I will present an end-to-end analytics workflow developed at Large Molecule Research (LMR) at Roche Innovation Center Munich. It allows full automation of sample registration, wet lab processing and MS measurement, as well as data management, analysis and reporting applied on heterogeneous antibody-based drug candidate samples. Data and metadata are ultimately consolidated and stored in a way that enables mining and efficient and informed decision-making.

We present case studies of an innovative microfluidic, nanoliter-scale, and automated immunoassay platform that improves bioanalytical productivity during pharmaceutical development by streamlining workflows, improving data quality, and greatly reducing the consumption of precious reagents and samples in a wide range of applications including product titer and impurities analysis. 

In recent years native MS has gained significant popularity as a tool for intact mass analysis of biopharmaceuticals. Key strengths include excellent spectral data quality and the ability to interface the MS with a broad variety of separation techniques. Here case stories and learnings from the transition to native MS in a biopharmaceutical development lab will be presented.

Many important analytes in biopharmaceutical manufacturing are chemically complex such as culture media and proteins and the use of traditional analytical methods can sometimes be problematic for these materials. Therefore, there is a need for rapid characterization methods for analysing lot-to-lot variance and predicting material/process performance. These materials are intrinsically fluorescent which means using multi-dimensional fluorescence spectroscopy measurements are a viable option for many applications. Here we describe the benefits and operational use of these methods.

Stability of biopharmaceuticals is a complex matrix of parameters that plays a crucial role through the entire product life-cycle. Through case studies, we discuss how Prometheus provides clarity by delivering precise and reliable stability measurements, outperforming existing tools, to inform critical decisions throughout the product life-cycle.

The technical and regulatory developments of the last decade have set the path for a rapidly growing monoclonal antibody biosimilar market. However, the need of ensuring product consistency across manufacturers, jurisdictions and over time has also become apparent. International standards for monoclonal antibodies are publicly available reference preparations that support bioassay performance and calibration of local reference standards. By defining international units of bioactivity, they allow global harmonization and traceability of bioassay data contributing to long-term product consistency.  

Efforts are underway to modernize ICH Q2(R1), the authoritative guidance on analytical method validation, and draft Q14, which will provide guidance on modern concepts in method development. Q2(R2) and Q14 will provide a roadmap for efficient communication between applicants and regulators on the process of taking an analytical method from definition of intended use through development and validation, ultimately resulting in robust, approvable methods and facilitating management throughout their lifecycles.

The complexity of biotherapeutic products and their manufacturing processes yields a variety of impurities, which must be monitored and controlled to minimize safety concerns and ensure product quality. Process-related impurities arise from the manufacturing process and include host cell DNA, host cell protein, and particulates. This presentation will focus on approaches for monitoring impurities, including a discussion of existing USP standards and new standards under development to support impurity testing.

Well-developed and broadly-reactive HCP ELISA are typically used during the purification process to ensure removal of HCP and to demonstrate process consistency and final drug substance purity. Regulatory guidelines require use of orthogonal methods to ensure the HCP ELISA is fit for purpose. AAE and MS have emerged as tools for identifying HCPs that persist through downstream purification processes. These methods complement HCP ELISA and provide a more comprehensive HCP impurities analysis.

Bioprocess variations are caused by raw materials, human interactions and the biological system itself. The resulting impurity profile needs to be identified along the process chain and decisions on how to adjust the control strategy of the subsequent step need to be taken. This contribution aims to show how the complexity of the variations in cell populations and impurity profiles can be identified and reduced by advanced flow cytometry and fingerprint methods. Those methods feed into an integrated process model for holistic control strategies.

Distinct molecular attributes can critically modulate the physicochemical and pharmacological properties of protein biotherapeutics. For a quality by design approach, a detailed understanding of key structural features and degradation pathways is required to enable protein design and CMC development to proceed in a rational manner, whereby quality attributes are understood and appropriately controlled. Due to their highly potent nature, and resultant low dose, toxin-based therapeutics present some unique analytical challenges. Case studies will be presented where insight into quality attributes for toxin therapeutics has been gained using highly sensitive analytical characterisation techniques.

Protein self-interactions are an intrinsic behaviour that can adversely affect the developability of therapeutic proteins. Our recent work on NMR-based interaction & behaviour assessments permits the simultaneous detection of weak protein-protein and protein-excipient interactions. We discuss applications to protein formulation optimisation, de-risking mAb co-formulations, candidate developability assessments as well as residue-level structural details that can guide protein design.

Progress in drug design has led to the development of new peptides, proteins, and drug molecules. However, the limited ability to selectively deliver these molecules at well-defined dosing regimens remains a significant challenge. These challenges can be bypassed by using novel technologies such as 3D Printing/Bioprinting. Various designs with high drug payloads that formulated by 3D printing and characterised using advanced characterisation techniques (e.g. μCT, ToF-SIMS) will be discussed.

Cell line development includes the screening of thousands of clones. The aim is to find the few that are stable, grow as expected, and produce high yields of the bioproduct.  Speed up antibody discovery and development by moving higher quality candidates downstream. How? Carry out expression level analysis label-free in crude samples. Combine it with early-stage glycan characterization both in a high-throughput mode.