methods over the recent years. But neither the current ELISA nor LCMS methods can selectively identify enzymatically active HCPs (e.g., polysorbate or protein degrading enzymes). Methods combining the enrichment of enzymatically active HCPs and their identification can close this gap. Here, we present a combination of activity-based protein profiling and LCMS analysis, enabling the set-up of targeted MS-based methods to support process development in a subsequent step.

While the multi-attribute method (MAM) has potential to improve the efficiency and specificity of analytical testing, comparison to conventional methods is critical for implementation in QC. Through a cooperative agreement with US FDA, we have evaluated the performance of MAM versus conventional methods in detecting differences between thermally degraded therapeutic proteins from multiple sources. This presentation will share results and an implementation roadmap to facilitate broader adoption of MAM.

• Common developability pitfalls for novel modalities (e.g., bispecifics, ADCs)
• Strategies for mitigating aggregation, instability, and immunogenicity
• Real-world constraints in applying developability frameworks
• Evolving regulatory expectations for developability data

A PS80-oxidised product had been discovered and validated to reveal early signs of PS80 oxidation and quantitively describe the status of PS80 oxidation regardless of the stress condition and incubation time. The accuracy and precision of this PS80 oxidation marker measurement agreed with industry guidelines and therefore can be used to indicate the status of PS80 oxidation and applied to establish the shelf-life to the potential for PS stability.

Viral gene therapy holds great promise for treating complex diseases, but its success depends on producing high-quality viral vectors. Manufacturing these vectors presents challenges, particularly in optimising yield and stability. Here, we delve into physiochemical process conditions that affect AAV particle stability and aggregation, and we provide insights into how additives can improve particle stability and resistance to critical manufacturing process conditions.

Many drugs, particularly novel modalities, face significant formulation and delivery challenges, such as stability, unwanted immunogenicity, and poor biodistribution. By leveraging the principles and structures found in nature, bioinspired drug design and formulation strategies offer a promising approach to address these issues.

Despite their therapeutic potential, bispecific antibodies present additional developability challenges compared to traditional monoclonal antibodies, due to their complex structures. Through case studies, we demonstrate how format changes can exacerbate developability issues for bispecific antibodies, such as their aggregation propensity and polyreactivity. Our findings provide new analytical approaches and perspectives to assess next-generation biologics, in the overall aim of driving their rational development.

Biologics have now evolved from classical mAbs to proteins that are more difficult to analyse, such as therapeutic proteins, multispecifics, novel conjugates, siRNA-and even in the cell and gene therapy space with adeno-associated viruses, for example. With all these new modalities, state-of-the-art analytics must be developed to characterise them in detail, and MS is a major player in this area coupled with diverse liquid chromatography.

N-glycosylation is a critical quality attribute of monoclonal antibodies (MAbs) as it affects binding to Fcγ receptors (FcγR), which impacts the efficacy and safety of MAbs. Surface plasmon resonance (SPR) represents a promising avenue for glycosylation monitoring online of a bioreactor, as SPR biosensors can record MAb-FcγR interactions in real-time and without labelling. Using FcγRIIA/B and a low experimental temperature, we suggest a rapid quantification method for galactosylation and fucosylation.