End-to-end process support, from development to industrialization
Adeno-associated viral (AAV) vectors are the most commonly used delivery system for gene therapies currently under development. AAV comes in a wide range of serotypes, which allows for targeting different types of tissue specifically. Other advantages include that it is non-pathogenic and cannot replicate on its own, which increases safety; and its ability to transduce non-dividing and dividing cells.
Much progress has been made in developing commercial production-scale processes for gene therapies in recent decades, but challenges persist, especially around upscaling. We offer a wide range of robust, scalable solutions and have the technical knowledge and experience to help you every step of the way.
Most AAV vector production follows the same basic process, with some variation as needed.
Adherent Cell Seeding
Early on in therapeutic development, viral vectors are usually manufactured in adherent cells using two-dimensional systems such as cell factories or roller bottles. As processes scale up and more virus is needed, the footprint and manual operations required to grow cells in this fashion quickly becomes problematic.
The Xpansion Multiplate Bioreactor is a closed, single-use system that has been specifically designed for the scale-up of shear-sensitive, adherent mammalian cells under controlled conditions. It provides a similar microenvironment to a cell factory but in a much more compact space and with fully automated environmental control.
Adherent Cell Culture
As the therapies progress into clinical development, there is a requirement to increase quantity and quality of the virus produced. Manufacturing virus from adherent cells using fixed-bed bioreactor technology allows for direct transfer of the 2D reference process, which minimizes risk and saves time.
The iCELLis bioreactor is a fully-integrated, high-cell density bioreactor designed to simplify processes involving adherent cells by combining the advantages of single-use technologies with the benefits of a bioreactors - reduced handling, footprint, labor, cost, and improved process assurance, robustness, and scalability. The iCELLis technology can be used at bench-scale (0.5 to 4 m2) and manufacturing-scale (66 to 500 m2).
Suspension Cell Seeding & Cell Culture
Manufacturers are increasingly looking to produce viral vectors for gene therapy in suspension cell cultures, as the pressure increases for increased scalability. The cell seed train for suspension cultures generally occurs in smaller bioreactors, which may also double as small batch production vessels. As production increases, larger volume bioreactors are needed. Viral vector production in suspension culture usually requires additional testing and optimization. Our experienced team of applications experts can work with you to determine the best solution for your upstream processes.
Allegro™ STR Bioreactors
Allegro STR single-use stirred tank bioreactors were thoughtfully designed to maximize usability and process assurance while delivering consistent and scalable cell culture performance across a full range of maximum working volumes, from 50 – 2000 L.
Allegro™ XRS Bioreactors
The Allegro XRS 25 bioreactor system is a single-use bioreactor system with unique agitation and control properties designed for the cultivation of mammalian cells in suspension culture under controlled conditions at the 2 to 25 L scale.
The first process step after cell culture is the removal of cells, cell debris, and other impurities to reduce biological burden as much as possible. The easiest and most economical technology to clarify the cell culture is filtration. Low yield is a common challenge in this step, especially as cell densities increase upstream and sizes of viral particle increase. Our experienced technical team can help you achieve high throughput and yield by optimizing protocols and parameters for this step, in addition to selecting the best filter materials and size.
Stax™ Disposable Depth Filter Systems
This ready-to-use platform is scalable in size and performance to meet lab, pilot and process needs. Stax disposable capsules are available in three interchangeable capsule sizes ranging from 0.25 m2 to 2.0 m2 (2.70 ft2 to 21.5 ft2). For commercial production scale, large Stax capsules with single-layer V100P media grade or dual-layer PDK11 offer high throughput and recoverability for manufacturing of viral vector products.
Supracap™ 100 Depth Filter Capsules
Supracap 100 capsules integrate seamlessly into any development process with its versatile configurations, sizes and available filter media. They offer easy scalability, and the fully disposable design eliminates any cleaning or handling concerns usually associated with housings, especially where biological and hazardous products are to be filtered. Supracap 100 with single-layer V100P media grade capsules offer high throughput and recoverability for viral vector products.
