small molecules
biologics
cell & gene
bioscience
capsules
Exosomes are small membrane vesicles that are secreted by a multitude of cell types, which play a role in cell to cell communication. They can serve as clinically valuable tools for early diagnosis, prognosis and potentially targeted treatment.
From a manufacturing stand-point, the challenge resides not only in the production but also in the characterization. Exosomes have the potential to make cell and gene therapies commercially viable for large populations.
We are still at a very early stage and most developers of these innovative technologies are in pre-clinical or early clinical stages, working to demonstrate the efficacy and the potential of exosomes-based therapies.
Prime biological cargo delivery
Exosomes are produced by all cells. They are a sub-group of extracellular vesicle with a size range typically between 30 and 150 nm, have a Phospholipid Bilayer shell with specific surface markers, and a genetic package containing of RNA, DNA and proteins.
Some cells can use exosomes to transfer genetic material from one to another. Specifically, an exosome can transfer microRNAs to control the expression of a gene, and messenger RNAs to manufacture proteins. They also transfer proteins themselves, lipids, DNA and others.
Exosomes are acting as cargo delivery systems in our body to deliver specific packages with positive or negative messages from one cell to another in a targeted way at distance. Simply put, they are a fresh and well conserved source of biomarkers coming from live cells which reflect those of the parenteral cells.
So what stands in the way?
Aside from decoding the biology and mechanism for therapeutic effect in this novel field, these are some key challenges faced by exosomes-based therapy developers. Those are linked to the very early and pre-clinical stage which most of the exosomes-based therapies are in. There are limited characterization methods available, a need for precise isolation and purification methods and few partners who can provide access to end-to-end GMP manufacturing expertise. Identifying optimal formulation, obtaining starting cell-lines, media, gene editing & cargo loading can be a challenge, as well as defining functional assays.
No mountain high enough
Choosing the right partner becomes critical in such a pioneering field. By leveraging our knowledge in 2D/3D cell culture and our viral manufacturing expertise, alongside the latest exosome characterization technologies, we can provide a complete path for exosome manufacturing.
Upstream
Fully controlled and scalable (up/out) exosome generation technologies
Downstream and fill & finish
Selective exosome isolation technologies
Characterization
A complete set of measurement tools to enable both upstream and downstream process development as well as quality control and product characterization
Characterization
| ZetaView® (NTA) for particle sizing and concetration determination | FACS, qPCR and ELISA for analyzing exosomes and extracted exosome cargo | Further characterization techniques currently on trial in R&D, targeted mid-2020 for GMP use |
- NTA - nanoparticle tracking analysis
- FACS - FLourescence-activated cell sorting
- qPCR - Real-time quantitative polymerase-chain-reaction
- ELISA - enzyme-linked immunosorbent assay
Upstream
| Source material | Cell line engineering | Expansion |
|---|---|---|
| Tissue acquisition and cell isolation (or exising cell line) | Nucleofection / electroporation: Lonza Nucleofector™ and other electroporation methods
Lentiviral and AAV transduction methods |
Large-scale 2D systems: CellSTACK®s and cell factories
Large-scale 3D systems: stir tank bioreactor (adherent and aggregate based cultures), Wave Bioreactor™ (suspension based cultures), hollow fiber (Quantum®) |
Downstream
| Isolation / purification | Formulation implementation at DS | Delivery |
|---|---|---|
| Tangential flow filtration (TFF) for clarification and volume reduction
Size-exclusion (SEC) / Ion-exchange for selection of exosomes Ultracentrifugation |
Compounding of buffer and target excipient composition
Process parameters to achieve final composition of exosome in bulk DS |
Shipping and storage
End-to-end traceability, identification and injection |
Formulation and Fill & Finish Drug Product
| Formulation development | DP process development and design | GMP DP manufacturing and QC testing |
|---|---|---|
| Selection of appropriate buffer and excipients to have optimal formulatuon for improved stability
Development services for lyophilization of exosome for 5°C storage Freeze / thaw and agitation study to support handling and transportation |
Stability studies, manufacturing compatibility, particle assessment & ID testing*, light stability Overfills, container closure system selection & qualification, container closure integrity in frozen state* Extractables / leachables testing, in-use clinical administration design (pharmacy manual) and testing |
GMP DP manufacturing
QC release and stability studies (including project specific development) and implementation of Pharmacopeia methods Implementation of purity methods from customer for QC release and stability |