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Published on: August 16, 2019
Development of an ELP-Z based mAb affinity precipitation process using scaled-down filtration techniques
Rahul D Sheth1, Bharat V Bhut2, Mi Jin2
1Department of Chemical and Biological Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY 12180, United States; Biologics Process Development, Bristol-Myers Squibb, East Syracuse, NY, United States.
A novel elastin-like polypeptide-Z domain fusion (ELP-Z) based precipitation process for monoclonal antibody (mAb) purification is developed. This method utilizes scaled-down filtration techniques, demonstrating efficient recovery and impurity clearance, with an alternative filtration strategy for improved scalability.
Area of Science:
- Biotechnology
- Protein Engineering
- Bioprocess Engineering
Background:
- Monoclonal antibody (mAb) purification is critical in biopharmaceutical manufacturing.
- Current purification methods can be complex and costly.
- Affinity precipitation offers a potential alternative for efficient mAb recovery.
Purpose of the Study:
- To develop and evaluate an elastin-like polypeptide-Z domain fusion (ELP-Z) based affinity precipitation process for mAb purification.
- To assess the scalability of the ELP-Z precipitation process using membrane filtration techniques.
- To optimize filtration strategies for efficient precipitate recovery and impurity removal.
Main Methods:
- Development of an ELP-Z fusion protein for mAb affinity precipitation.
- Application of Tangential Flow Filtration (TFF) for precipitate recovery and diafiltration.
- Analysis of precipitate morphology and formation kinetics using confocal microscopy.
- Implementation of normal flow filtration (NFF) as an alternative recovery strategy.
Main Results:
- TFF enabled complete recovery of ELP-Z-mAb and ELP-Z precipitates with high impurity clearance.
- ELP-Z precipitates caused significant membrane fouling during TFF, leading to flux decay.
- Confocal microscopy revealed differences in precipitate morphology and formation kinetics impacting TFF performance.
- An optimized NFF strategy achieved high volumetric throughput and 96% ELP-Z recovery during mAb elution.
Conclusions:
- The ELP-Z affinity precipitation process is a viable method for mAb purification.
- Scaled-down filtration techniques, particularly NFF, can be effectively employed for process scale-up.
- Understanding precipitate characteristics is crucial for optimizing filtration performance in affinity precipitation processes.
