WuXi Biologics
Offering End-to-End Solutions
- About Us
About Us - Services & SolutionsAbout Us
- CareersAbout Us
- InvestorsAbout Us
- News & ResourcesAbout Us
Case Studies
Case studies for development services
- Developability Assessment
- Improving Product Quality with WuXiaTM AI Codon Platform
- Strategies with a shear sensitive clone
- Glycan profile optimization to reduce afucosylated species
- ADCC optimization
- Growing NS0 cells in disposable bioreactors
- Multi- factors evaluation to improve charge variances via DoE
- Steady state perfusion with high cell density and viability
- FTE-Model Approach
- Protein Refolding Analysis using HT-Approach
- Trouble-shooting product issues in GMP manufacturing
Case study: Developability Assessment
Background
Client asked WuXi Biologics to assess the key attributes of two candidate molecules to determine the lead molecule for further product CMC development.
Challenge
Design developability platform, identify key selection criteria and key product attributes to determine the lead clone and molecule.
Solution/Results
- Both candidate molecules had the same product pool titer.
- Molecule 1 exhibited aggregation and purification issues and lower solubility.
- Taking all factors into consideration (see examples in Table 1), Molecule 2 was selected for cell line cloning and CMC development.
- After further development and optimization, a titer of 9.7 g/L was achieved and the IND-filing was submitted in April 2016.
Table 1 – Results
| Criteria | Molecule 1 | Molecule 2 |
|---|---|---|
| Fit to Upstream | 3.7 g/L | 3.7 g/L |
|
Fit to Downstream |
>10% HMW level after ProA 52% CEX step yield |
87% CEX step yield |
|
Formulation and Stability |
Platform Solubility: 100mg/ml | Platform Solubility: >150mg/ml |
|
Biophysical and Biochemical Characterization |
DSC: Tm2: 76.4oC | DSC: Tm2: 78.7oC |
Case study: Improving the Product Quality with WuXia AI Codon
Background
Client requested a cell line with good titer and product quality.
Challenge
With the third party codon, high titer clones tend to have more aggregates.
Solution
-
Used
AI codon program for codon optimization
-
Evaluated multiple sequences with Transient expression.
- Selected one sequence with higher titer and better product quality for stable cell line development.
Results
- Delivered clones with titer of 4 g/L and SEC main peak above 98%.
| Criteria | Result |
|---|---|
| Titer |
Pool Titer: 1.5 g/L Clone Titer: 3.6 g/L |
|
Monoclonality |
Documented pre-plating ViCell pictures and ClonePix colony pictures |
AI Based Codon Optimization
Case study: Strategies with a shear sensitive clone
Background
- The initial process had a low growth, high lactate and very low titer of 0.2 g/L.
Challenge
- The clone was shear sensitive and grew poorly with microsparger.
Solution/Results
- The WuXi Biologics team replaced microsparger with drilled-hole sparger & optimized bioreactor conditions.
- We further improved the process by basal & fed-batch media/feed optimization.
- The effort led to a much better cell growth, cell viability, lactate profile and an improved titer of >1.5 g/L.
Case study: Glycan profile optimization to reduce afucosylated species
Background
- The initial process had high afucosylated species with toxicity concern
Challenge
- Afucosylated species >15%, needs to control at below 10%
- Titer impact needs to be manageable while making process changes.
Solution/Results
- Applying a new pH control strategy can successfully decrease afucosylation level.
- No titer compromise under the new pH control strategy.
- Improved process was successfully scaled up to 500L GMP manufacture.
Case study: ADCC optimization
Background
- The initial process had significantly lower ADCC activity when compared to the reference material.
Challenge
- Dramatically lower ADCC activity (about 50% lower), which could be hard to optimize
- CDC activity needs to stay within range while optimizing ADCC activity
Solution/Results
- Adding medium components allowed ADCC activity adjustment and without significant impact on CDC activity. Both ADCC and CDC were within target range after optimization.
- Strong correlation between ADCC activity and total a-fucosylated glycan content (G0+G1+G2+Man5).
- Valuable tool for molecules that need ADCC functions.
Case study: Growing NS0 cells in disposable bioreactors
Background
- Direct transfer of process from client to WuXi Biologics for clinical trial production.
