Developability assessment evaluates the drug-like properties of lead candidates at the early stage (i.e. discovery), for the potential of being successfully developed into a stable, manufacturable, safe, and efficacious drug product. It is important to carry out this assessment as early as possible in the pre-clinical stage of development to eliminate candidates that do not present a favorable developability profile. This will minimize the risks of costly late-stage failures and avoid potential issues that may occur during the clinical application of a biotherapeutic.
At JOINN Biologics, a panel of small-scale, fast, and predictive tests are used to evaluate therapeutic antibodies’ developability. The developability evaluation comprises in silico analysis as well as a series of in vitro assays for the drug candidates, so to provide information about the integrity, purity, aggregation, thermostability, charge heterogeneity, glycosylation, poly-reactivity, potency, stability over low pH and accelerated storage conditions, and potential PKPD profile.
- Lower liabilities of deamidation, isomerization, oxidation, hydrolysis in variable especially CDR regions from in silico analysis
- Preferred physico-chemical properties by in vitro analysis
- High affinity to target by biosensor
- High purity by capillary electrophoresis
- Level of degradation, aggregation by size exclusion chromatography
- Low heterogeneity by MassSpec
- Zero non-specific binding from poly-reactivity assay
- High stability over extreme conditions. i.e. low pH, high temp
- Preferred glycan profile
- Favorable PKPD profile shown by FcRn binding
- Acceptable viscosity at high concentration
Cell Line Development
Obtaining a highly productive, single cell derived stable cell line is essential for manufacturing biological drug substance. JOINN Biologics’ highly efficient cell line development process can meet such growing needs and accelerated timelines. Using a combination of high through-put imaging, screening, single cell printing technologies and early product analysis, we are able to evaluate every step in the CHO cell line development process. Our optimized workflow demonstrates successful selection of clonal cell lines with high productivity, high quality and monoclonality.
JOINN Biologics utilizes a CHO-GS cell system (SAFC) coupled with state-of-the-art technology platforms for developing stable cell lines that meet the manufacturing needs of therapeutic antibodies and recombinant proteins. With appropriate risk management, our effective processes are able to fulfill the requirements in productivity and timeline
The goal of process development is to establish scalable, robust, and cost-effective processes suitable for the manufacturing (MFG) of biotherapeutics. There are two key parts of the process development: Upstream Process Development and Downstream Process Development. Upstream PD is focused on cell culture to obtain higher productivity, while Downstream PD is focused on efficiently removing impurities and achieving high recovery yield of the products.
1. Upstream Process Development
High unit volume productivity of cell culture processes can be achieved through optimization of cell culture media, feeds, supplements, and bioreactor conditions such as temperature, dissolved oxygen, pH and agitation, using lab scale culture systems including automated bioreactors and design of experiment approaches.
Multiple production processes including fed-batch and perfusion are being developed. The developed processes are then confirmed in pilot scale in PD labs before transferring to manufacturing.
- Productivity and product quality optimization: DOE screening of media, feeds, supplements etc. using Ambr250, up to 12 x 250 mL bioreactors.
- Optimization of bioreactor operation parameters such as temperature, dissolved oxygen, pH, gassing, agitation and studies of robustness in Biostat and Eppendorf stir bioreactors, up to 14 x 2L.
- Process scaleup and confirmation in STR bioreactor, 1 x 50L and 1 x 200L.
- Seed train development in shake flasks, Biostat B Twin CC – RM Rocker 20 L/50 L, Biostat 10L, and STR 50L.
- Development of batch, fed-batch, and perfusion cell culture processes.
- Harvest process development: Continuous centrifugation, depth filtration, and bioburden reduction filtration.
- Process intensification by high density seed train and/or production cell cultures.
2. Downstream Process Development
There are two main elements in downstream process development: chromatography and filtration, which help to recover biotherapeutic products and remove impurities, such as host cell protein, DNA, and potential viruses. Protein aggregates are removed as well resulting in purified and well-behaved protein products.
Our AKTA chromatography systems allow us to explore different purification mechanisms through various protein-resin interactions, such as affinity, ion exchange, hydrophobic and multimodal interactions. We currently have AKTA avant 25, AKTA avant 150, and AKTA pilot 600S on site for purification development. Purification processes developed on the avant platform will be confirmed on the pilot to ensure the smooth, efficient process scale-up.
Various filtrations are performed during downstream process to remove heavily aggregated proteins and host cell debris (depth filtration), potential bacteria (sterile filtration), potential viruses (viral filtration), and to concentrate and formulate the protein products (ultrafiltration and diafiltration). During process development, we will test various membrane filters in order to process and deliver a purified product at the highest cost-efficiency.
Analytics & Formulation
1. Analytics Development
Analytics provides the information for the quality of the product itself, as well as the levels of impurities. By providing sensitive and accurate analytical data, analytics can help upstream development by performing spent media analysis, or downstream purification by analyzing protein products.
JOINN Bio’s analytical platforms include:
2. Formulation Development
Meanwhile, the Analytics department also design and optimize the formulation of the final product so to achieve sound stability of drug products under various shipping and storage conditions. JOINN Bio’s formulation screen typically includes two steps:
Step 1: Screen
Screenings are initially done in 96 well format, using turbidity, thermostability and aggregation as read out
Turbidity by plate reader
Thermo Stability by DSF
Aggregation by light scattering
Step 2: Confirmation
Using viscosity, activity, and aggregation as read out. Samples may need to go through various conditions, such as concentrating (up to 200mg/ml), multiple freezes and thaws, longer-term storage, or continuous shaking
Viscosity by Visometer
Aggregation by HPLC
Activity by Biosensor