Showcasing a bioassay CMC strategy for a challenging bispecific and the steps taken to bring this molecule to the market. Two checkpoint inhibitor targets on a bispecific which require a suite of bioassays to measure potency. Exploring cell-based functional assays and multiple binding assays including dual-binding to understand benefits/drawbacks of each to build a comprehensive control strategy.

FDCs, a new product class tailored for solid tumours promise many advantages over ADCs including rapid tumour penetration and faster clearance. However, due to the high DAR achievable with our novel technology, new analytical strategies need to be applied as the linker-payload becomes a dominating feature by mass. We’ll share LC-Mass spectrometric, Dynamic Light-Scattering, and Nanofluoresence-temperature unfolding data to show how we discover and characterize novel FDCs using our discovery engine.

We report a novel ultrahigh-throughput screening method called enzyme proximity sequencing (EP-Seq) that accelerates enzyme engineering for cancer therapy. Our method leverages yeast display and deep mutational scanning to characterize the stability and catalytic activity of pooled enzyme variant libraries in a massively parallel fashion. Enzyme activity is read using radical-based proximity labeling and sequencing, which generates training data for machine learning. Our approach will enable accelerated development of highly stable and active enzymes for metabolic cancer therapy.

Adeno-associated virus (AAV) sample is typically composed from full, empty, and partial particles and other impurities. In this talk, the characterizations of AAV samples, including size distribution, capsid composition, capsid modification, and nucleic acid heterogeneity using various biophysical methods like different types of analytical ultracetrifugation, liquid chromatography mass spectrometry (MS), charge detection MS, cryo-electron microscopy, capillary electrophoresis, mass photometry, will be introduced, together with the relation of their properties to biological activities.

In this presentation, we consider therapeutic approaches to limit detrimental effects of extracellular hemin released from hemoglobin and discuss contemporary biophysical methods for the evaluation of hemin-protein interactions with regard to various tasks, from the manufacturing controls of the hemin impurities in blood-derived products to the development of efficient hemin scavengers. This talk provides an overview of the research studies towards therapeutic development for hemolytic disorders, with a focus on biophysical characterization of potential hemin scavengers. 

Cell-based bioassays often face limitations with sample throughput, speed, and automation, and usually rely on indirect readouts utilizing reagents and labels. Here, we present an innovative feasibility study for development of a direct, fast, and label-free MS-based cell assay for therapeutic antibody potency characterization, using automated sample preparation, rapid MALDI-TOF mass spectrometry, and advanced data analysis. The MS cell bioassay provided comparable quantitative results to the routine luminescence-based method.

CEACAMs are membrane-associated glycoproteins that are overexpressed in some tumor types. The antibody-drug conjugate, tusamitamab ravtansine, specifically recognizes the human CEACAM5. To understand the structural basis of this specificity, we mapped the epitope-paratope interface between hCEACAM5 and tusamitamab by various biophysical and biochemical methods and cryogenic electron microscopy. The Cryo-EM structure of the tusamitamabCEACAM5 complex revealed a discontinuous epitope involving residues in the two CEACAM5 domains and an N-linked mannose. This talk will also discuss the structural biology vs. other methods of epitope-paratope interface characterisation. 

MAM is an emerging mass spectrometry-based technique that allows for simultaneous monitoring of multiple quality attributes of therapeutic proteins on an individual amino acid level in a single analysis. This method streamlines biopharmaceutical analysis, improving product and process understanding, reducing resources and accelerating development of complex biologics. We will present the implementation of an automated MAM platform from analytical characterization to process analytics on two robotic liquid handler types and the assessment of cross-site performance of the entire MAM workflow.