End-to-end cell therapy bioprocessing device for continuous-flow enrichment, washing, and electrotransfection of target cells
A system for cell bioprocessing and cell therapy manufacturing can include a series of microfluidic modules to enable continuous-flow end-to-end cell bioprocessing. Each module can implement a different technology, and the modules can be coupled to one another to perform various unit operations in the cell bioprocessing or cell-therapy manufacturing chain to enable direct processing of a blood or blood product sample. The system can automatically and continuously process the sample into genetically-modified lymphocytes or T cells for cellular therapy. The technologies implemented by each module in the system can include any combination of microfluidic acoustophoresis, microfluidic acoustophoretic media exchange or cell washing, and continuous-flow microfluidic electrotransfection. Modules implementing these microfluidic technologies can be interconnected with plastic tubing or with a custom manifold.
1 . A system, comprising:
an inlet channel that receives a target fluid flow comprising target particles;
an acoustophoresis device that receives the target fluid flow comprising the target particles from the inlet channel and moves the target particles from the target fluid flow to a buffer fluid flow comprising cargo particles;
an electroporation device downstream from the acoustophoresis device that receives the buffer fluid flow comprising the target particles and the cargo particles, and applies an electric field to the buffer fluid flow to cause a portion of the target particles in the buffer fluid flow to absorb a portion of the cargo particles;
a conductivity sensor coupled to an inlet of the electroporation device and configured to measure a conductivity of the buffer fluid flow received by the electroporation device and provide a signal indicating the conductivity;
an outlet channel that provides an output buffer fluid flow from the electroporation device; and
a controller configured to receive the signal from the conductivity sensor and control a pump to adjust a flow rate of the target fluid flow into the inlet channel based on the conductivity.
2 . The system of claim 1 , wherein the acoustophoresis device comprises:
a central channel that receives the buffer fluid flow comprising the cargo particles from a source of buffer fluid and the target fluid flow from the inlet channel; and
a piezoelectric transducer coupled to the central channel that causes the target particles to move from the target fluid flow to the buffer fluid flow in the central channel.
3 . The system of claim 1 , wherein the electroporation device comprises:
a central channel receiving the buffer fluid flow from the acoustophoresis device and a conductive buffer flow from a second channel; and
an electrode electrically coupled to a portion of the central channel that provides the electric field.
4 . The system of claim 1 , wherein the target particles in the target fluid flow are lymphocytes, and the target fluid flow further comprises waste particles including at least one of red blood cells, granulocytes, or monocytes.
5 . The system of claim 1 , wherein the inlet channel is a first inlet channel, and further comprising:
a second inlet channel that receives an input fluid flow comprising waste particles and the target particles; and
an acoustic separation device that receives the input fluid flow comprising the target particles and the waste particles from the second inlet channel and separates the input fluid flow into the target fluid flow comprising the target particles and a waste fluid flow comprising the waste particles.
6 . The system of claim 5 , wherein the waste fluid flow is transported via one or more waste channels to a waste reservoir, and the target fluid flow that exits from the acoustic separation device is transported via a second outlet channel into a target reservoir; and
wherein the system further comprises a pump that transports the target fluid flow from the target reservoir to the acoustophoresis device via the second inlet channel.
7 . The system of claim 1 , further comprising:
the pump, wherein the pump transports the target fluid flow from an input reservoir to the acoustophoresis device via the inlet channel; and
another pump that transports the buffer fluid flow comprising the target particles and the cargo particles from the acoustophoresis device to the electroporation device via an intermediate channel.
8 . The system of claim 1 , further comprising one or more holding reservoirs between two or more of the inlet channel, the acoustophoresis device, the electroporation device, or the outlet channel.
9 . The system of claim 1 , further comprising:
a separation device that receives the output buffer fluid flow from the outlet channel and separates enriched target particles in the output buffer fluid flow from waste particles in the output buffer fluid flow; and
an output reservoir that receives the enriched target particles from the separation device.
10 . The system of claim 1 , wherein connections between two or more of the inlet channel, the acoustophoresis device, the electroporation device, or the outlet channel comprise at least one of poly-vinyl-chloride tubing or silicone tubing.
11 . The system of claim 10 , further comprising:
one or more fluid capacitors coupled with at least one of the connections, the one or more fluid capacitors configured to regulate a flow rate of at least one fluid in the system.
12 . The system of claim 1 , further comprising:
one or more sensors configured to transmit, to the controller, signals representing a density value of waste particles or the target particles in at least one of the target fluid flow, the buffer fluid flow, or the output buffer fluid flow.
13 . The system of claim 1 , further comprising:
one or more flow sensors that transmit, to the controller, one or more signals representing a flow rate of a fluid flowing through at least one of the inlet channel, the acoustophoresis device, the electroporation device, or the outlet channel.