Acoustic separation of T cells
Acoustophoretic devices and methods for separating biological cells (particularly T-cells) from other fluids/materials using multi-dimensional acoustic standing waves are disclosed. The devices include an inlet, at least two outlets, and a flow chamber having an ultrasonic transducer-reflector pair. Specifically, T cells, B cells, or NK cells can be separated from other blood components. A dual-pass acoustophoretic system including two acoustophoretic devices arranged in series and fluidly connected to one another is also illustrated. Means for pre-chilling the mixture prior to separation in the devices or system can be used to improve retention, concentration, and clarification and to prevent outgassing.
1. A method for separating biological cells from a host fluid, comprising:
chilling a mixture of the host fluid and the biological cells;
flowing the cooled mixture of the host fluid and the biological cells through a first acoustophoretic device, the first acoustophoretic device comprising:
a flow chamber including at least one inlet and at least one outlet;
at least one ultrasonic transducer coupled to the flow chamber, the at least one ultrasonic transducer including a piezoelectric material configured to be driven to create a multi-dimensional acoustic standing wave in the flow chamber; and
a reflector opposite to the at least one ultrasonic transducer; and
driving the at least one ultrasonic transducer to create the multi-dimensional standing wave in the flow chamber, such that at least some of the biological cells are trapped in the standing wave, and agglomerate, aggregate, clump, or coalesce together, and settle out of the host fluid due to enhanced gravitational settling forces.
2. The method of claim 1 , wherein the biological cells are Jurkat T cells, B cells, or NK cells.
3. The method of claim 1 , wherein the biological cells are T cells having a low ribosomal content of less than 30 wt %.
4. The method of claim 1 , wherein the mixture is chilled to a temperature of about 20° C. to about 25° C.
5. The method of claim 1 , wherein the chilling lowers the temperature of the mixture by from about 10° C. to about 20° C.
6. The method of claim 1 , wherein the first acoustophoretic device further comprises at least one concentrate outlet located at a bottom end of the flow chamber for recovering the biological cells; and the first acoustophoretic device also further comprises a permeate outlet located at a top end of the flow chamber.
7. The method of claim 6 , further comprising sending the host fluid and biological cells exiting the permeate outlet of the first acoustophoretic device to an inlet of a second acoustophoretic device.
8. The method of claim 6 , wherein fluid recovered from the at least one concentrate outlet (i) has a cell concentration of at least two times an original cell concentration of the mixture of the host fluid and the biological cells; and (ii) has a volume of at least one half an original feed volume of the mixture of the host fluid and the biological cells.
9. The method of claim 1 , wherein the mixture is flowed into the first acoustophoretic device at a flow rate such that the mixture has a residence time of at least 5 minutes.
10. An acoustophoretic system for separating biological cells from a mixture of a host fluid and the biological cells, comprising:
a chiller for chilling the mixture; and
a first acoustophoretic device comprising:
a flow chamber including at least one inlet and at least one outlet, the at least one inlet being fluidly connected to the chiller; and
at least one ultrasonic transducer coupled to the flow chamber, the at least one ultrasonic transducer including a piezoelectric material configured to be driven to create a multi-dimensional acoustic standing wave in the flow chamber.
11. The system of claim 10 , further comprising a reflector opposite to the at least one ultrasonic transducer.
12. The system of claim 10 , wherein the chiller for chilling the mixture is a pre-chiller or a fan.
13. The system of claim 10 , wherein the at least one inlet of the first acoustophoretic device is a dump diffuser.
14. The system of claim 10 , wherein the first acoustophoretic device further comprises at least one concentrate outlet located at a first end of the flow chamber, and wherein the first acoustophoretic device also further comprises a permeate outlet located at a second end of the flow chamber opposite the first end.
15. The system of claim 14 , further comprising a second acoustophoretic device comprising:
a flow chamber including at least one inlet and at least one outlet, the at least one inlet being fluidly connected to the permeate outlet of the first acoustophoretic device;
at least one ultrasonic transducer coupled to the flow chamber, the at least one ultrasonic transducer including a piezoelectric material configured to be driven to create a multi-dimensional acoustic standing wave in the flow chamber; and
a reflector opposite to the at least one ultrasonic transducer.
16. A device for separating biological cells from a host fluid, comprising:
means for chilling a mixture of the host fluid and the biological cells;
means for flowing the cooled mixture of the host fluid and the biological cells through a first acoustophoretic device;
means for separating the biological cells from the host fluid using a multi-dimensional acoustic standing wave.
17. The device according to claim 16 , further comprising means for collecting the separated biological cells.
18. The device according to claim 16 , further comprising means for collecting a permeate.