- Client required comparability assessment to the legacy manufacturing process and final product.
Challenge
- No developer of the original process was available. Had to perform only document-based process transfer.
- New process to be conducted in single-use bioreactors while the original process was performed through fixed stainless steel tanks/pipeline.
- Cell culture of NS0 cell line is difficult in single-use disposable systems according to historical records.
Solution/Results
- Through extensive PD effort, the manufacturing process was successfully transferred, optimized and performed in the single-use systems.
- Process scale-up in single-use systems was confirmed at 50 L and 250 L scale and likewise GMP manufacturing was conducted at 2,000 L scale in single-use bioreactors.
- Comparability data in IND filing has been accepted by the U.S. FDA.
- The 1st biological drug manufactured in China for global clinical trials was successfully produced by WuXi Biologics’ manufacturing site in Wuxi city.
Case study: Multi- factors evaluation to improve charge variances via DoE
Background
- The process had drastic change in charge variant profile after media change in an effort to reduce CoGs during late phase development. The goal is to optimize the process to match acidic peak species in the original process.
Challenge
- The acidic peak must be reduced by ~10%.
Solution/Results
- A DoE study was executed in spin tube scale down model, where 7-factors were employed.
- The optimized process was confirmed in 3L bioreactors with 10 % acidic peak reduction.
Case Study: Steady state perfusion with high cell density and viability
Background
- Recombinant protein with high demand in productivity and product quality
Challenge
- Relatively instable product
- PQA very sensitive to cell culture performance such as viability
Solution/Results
- Specific productivity is limited due to the nature of the protein, high viable cell density is required to achieve high productivity
- ATF system was introduced as cell retention device to achieve high viable cell density; proper cell removal (bleeding) rate was used to maintain stable viable cell density and viability
- Long term steady state perfusion (typically 60 days, up to 130 days) with stable cell growth performance, metabolism, productivity and PQA
- Clarified harvest from ATF enables direct product capture downstream process
- Perfusion process successfully scaled up to 250L (with 150L working volume) GMP runs
Case Study: FTE-Model Approach
Background
WuXi Biologics suggested using an FTE approach to meet critical bispecific antibody development timelines.
Challenge
Previous client attempts to produce and purify the protein led to undesirable expression levels of a bispecific antibody due to low expression of one of the antibody chains which resulted in only 10% of desired protein product from the entire protein production campaign. These challenges were delaying product CMC development and thus time was critical as original IND filing date was in jeopardy.
Solution/Results
After discussion with the client and outlining the DOE study required to potentially solve the problem, client-dedicated personnel via an FTE program was established. The client-dedicated FTE team generated and evaluated 42 different constructs and evaluated expression levels of the bispecific antibody in over 1,000 minipools using a high-throughput methodology. Minipools demonstrating best results were moved to clone screening and eventual final clone selection. Results provided in Table 3.
Table 3 – FTE program results
| Final Titer of bispecific Ab clone | Main product expression levels before purification | Main product purity after purification | Date of client IND filing |
|---|---|---|---|
| 3.8 g/L | 90% | 98% | (On-time filing) |
Case Study: Protein Refolding Analysis using HT-Approach
Background
Client struggled with developing the tools to understand if a recombinant cytokine product produced in e. coli had refolded correctly after removal from inclusion bodies post fermentation.
Challenge
Develop a high-throughput system that could quickly evaluate over 130 different molecules using multiple analytical methods to meet critical client timelines and budget.
Solution/Results
A well-established, HT matrix-screening method in a fast, efficient way.
Case Study: Trouble-shooting product issues in GMP manufacturing
Background
Client could not get a consistent product quality or GMP manufacturing process performance at current vendor. Requested WuXi Biologics assemble a team from multiple functional areas to trouble-shoot manufacturing process and provide solutions.
Challenge
Product had high HMW and dimer content and low enzymatic activity. Had to evaluate over 800 pages of batch records, perform technical transfer and trouble-shoot based on investigation master plan in short time frame.
Solution/Results
You are leaving WuXi Biologics Website, after which our Privacy Notice will not apply. Please keep it in mind the protection of your privacy. Are you willing to proceed?