Formulation and stability assessments are challenged by the short time development for long-term stability data and the complexity of biotherapeutics. To overcome this issue, anticipation through modeling and prediction at a high level of confidence is required. This presentation will use case studies to describe the strength and interest of molecular dynamic and determinist modeling to support the stability/formulation product development from the early stage to marketed products.

During clone selection and process development of bispecific antibodies the quantification and characterization of half antibodies and chain mispairings is of high importance. To assess the amount of half antibodies, SEC, nrCE-SDS and deglycosylated intact MS were applied. To identify the type of half antibodies and wrongly assembled species only MS methods are suited. For the case of swapped light chains a proteolytic digest to release Fab fragments was required.

Antibody-drug conjugates (ADCs) are an emerging new class of biopharmaceuticals being successfully utilized for treatment of various types of cancer. The complexity and heterogeneity of ADCs make product characterization challenging. Therefore, multi-dimensional analytics and understanding the relationship between physicochemical, biophysical, and biological properties of ADCs is crucial for product development. This talk covers front-to-end characterization of ADCs with a focus on structural analysis.    

Adenoviral vector vaccines are challenging study objects due to their complex composition of viral proteins as well as viral and transgene DNA. The biomolecular and biophysical properties of Ad26 vector vaccines are characterized by various methods with the purpose of, for instance, elucidating function-structure, mode of action, degradation pathways and process/product related impurities. In particular, light scattering, liquid chromatography, mass spectrometry, qPCR and ELISA techniques are used.

A risk-based approach is presented to define and justify specifications. It takes advantage of the understanding of structure-function relationships of the analyte molecule and allows the definition of limits that may extend beyond those determined by clinical experience. As appropriate, it serves as a basis for assessing future variability by incorporating worst-case scenarios. In this sense, the basis for designing robust manufacturing processes and facilitating an efficient product life-cycle management.

For biopharmaceutical companies, it is mandatory to analyze the particle content in every liquid preparation. Bionter’s PC100 particle counter goes beyond the limits of light obscuration: The PC100 counts subvisible particles with a non-destructive measurement approach and offers a smart automation workflow.

Digital transformation, from R&D through to manufacturing, streamlines and accelerates critical drug development and manufacturing practices. A QbD approach with PAT can provide full traceability and non-invasive auditing, linking lab to real-time data during scale-up and reducing development costs. The implementation of PAT facilitates products reaching patients faster, to a higher quality, lower cost and with all necessary, regulatory-compliant data and metadata being captured.

A broad range of chromatography, capillary and chip-based separation techniques are now routinely hyphenated to MS for characterization in biopharmaceutical development. Here case studies from the MS 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.

Decay-associated chromatography utilises a novel, label-free optical technique to quantify proteins directly within mixtures in real-time. Applied to chromatographic separations, the elution profiles of two different proteins can be tracked independently without any data-fitting of the elution profile, even when there is complete overlap in the elution of the two proteins. This presentation introduces the technology and explores its application to several analytical challenges associated with developing and manufacturing biologics.

Mass spectrometry is one of the most powerful analytical tools available for qualitative and quantitative analysis of biologics. In this talk, we will discuss the use of MS-based approaches for the comprehensive molecular analysis of biologics. Particular focus will be given to: 1) Multi-Attribute method (MAM) implementation using QTOF-MS instruments, data-analysis challenges and new software developments; 2) HCP identification and quantification using targeted-MS and SWATH-data independent acquisition.

The advent of bi- and multi-specific antibody formats has added to the complexity of mAbs on structural and functional levels. This has significantly increased analytical challenges for release and characterization method development for both bioassays and physico-chemical methods, primarily due to complex architectures and target binding properties. In this talk, we discuss examples of challenges faced during early development phases and propose a number of potential solutions.

Process-related impurities such as HCPs could have a negative impact on the safety and efficacy of medicinal products. Therefore, tight impurity control in the manufacturing process is required in order to guarantee high-quality medicines. Guidance documents for the European and US markets are available to support product licensing. The interpretation of these guidance documents supported by selected case studies will be discussed to highlight the current regulatory developments.

Host cell proteins (HCP’s) are process related impurities that need continuous monitoring for clearance through process purification steps. Development of platform ELISA assays help support program timelines while enabling process teams make data driven decisions regarding the HCP burden across different purification steps. Platform assays are most suitable when established cells lines and processes are used to support multiple programs in early clinical phases. However, it is important to develop platform assays that can support flexibility in process development to best suit program and project needs. This presentation will focus on strategies to develop robust platform assays and assess the suitability of these ELISA assays for early phases of product development.

Cygnus Technologies’ AAE-MS™ method utilizes HCP antibodies from respective HCP ELISA kits to identify and quantitate HCPs in process samples, from upstream harvest to the final drug substance. This data provides unique insight to the HCPs found in purification process samples and allows data driven investigations and decisions to be made when changing assays or modifying the purification process. Case studies for transitioning between HCP ELISAs and identifying changes in HCP profile during a mAb process scale up will be presented.