Method using a three-dimensional bioprocessor
Inventors
Ling, Jian • Zimmern, Kreg A. • Milone, Michael C.
Assignees
University of Pennsylvania Penn • Southwest Research Institute SwRI
Publication Number
US-11492580-B2
Publication Date
2022-11-08
Expiration Date
2040-05-12
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Abstract
Described herein is a beads-free bioprocessor as an automated and cost-effective T cell processing and manufacturing platform. T cells are a core component in CAR T cell therapies for cancer treatment, but are difficult to manufacture to scale in clinically relevant quantities. The 3D bioprocessor provides an alternative device that is scalable, beads-free, easy-to-use, and cost-effective for using CAR T cell therapy in cancer immunotherapy. Besides CAR T cell application, this platform technology has potential for many other applications such as cancer cell isolation.
Core Innovation
The invention introduces a beads-free, three-dimensional (3D) bioprocessor platform designed for automated and cost-effective T cell processing and manufacturing. The 3D bioprocessor comprises a plurality of spheres interconnected by rods, forming a fixed-bed structure with a high surface-to-volume ratio. This structure facilitates the binding of cells, such as T cells, by enabling the immobilization of biotinylated antibodies on its surface, which in turn allows for specific cell activation and expansion.
This innovation addresses significant limitations associated with current CAR T cell manufacturing methods that rely on magnetic microbeads. Traditional processes require labor-intensive manual steps, are open systems susceptible to contamination, and are not cost-effective. Furthermore, these methods present challenges in separating T cells from beads, introduce risks of non-T cell contamination, and often demand specialized manufacturing facilities to maintain sterility. The described 3D bioprocessor eliminates the dependency on beads and offers a scalable, easy-to-use, and single-use alternative suitable for both autologous and allogenic cell therapy applications.
Central to the method is supplying the 3D bioprocessor with adjustable structural parameters, coating its surface with a protein (such as avidin or streptavidin), and immobilizing one or more biotinylated antibodies (for example, anti-CD3, anti-CD28, or anti-CD2). Cells are then flowed through this structure, where specific cell types bind to their cognate antibody, enabling controlled activation, expansion, and even isolation or purification within a closed and automated system.
Claims Coverage
There are two independent claims in this patent, each covering a distinctive aspect of the 3D bioprocessor method and apparatus for cell processing.
Beads-free 3D bioprocessor structure for cell binding
The invention covers a method for cell processing that utilizes a 3D bioprocessor comprising: - A plurality of spheres and a plurality of rods interconnecting the spheres, where rods provide spacing between adjacent spheres. - Application of a coating on the surface area (including the spheres and rods). - Attachment of a protein (such as avidin or streptavidin) to the coating. - Immobilization of one or more biotinylated antibodies onto this protein. - Flowing cells through the 3D bioprocessor so that cells bind to the one or more biotinylated antibodies.
Repeatable mesh apparatus with dimensional and compositional specification
The method also is defined for a bioprocessor that: - Comprises a plurality of spherical beads interconnected by rods, with dimensions specified (e.g., spherical beads of about 1 mm to about 3 mm, rods less than half the sphere diameter, rods cylindrical, organized in one or more layers, with each layer possibly offset). - Forms a repeatable and non-random mesh structure with substantial uniformity. - Has a coating and protein with immobilized biotinylated antibodies on its surface. - Ensures that binding of cells to the antibodies occurs as cells are flowed through the structure.
In summary, the inventive features claim both the beads-free 3D sphere-rod bioprocessor method for cell binding, activation, and expansion, and the specific apparatus structure with its material, dimensional, and operational properties for efficient and scalable cell processing.
Stated Advantages
The bioprocessor offers a low cost, easy-to-manufacture, easy-to-use, and single-use platform for cell processing in cell and gene therapy.
Provides a high surface-to-volume ratio fixed-bed with adjustable structural parameters and scalability.
Eliminates reliance on magnetic beads, avoiding cellular uptake and supply limitations.
Forms a repeatable and non-random mesh structure, ensuring consistency of bioprocess and hydrodynamic forces on cells.
Allows for a closed, automated system that reduces manual labor and contamination risk, supporting compliance with GMP requirements.
Enables controlled shear stress and perfusion flow, maximizing T-cell binding and activation.
Easily scalable by 3D printing for different clinical applications, including both autologous and allogenic therapies.
Documented Applications
Manufacturing platform for CAR T cell therapies in cancer immunotherapy.
Cancer cell isolation and purification using antibody-coated surfaces for specific binding.
Cell separation, isolation, and purification from mixed cell populations such as immunological cells (T cells, NK cells, B cells).
Activation and expansion of T cells and other cell types for cell and gene therapy.
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