Acoustic perfusion devices

Patent metadata

Field	Value
Publication number	US-9752113-B2
Application number	US-201615139187-A
Country	US
Kind code	B2
Filing date	Apr 26, 2016
Priority date	Mar 15, 2012
Publication date	Sep 5, 2017
Grant date	Sep 5, 2017

Abstract

Acoustic perfusion devices for separating biological cells from other material in a fluid mixture are disclosed. The devices include an inlet port, an outlet port, and a collection port that are connected to an acoustic chamber. An ultrasonic transducer creates an acoustic standing wave in the acoustic chamber that permits a continuous flow of fluid to be recovered through the collection port while keeping the biological cells within the acoustic chamber to be returned to the bioreactor from which the fluid mixture is being drawn.

First claim

The invention claimed is:

1. An acoustic perfusion device, comprising:
an acoustic chamber;
an inlet port and an inlet flow path leading from the inlet port to the acoustic chamber;
an outlet port and an outlet flow path leading from the acoustic chamber to the outlet port for recirculating a fluid mixture;
at least one collection port for collecting harvest fluid from the fluid mixture; and
at least one ultrasonic transducer and at least one reflector opposite the at least one ultrasonic transducer, wherein the at least one ultrasonic transducer includes a piezoelectric material configured to be driven by a voltage signal to create a first multi-dimensional acoustic standing wave across a collection flow path leading from the acoustic chamber to the at least one collection port; and
a secondary flow chamber configured to permit the harvest fluid to pass through a second acoustic standing wave with a frequency different from or equal to the first multi-dimensional acoustic standing wave to further clarify the harvest fluid.

2. The device of claim 1 , wherein the acoustic chamber is below the at least one collection port.

3. The device of claim 1 , wherein the inlet flow path is shaped to generate a tangential flow path below an acoustic field generated by the acoustic standing wave.

4. The device of claim 1 , wherein a pressure rise and an acoustic radiation force on cells are generated at an upstream interface region of the acoustic standing wave to clarify fluid passing through the acoustic standing wave.

5. The device of claim 1 , wherein the at least one reflector is made of a transparent material.

6. The device of claim 1 , wherein the outlet port is below the inlet port.

7. The device of claim 1 , having a total of two or more ultrasonic transducers located on multiple sides of the collection flow path.

8. The device of claim 7 , where the at least one reflector is located between the two or more ultrasonic transducers.

9. The device of claim 7 , wherein the two or more ultrasonic transducers are within the collection flow path and the at least one reflector is opposite the two or more ultrasonic transducers in a wall of the device.

10. The device of claim 1 , wherein the at least one transducer is an array.

11. The device of claim 1 , wherein the first multi-dimensional acoustic standing wave results in an acoustic radiation force having an axial force component and a lateral force component that are of the same order of magnitude.

12. The device of claim 1 , having two or more collection ports spaced apart from each other on the top end of the device.

13. The device of claim 1 , wherein both a planar acoustic standing wave and the first multi-dimensional acoustic standing wave are created across the collection flow path.

14. The device of claim 1 , further comprising a recirculation flow path between the inlet port and the outlet port that does not enter the acoustic chamber, wherein the recirculation flow path is located below the acoustic chamber.

15. The device of claim 14 , wherein the inlet flow path travels through a different passage than the outlet flow path.

16. The device of claim 14 , wherein the inlet flow path and the outlet flow path travel through a common passage.

17. The device of claim 1 , wherein the fluid mixture is made up of a primary fluid and particles.

18. The device of claim 17 , where the particles comprise mammalian cells, bacteria, cell debris, fines, proteins, exosomes, vesicles, viruses, or insect cells.

19. A method for separating biological cells from a fluid mixture, comprising:
flowing the fluid mixture containing the biological cells through an acoustic perfusion device, the device comprising:
an acoustic chamber;
an inlet port, an inlet flow path leading from the inlet port to the acoustic chamber;
an outlet port for recirculating the fluid mixture and the biological cells;
at least one collection port for collecting harvest fluid; and
at least one ultrasonic transducer below the at least one collection port and at least one reflector opposite the at least one ultrasonic transducer, wherein the at least one ultrasonic transducer includes a piezoelectric material driven by a voltage signal to create a first multi-dimensional acoustic standing wave across a collection flow path leading from the acoustic chamber to the at least one collection port; and
a secondary flow chamber in which the harvest fluid passes through a second acoustic standing wave with a frequency different from or equal to the first multi-dimensional acoustic standing wave to further clarify the harvest fluid;

driving the at least one ultrasonic transducer to create the acoustic standing wave; and
collecting a fluid mixture enriched in cells from the outlet port and collecting a harvest fluid mixture depleted in cells from the at least one collection port.

20. The method of claim 19 , wherein the outlet port is below the inlet port.

21. The method of claim 19 , wherein a flow rate through the collection flow path is at least one order of magnitude smaller than a flow rate through the inlet flow path.

22. The method of claim 19 , wherein the inlet flow path leads from the inlet port downwards towards the outlet port and then upwards to the acoustic chamber, creating a recirculating fluid stream that is locally substantially tangential to the at least one acoustic standing wave.

23. The method of claim 22 , wherein the recirculating fluid stream transports away cells that are constantly held back at an interface region of the acoustic standing wave.

24. The method of claim 19 , wherein a pressure rise and an acoustic radiation force on cells are generated at an upstream interface region of the acoustic standing wave to clarify fluid passing through the acoustic standing wave.

25. The method of claim 19 , wherein a flow rate of the fluid mixture entering the device through the inlet port is about 1 liter per minute and a flow rate of the harvest fluid depleted in cells exiting the device through the at least one collection port is about 10 milliliters per minute.

26. The method of claim 19 , further comprising passing the harvest fluid through the secondary flow chamber to further clarify the harvest fluid.

27. A flow device, comprising:
at least one inlet for receiving a flowing mixture of a primary fluid and cells,
a first ultrasonic transducer configured to produce a first multi-dimensional acoustic standing wave and a pressure rise and an acoustic radiation force on cells, generated at an upstream interface region of the first multi-dimensional acoustic standing wave, to separate the flowing mixture into a primary high cell concentration fluid stream and a secondary harvest fluid stream;
an outlet port for the primary high cell concentration fluid stream;
a bleed port for extracting a concentrated fluid/cell mixture; and
at least one collection port for the secondary harvest fluid stream; and
a secondary flow chamber containing a second ultrasonic transducer that produces a second acoustic standing wave with a frequency different from or equal to the first multi-dimensional acoustic standing wave to further clarify the secondary harvest fluid stream.

28. The device of claim 27 , where the secondary flow chamber is configured to further clarify the secondary harvest fluid stream by trapping and subsequently clumping, cluster

Assignees

• Flodesign Sonics Inc

Inventors

• Lipkens Bart
• Miller Erik
• Ross-Johnsrud Benjamin
• Presz Jr Walter M
• Chitale Kedar
• Kennedy Iii Thomas J
• Winiarski Lauren

Classifications

• C12M29/18

  External loop; Means for reintroduction of fermented biomass or liquid percolate (loop type reactors for chemical or physical processes B01J19/2435) · CPC title

• C12N13/00

  Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves · CPC title

• B01D21/283

  Settling tanks provided with vibrators · CPC title

• C12M33/08

  by vibration · CPC title

• B06B1/0644

  using a single piezoelectric element (B06B1/0688 takes precedence) · CPC title