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Reducing bioburden through a suitable grade 0.2 µm or 0.45 µm filter is usually the next step in this process. The type of filter used here can vary based on the titer and volume of material, fouling properties of the fluid, and processing time requirements.
Bioburden Reduction Filters
Our technologically innovative bioburden control filters reduce bacterial loads in process feeds to acceptably low levels for successful and cost-effective protection of processes from microbial and particulate contamination. Membranes are inert and have low protein and excipient binding properties suitable for all types of drug products. Available in a wide range of formats: cartridges for stainless steel installations and capsules for single-use systems.
All process development-scale filter formats are designed to scale directly to larger cGMP filter configurations for fast and easy production scale-up.
Ultrafiltration/Diafiltration using Tangential Flow Filtration (TFF) membranes further concentrates the target molecule and exchanges buffers. Achieving high yield is a common challenge with TFF, but our technical experts can work with you to optimize your UF/DF unit operation based on experience and thorough testing.
Single-Use Tangential Flow Filtration (TFF) Modules
Single-use tangential flow filtration (SUTFF) modules are ready-to-use gamma-irradiated cassette modules that can be integrated into single-use TFF set-ups. Integrating the process-proven Omega™ polyethersulfone (PES) membrane, single-use TFF modules are available in scalable formats from 93 cm² up to 2.5 m² of membrane area.
T-Series TFF Cassettes with Omega Membrane
T-Series cassettes with Omega membrane are ideal for development, pilot, and production-scale TFF applications in diverse biological and biopharmaceutical processes. They are especially useful in vaccine and conjugate concentration and diafiltration, purification and recovery of monoclonal antibodies (mAb) or recombinant proteins, and blood plasma fractionation and purification.
T-Series TFF Cassette Holders
TFF cassette holders for the medium and large-scale production environment have a small footprint, yet still are capable of holding large areas of membrane. All stainless steel holders are engineered to precise specifications with a high standard internal and external surface finish and are available with either manual torque (MT) or auto torque (AT) compression systems for ease of use.
The polishing step removes empty capsids, DNA and host cell proteins (HCP). Adsorptive membrane technology offers an efficient and effective option for polishing gene therapy products.
Mustang® Q Membrane Chromatography Capsules
We have combined our competencies in membrane device design and chromatography to develop a range of high-performance, scalable Mustang Q XT membrane chromatography capsules. These capsules are reliable, easy to use and enable users to significantly improve their process economics through reduced buffer consumption, increased throughput and reduced capital expenditure.
Mustang Q XT Acrodisc
Disposable membrane chromatography units for scale-down process development. Mustang XT Acrodisc units, available in Q and S chemistries, are ideal for scale-down process development work in a variety of downstream process applications. A membrane volume of < 1 mL reduces the amount of sample required for evaluation, and the female luer lock inlet and outlet simplifies connection to typical low pressure chromatography systems.
Bulk Drug Substance Filtration
Manufacturing for clinical purposes requires the production of highly pure and biologically active vectors that meet regulatory requirements. A last sterilizing grade (0.2 µm) filtration step is usually needed before product is sent for final filling.
Sterile Liquid Filtration
Optimized sterilizing-grade liquid filtration in your drug manufacturing process drives efficiencies while providing confidence in achieving drug critical quality attributes. Selecting the right sterile filter media and format, for your specific process helps to maximize yield and recovery of valuable drug product and ensure production efficiency. Our extensive range of sterilizing-grade liquid filters and formats enable us to offer filtration solutions for all applications and drug types, from mAbs, recombinant proteins, vaccines and gene therapies to small molecule drugs, and plasma derivatives.
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Pall has a global team of over 300 experienced technical professionals to support your molecule from pre-clinical to commercial manufacturing. From process development services and end-to-end platforms, to on-site testing and applications support for specific process steps, Pall will help you find the best solution based on data and testing.