Formulation development is the experimental design of a buffer and excipient combination that maintains the integrity of the purified protein during routine bioprocessing, storage, handling, and delivery to the patient. Proteins are molecules with individual primary, secondary, tertiary, and quaternary structures that combine to form a unique three dimensional arrangement. These “higher order structures” give each protein its own “personality”. These molecular characteristics will directly impact the formulation development process, at FUJIFILM Diosynth Biotechnologies, we start thinking about the personality of your protein even before the lab work begins.
Our approach to formulation development includes biophysical characterization, mitigating stress-related molecular degradation and minimizing protein self-interactions to optimize solubility protein and stability. Early molecular characterization using biophysical techniques provides increased success in the selection of formulation components and solution conditions that will lead to a stable parenteral drug candidate. Our laboratories are comprised of comprehensive and modern analytical and formulation capabilities and 20 years of CDMO technical expertise to fully characterize and understand your product and process.
Formulation development efficiency is further facilitated by rapid, automated sample preparation and a toolbox of high throughput bioanalytical assays which includes:
- Understanding how to best utilize biophysical assays to understand a protein molecule’s unique characteristics (preformulation)
- Optimizing formulation component selection early in the process by leveraging key biophysical analyses and stress response information
- Performing protein formulation development with high throughput sample preparation and analysis by biophysical and physicochemical methods
Preformulation studies are designed to learn about the protein’s susceptibility to a variety of relevant stresses, develop a baseline biophysical profile for the protein and determine potential degradation pathways of the product, identify stability-indicating assays.
Our science-driven systematic preformulation studies will utilize biophysical characterization techniques such as static/dynamic light scattering, circular dichroism, differential scanning calorimetry, composition-gradient light scattering, particulate analysis and chemical denaturation to provide insight into the protein’s structural personality, stability and propensity for aggregation. The data driven biophysical characterization is also accompanied by utilizing modeling software, where the “hot spots” for potential aggregation or chemical degradation can be predicted.
We examine the biophysical profile of the molecule as a function of protein concentration, pH, freeze/thaw and agitation stress to aid in the selection of the starting formulation conditions (buffers and excipients).
Preformulation will also seek to understand the effect of process and handling steps on the stability of your molecule, for example:
- Effects of Upstream Processing on the molecule intrinsic characteristics (Expression and Harvest)
- Downstream processing conditions (Purification, Chemical Modifications)
- Processing hold steps
- Viral inactivation steps
- UF/DF (buffer exchange/concentration)
Preclinical/Phase I/II formulations can be frozen liquid, refrigerated liquid or lyophilized formulation dependent on intrinsic stability of each product. Frozen liquid formulations are typically the most rapid form used to support early clinical studies. Refrigerated liquid formulations are often preferred if product is stable but require additional buffer/excipient screening and real time stability at the actual storage temperature to demonstrate physicochemical stability. Lyophilized formulations can be used for proteins that may not be stable in liquid state.
Through developing an understanding of the thermal, physical, chemical, and conformational personality of the protein during preformulation, a rational design approach is used to develop a formulation for preclinical and early clinical studies. Our design approach includes evaluating factors such as
- Effect of pH, buffer type, and ionic strength on conformational stability
- Predictive solubility using Composition Gradient MALS (CG-MALS)
- Effect of excipients ability to increase solubility and stability
- Impact of stress through forced degradation studies
- Accelerated stability
Based on the inherent knowledge of the protein, selection of pH/ionic strength/buffer/excipient screening may vary from selecting formulation components found in approved/commercial mAb biotherapeutics (e.g., for mAb formulation development) to de novo design incorporating preformulation knowledge to guide a statistical design-of-experimental (DOE) design to identify the effects and interactions of the formulation components as candidate formulations are selected. Regardless of the approach, FDB performs protein formulation design with robotic high throughput sample preparation to ensure robust and reproducible development.
The data driven formulation development studies will propose formulation candidates. Real time and accelerated stability studies then identify the recommended formulation. Arriving at a well understood and characterized formulation, our approaches help create an “insurance policy” for the successful clinical manufacturing campaign of your product.
As the clinical lifecycle progresses through Phase III and process validation, additional formulation development is typically performed. The following elements may need to be considered in developing the commercial presentation of the biotherapeutic product.
- Buffer optimization and robustness studies
- A change in the dosing schedule or dosage administration requiring a higher concentration formulation
- Container closure/device compatibility study
- Mock Shipping
- A change in the route of delivery (intravenous, subcutaneous, intramuscular, etc.)
- Introducing a delivery device (e.g., prefilled syringe)
Our formulation development services can help will develop a suitable liquid or lyophilized formulation for your drug substance or drug product to ensure stability of your molecule during preclinical, clinical or commercial use.
Our formulation services include:
- Formulation development and optimization (liquid and lyophilized using SMART Freeze Dryer Technology) of Final Drug Product
- High concentration Formulations
- Standard approach to Monoclonal Antibody Formulation to reduce timelines and costs
- Specialized Formulation Development for Product comparability (including biosimilar comparability)
- Short and Long Term Storage Conditions (Freeze/Thaw Cycles)
- Forced degradation programs (including identification of stability indicating assays and multi-lot comparison)
- Design and execution of drug delivery studies including Device Compatibility and reconstitution
To learn understand your molecules personality we apply our comprehensive Biophysical Characterization ToolBox which includes:
- Differential Scanning Calorimetry – MicroCal VP-Capillary DSC Units: Used in the evaluation of conformational/thermal stability
- Circular Dichroism – Jasco J810: Used in the evaluation of secondary & tertiary structures
- MALS – Wyatt DAWN ® HELEOS ™ II MALS/Calypso GC-MALS/Optilab ® T-rEX ™: Used to to determine molar mass, higher order structure
- Fluorescence Spectroscopy – Perkin Elmer LS55: Used in the evaluation of tertiary structures
- Dynamic Light Scattering & Zeta Potential – Malvern ZetaSizer Nano ZS: Used in determination of Zeta Potential, aggregates
- Particulate Analysis (visible/subvisible) – HIAC 9703+, High Throughput Micro-Flow Imaging ™
- Isothermal Chemical Denaturation (ICD) – AVIA Biosystems: Used to determine protein stability at room temperature or experimental temperature
Our extensive physicochemical methods include:
- Chromatography: size-exclusion, ion-exchange, reversed-phase, hydrophobic interaction
- Electrophoresis: iCE, CE-SDS, SDS-PAGE, microfluidic capillary electrophoresis
- Mass Spectrometry: intact MS, primary sequence and modifications by LC/MSE
- Potency: binding ELISA, enzymatic activity, cell-based assays
- Glycan Characterization