In silico tools to rapidly assess physical and chemical liabilities of biologics such as aggregation, oxidation, deamidation will be discussed. Use of these tools for early risk assessment and to identify key critical quality attributes (CQAs) during manufacturing will be demonstrated. In addition, case studies will be discussed to obtain mechanistic understanding of the underlying cause of degradation and address unique stability challenges observed during development.

Advancement in PEGylation technologies has provided new opportunities in drug discovery by enhancing clinically relevant properties of biological molecules. Covalent or non-covalent attachment of polyethylene glycol (PEG) to proteins has been shown to increase half-life, reduce immunogenicity, improve solubility and stability. However, this new format presents new challenges for established analytical methods and controls, especially with respect to controlling charge variants. In this presentation, I will discuss how we overcome these challenges to design appropriate analytical controls to ensure purity and consistency for a novel PEGylated antibody. 

The complexities of emerging nanoparticle-based delivery systems present specific challenges for the development and implementation of effective control strategies in the early and late phase. A fundamental understanding of how biophysical attributes connect with product quality is needed to underpin effective analytical control. This talk will examine how physicochemical characteristics of nanoparticles may be considered together with challenges associated with leveraging instrumental methods for biophysical control.

New modalities present new challenges in their analysis. For example, due to their intrinsic structural complexity and complicated manufacture processes, viral capsids or particles (e.g., adeno associated virus, AAV) are markedly more heterogenous than well-established modalities. In this talk, both new methods and findings (e.g., PTMs) will be presented. Furthermore, I will discuss analytical artifacts, which are common yet under-appreciated, thereby often leading to erroneous interpretation and counterproductive approaches.

During candidate lead selection, hundreds of molecules are screened for potency, binding and developability characteristics, such as hydrophobicity and thermal stability. As high-throughput expression/purification is implemented, analytical technology must adapt to increased number of samples, often with limited material availability. Here, data is presented showing implementation of high-throughput analytical technologies for CE, LC, and biophysical methods to support high-throughput screening processes to support lead selection.

In this talk, we will discuss the novel crystal structure and characterization of human heavy-chain-only antibodies that reveals a novel, stable dimeric structure. Our findings indicate that human heavy-chain dimers can be secreted efficiently in the absence of light chains, may show good physicochemical properties and stability, are structurally similar to Fabs, their formation offer insights into their mechanism of formation, and may be amenable as a novel therapeutic modality.

Nonspecific binding is a common phenomenon that plagues many antibody discovery and optimization campaigns. The ability to rapidly identify and reduce off-target binding early during candidate selection can help improve the odds of selecting lead antibodies with drug-like properties. I will discuss trends in antibody developability from our analysis of clinical and pipeline datasets across antibody modalities, and apply these learnings with a recent case study that utilizes predictive library design and high-throughput developability screening to improve specificity during optimization.

The AQS3pro, powered by Microfluidic Modulation Spectroscopy - A highly precise and automated IR spectroscopy tool, utilizing secondary structure analysis to determine stability, similarity, and structure of proteins and biomolecules.

Co-formulation dosing enables patient convenience and increased compliance due to simple one-vial image and single IV infusion, lowering the administration time. In addition to the combined therapeutic effect of two mAbs, co-formulation also offers the advantage of efficient manufacturing processes. This presentation highlights key challenges and opportunities with case studies for co-formulation composition and process optimization supporting early-stage development with line of sight to late stage.

Formulations for antibodies at a high concentration are required to provide optimal conformational and colloidal stability. DOE methodology is often used to identify the pH, ionic strength, and excipients, followed by real-time assessment of stability. This talk will discuss high throughput techniques to predict conditions for conformational stability and solubility and compare to real-time stability generated for multiple high concentration antibody formulation.”

Formulation development is an important strategic tool that often provides a competitive advantage. Developing robust formulation in a short development time is the need of the hour. Efficient use of prior knowledge and available product understanding are important considerations. An in-house case study will be presented, demonstrating a thorough data mining followed by a detailed evaluation of choice and role of excipients. The case study will highlight the selection of polysorbate concentrations required for optimal product stability.

Interactions between water molecules and APIs in biologics make water protons a sensitive marker of the API structure and organization.  As such, water proton NMR (wNMR) is advantageously used to quantitatively characterize the stability of biopharmaceutical drug products.  Importantly, the analysis could be done quickly and noninvasively, in the original drug product container using low-cost wide-bore compact benchtop NMR spectrometers.  This simple and affordable technology could open new prospects for formulation research, drug product QC and point-of-care applications.

Metal ion impurities are responsible for the chemical degradation of formulation components such as the polysorbates and active protein molecule. The use of metal ion chelators, like EDTA and DTPA, to mitigate chemical degradation is a well-established strategy to tackle this problem. In this work, we assessed the impact of effectiveness of different types and different levels of chelators in mitigating Polysorbate 80 degradation.

In this talk, I will discuss the latest developments in rational strategies to evaluate the effects of surface-mediated stress on antibody instability. The focus will be on early detection and mechanistic understanding that will enable the creation of an effective control strategy in the industrial setting. 

Particle characterization of biologics is challenging especially for investigational drug products that are prone to particle formation during their development, manufacturing, and administration. To address some of the frequently observed challenges during particle characterization of intravenous-admixture compatibility studies, we are proposing a novel approach to characterize subvisible particles using existing tools such as light obscuration and flow imaging microscopy.