Recent years have seen an explosion in the number of separation techniques, that is hyphenated to MS for routine characterization of biopharmaceuticals and other macromolecules. Here case studies from the biopharmaceutical development lab at Symphogen will be presented. The case studies illustrate the strength and complementarity of using different separation techniques coupled to MS for mapping main degradation pathways and critical quality attributes of biopharmaceuticals in development.

Biotherapeutics offer exciting advances in modulating disease-relevant biology. Many biotherapeutics utilize non-traditional scaffolds and new modalities (cytokines, ADCs, multi-specific mAbs); however, their molecular characterization, agnostic of scaffold, remains difficult. We will demonstrate a NMR-based methodology, which does not require isotope labeling, and can quantify the hydrodynamic/solution properties of biotherapeutics. This approach can be applied to a myriad of biotherapeutic topologies.

Recent advances in separation sciences and mass spectrometry paved the way for the comprehensive LC/CE-MS characterization of therapeutic proteins on the intact native level. Such methods excel through simplicity and low analysis time, while also providing comprehensive insight into the heterogeneity of the target biotherapeutic. Here we will explore several native analysis techniques and highlight their potential for the characterization of critical quality attributes of monoclonal antibodies and related products.

What business intelligence solutions do for leveraging company data, Seeq does for leveraging enterprise-wide process manufacturing data. Seeq enables employees who can derive value from the data to access harmonized time series as well as tabular data from all production areas, visualize, cleanse, quantify and model it very quickly thanks to appropriate analysis tools.

This talk will provide an overview of liabilities that are to be expected for non-IgG based therapeutic proteins. Strategic considerations to mitigate these challenges at the right moment (modified developability assessment concept) will be detailed. Examples from internal projects will be given.

Removal of host cell proteins (HCPs) during downstream processing of biotherapeutic products remains a continuing challenge. The current standard method to quantify HCPs is based on ELISA. However, this approach does not always provide sufficient information. Therefore, the use of orthogonal methods like liquid chromatography-mass spectrometry (LC-MS) has been established. To facilitate high-throughput and reproducibility, we implemented an automated sample preparation workflow, thereby allowing for efficient monitoring of HCPs.

A broadly reactive and well qualified HCP ELISA is critical for monitoring purification process consistency and final drug substance purity.  Here we present an overview of assay qualification steps and demonstrate how the orthogonal antibody affinity extraction and mass spectrometry methods assess ELISA Ab coverage to HCPs present in a given process and identify process-specific HCPs that may co-purify with a drug substance.

Protein stability is critical for the successful development of non-aggregating biopharmaceuticals. We have combined biophysical analyses, protein engineering, formulation screening, and molecular modelling, to characterise factors that influence protein aggregation. Examples will be presented from small-angle x-ray scattering, hydrogen-deuterium exchange mass spectrometry, and molecular dynamics simulations.  Insights are being used to develop improved protein engineering and formulation design strategies for the minimisation of aggregation in liquid and freeze-dried forms.

In gene therapy and vaccine development, a broad portfolio of analytical tools for decision making is key. We use transmission electron microscopy (TEM) for direct sample visualization, where information regarding morphology, integrity, and purity is analyzed. The ratios of filled versus empty viral vectors can be quantified by this method. Our GMP certified TEM services address questions that arise during the production process, facilitating decision making in viral vector development.

Pharmaceutical and biotech companies must innovate to bring not only new and efficacious therapeutic products to market, but also ensure that these therapies meet the growing needs of healthcare providers and their patients. The increasingly competitive patient-centric market puts greater emphasis on product differentiation, including convenient combination products and coformulations. This talk will describe several case studies around the use of an innovative formulation technology in developing such product concepts.

Progress in drug design has led to the development of new peptides, proteins, and drug molecules. However, the limited ability to deliver selectively these molecules at well-defined dosing regimens remains a significant challenge. Thus, this talk will focus on how we move from the current treatment approaches to an innovative one by using delivery devices that manufactured by emerging technologies (e.g. microfluidics and bioprinting).

The increasing numbers of biotherapeutic Monoclonal Antibody products and their biosimilars highlights new regulatory challenges. The roles of the manufacturer’s reference standards during product development and control, and the reference medicinal product in the demonstration of biosimilarity, are widely acknowledged. However, the role of International Standards in a multi-product market is often confounded. Data from the international collaborative study for trastuzumab shows how these preparations can promote product consistency overtime.

Efforts continue to modernize ICH Q2(R1), the authoritative guidance on analytical method validation, and to finalize Q14, which will provide guidance on modern concepts in method development. Q2(R2) and Q14 are intended to 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. So, where are we now and what can we expect?

Genetic stability characterisation of viral vector-based vaccines is a regulatory expectation. Here we show how the genetic stability of such a vaccine was addressed from first-in-human to PPQ including details regarding qualification of a reduced-scale model, critical quality attribute selection and analytical method selection. The results were used to demonstrate the genetic stability of the seed and its suitability to consistently produce a safe and efficacious product.

New analytical technologies like Multi-attribute Methods (MAM) and capillary electrophoresis-mass spectrometry (CE-MS) are being adopted for characterization and quality control of biotherapeutics due to their potential to replace multiple analytical tests. This presentation will focus on characterization of USP mAb reference standards using both traditional and new analytical technologies and discuss considerations to enhance robustness of these emerging techniques.