Proteins constitute a large and highly variable group of biological molecules, central to all known biological systems. Proteins regulate chemical reactions, provide structural support, control physiological responses and mediate immune responses. This ability to impact almost every aspect of biological processes has made them the focus of the Biopharmaceutical industry.
The range of Biopharmaceutical products has expanded greatly and now includes such products as hormones, enzymes, growth factors, immune signalling factors and antibodies. While proteins are a highly valuable source of potential therapeutic agents, their dynamic and complex nature means the development, manufacture and commercialisation of Biopharmaceutical products remains a challenging process.
Proteins are dynamic structures and often exist in a state of flux between the native and partially unfolded intermediates. Additionally, they have evolved to function in specific environmental conditions. These characteristics provide major challenges for the manufacture and long term shelf-life of Biopharmaceuticals.
The aim of drug development is to design a formulation and a manufacturing process that ensures the protein is able to withstand these stresses, while also meeting the shelf life stipulated in the Quality Target Product Profile. Understanding degradation pathways, detecting and characterizing aggregate formation and detecting changes in structure and conformation are of great value during Biopharmaceutical development, and are central to the QbD (Quality by Design) and Design Space approaches currently being promoted by regulatory agencies.
The majority of Biopharmaceuticals are administered by parenteral routes. To ensure low injections volumes Biopharmaceuticals often contain high protein concentrations. At high concentrations, solution viscosity can become a complication by reducing injectability. Therefore, it is important to screen solution viscosity during formulation development.
A growing trend is the use of pre-filled syringes, which contain silicone oil as a lubricant to facilitate the injection process. However, the presence of silicone oil droplets can impact particle characterization, while also promoting aggregate formation. Therefore, it is important to evaluate of potential impact of these and other contaminants, on the active protein and the patient.
The consequences of protein aggregation can vary, but often include loss of efficacy, reduced shelf-life and immunogenicity. Protein oligomerization will also affect activity and drug efficacy. Due to the impact on patient safety, immunogenicity is the most significant impact of aggregation and is major concern of regulatory agencies. Therefore, it is important for aggregated species, including soluble aggregates and subvisible particles, to be fully characterized using the Archimedes and/or Nanoparticle Tracking Analysis (NTA).
Understanding the progression and characteristics of these degradation pathways is an important aspect of product knowledge that can aid drug development and facilitate problem solving. This requires the coupling of protein aggregate detection with characterizing protein structural attributes.
Whether in early stage pre-formulation screening or a legacy commercial Biotherapeutic, product knowledge is central to optimising stability, manufacturability and problem resolution. Our solutions can be applied to many of the problems associated with the development and commercialisation of protein therapeutics.
We can characterize proteins from monomers up to 10µm subvisible particles:
- The determination of protein molecular weight using SEC-MALS as well as protein oligomerization and % aggregation
- Protein size distribution from 0.3nm to 10µm using Dynamic Light Scattering (DLS).
- Particle size and number distributions from 10nm to 5µm using Nanoparticle Tracking Analysis (NTA) and RMM.
- Provide real time protein size distribution data for Bioprocessing monitoring as a PAT tool.
- Our resonant mass measurement enables distinction between protein and non-protein particles.
We have a range of High-throughput Screening tools to support early stage formulation and candidate screening.
- Protein size distributions can be obtained on both 96/384-well plates using the Zetasizer APS.
- While both protein monomer size and formulation viscosity can be determined using the Viscosizer TD.
- Protein-protein interactions can be measured, characterized and screened using isothermal titration calorimetry.
- Automated protein stability studies using differential scanning calorimeter aids protein engineering, pre-formulation and process development.
Our advanced protein characterization tool set enable detailed studies of;
- Protein-protein interaction affinities and energetics.
- Protein zeta potential and surface charge.
- Protein conformation and protein structure.
- Protein molecular weight, oligomerization and aggregation
These characteristics are central to the understanding of protein function, predicting / assessing stability and characterizing degradation pathways. Such knowledge can support the design of formulations and Bioprocessing parameters to ensure maximum stability and shelf-life.