# US 9,752,114 B2 · Bioreactor using acoustic standing waves

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- Number: US 9,752,114 B2
- Document ID: US9752114B2
- Status: Granted
- Kind: B2
- Application number: 15/453,582
- Filed: 2017-03-08
- Published: 2017-06-22
- Granted: 2017-09-05
- Earliest claimed: 2012-03-15
- Assignees: FloDesign Sonics, Inc.
- Applicants: FloDesign Sonics, Inc.
- Inventors: Bart Lipkens; Louis Masi; Stanley Kowalski, III; Walter M. Presz, Jr.; Jason Dionne; Brian Dutra; Ari Mercado; Thomas J. Kennedy, III; Arthur Martin; John Rozembersky
- Web page: https://patentgrep.com/patent/US9752114B2

## CPC

- C12M 47/10: {Separation or concentration of fermentation products (bioreactors combined with means for distillation or extraction of liquid fuel C12M43/02)}
- B01D 17/04: Breaking emulsions
- B01D 17/06: Separation of liquids from each other by electricity
- B01D 21/283: {Settling tanks provided with vibrators}
- B01J 19/10: employing sonic or ultrasonic vibrations
- B01J 19/18: Stationary reactors having moving elements inside (B01J19/08, B01J19/26 take precedence)
- B06B 1/0644: {using a single piezoelectric element (B06B1/0688 takes precedence)}
- C12M 23/14: {Bags}
- C12M 23/22: {Transparent or translucent parts (glassware for laboratory use B01L3/00)}
- C12M 27/02: {Stirrer or mobile mixing elements}
- C12M 29/10: {Perfusion}
- C12M 29/18: {External loop; Means for reintroduction of fermented biomass or liquid percolate (loop type reactors for chemical or physical processes B01J19/2435)}
- C12M 33/08: {by vibration}
- C12M 33/14: {with filters, sieves or membranes}
- C12M 35/04: {Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli}
- C12M 47/02: {Separating microorganisms from the culture medium; Concentration of biomass (separating microorganisms from their culture media C12N1/02)}
- B01J 2219/00094: Jackets
- C12M: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; {APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS}
- B01D: SEPARATION (separating solids from solids by wet methods B03B, B03D; by pneumatic jigs or tables B03B; by other dry methods B07; magnetic or electrostatic separation of solid materials from solid materials or fluids, separation by high-voltage electric fields B03C; centrifuges, vortex apparatus B04; presses per se for squeezing-out liquid from liquid-containing material B30B9/02; treatment of water C02F, e.g. softening by ion-exchange C02F1/42; {arrangements of air intake cleaners in gas turbine plants F02C7/05} ; arrangements or mounting of filters in air-conditioning, air-humidification or ventilation F24F13/28)
- B01J: CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B06B: METHODS OR APPARATUS FOR GENERATING OR TRANSMITTING MECHANICAL VIBRATIONS OF INFRASONIC, SONIC, OR ULTRASONIC FREQUENCY, {e.g.} FOR PERFORMING MECHANICAL WORK IN GENERAL (for particular applications, see the relevant subclasses, e.g. B07B1/40, B23Q17/12, B24B31/06; measurement of mechanical vibrations G01H; in direction finding, locating, distance or velocity measuring G01S; {generating seismic energy G01V1/02}; control of mechanical vibrations in general G05D; sound-producing devices, e.g. bells, sirens, whistles G10K, {e.g. methods or devices for transmitting, conducting, or directing sound in general G10K11/00}; generation of electrical oscillations H03B; electromechanical resonators in general H03H; electromechanical transducers {for communication techniques, e.g. microphones, speakers} H04R)

## Abstract

A perfusion bioreactor includes at least one ultrasonic transducer that can acoustically generate a multi-dimensional standing wave. The standing wave can be used to retain cells in the bioreactor, and can also be utilized to dewater or further harvest product from the waste materials produced in a bioreactor.

## Claims

### Claim 1

A system comprising:
a bioreactor;
a primary clarification stage, the primary clarification stage including an acoustophoretic separator downstream of and fluidly connected to the bioreactor, the acoustophoretic separator comprising:
a flow chamber; and
an ultrasonic transducer and a reflector located opposite the ultrasonic transducer and configured to produce a multi-dimensional standing wave in the flow chamber when the ultrasonic transducer is actuated; and
a secondary clarification stage downstream of the primary clarification stage.

### Claim 2 (depends on 1)

The system of claim 1, wherein the secondary clarification stage includes a filtration device fluidly connected to the acoustophoretic separator.

### Claim 3 (depends on 2)

The system of claim 2, wherein the filtration device is one or more of a depth filter, a sterile filter, a tangential flow filter, or a crossflow filter.

### Claim 4 (depends on 1)

The system of claim 1, wherein the secondary clarification stage is configured to isolate a desired product by size exclusion filtration.

### Claim 5 (depends on 1)

The system of claim 1, wherein the secondary clarification stage includes a plurality of filtration devices arranged in series.

### Claim 6 (depends on 5)

The system of claim 5 wherein at least one of the plurality of filtration devices is a separation column and another of the plurality of filtration devices is a porous filter, the porous filter being located upstream of the separation column and fluidly connected thereto.

### Claim 7 (depends on 1)

The system of claim 1, wherein the multi-dimensional standing wave includes an axial force component and a lateral force component which are of the same order of magnitude.

### Claim 8 (depends on 1)

The system of claim 1, wherein the secondary clarification stage is adapted to trap at least some material that passes through the primary clarification stage.

### Claim 9 (depends on 1)

The system of claim 1, wherein the secondary clarification stage includes a second acoustophoretic separator comprising a second ultrasonic transducer and opposed reflector configured to produce a second multi-dimensional standing wave in the second acoustophoretic separator when the second ultrasonic transducer is actuated, the second multi-dimensional standing wave being adapted to trap at least some material that passes through the primary clarification stage.

### Claim 10 (depends on 1)

The system of claim 1, wherein the bioreactor is a perfusion bioreactor.

### Claim 11

A process of isolating and recovering desired product from a cell culture media, comprising:
receiving the cell culture media at a primary clarification stage, the primary clarification stage including an acoustophoretic separator downstream of and fluidly connected to the bioreactor, wherein the acoustophoretic separator comprises:
a flow chamber; and
an ultrasonic transducer and a reflector located opposite the ultrasonic transducer;
providing an output of the acoustophoretic separator to a secondary clarification stage downstream of the primary filtration stage; and
recovering the desired product from the secondary clarification stage.

### Claim 12 (depends on 11)

The process of claim 11, further comprising sending the cell culture media from the flow chamber back to the bioreactor through a recycle outlet of the acoustophoretic separator.

### Claim 13 (depends on 11)

The process of claim 11, wherein the secondary clarification stage includes a filtration device fluidly connected to the acoustophoretic separator.

### Claim 14 (depends on 13)

The process of claim 13, wherein the filtration device is one or more of a depth filter, a sterile filter or a crossflow filter.

### Claim 15 (depends on 11)

The process of claim 11, further comprising isolating the desired product by size exclusion filtration.

### Claim 16 (depends on 11)

The process of claim 11, further comprising arranging the secondary clarification stage as a plurality of filtration devices arranged in series.

### Claim 17 (depends on 16)

The process of claim 16, wherein at least one of the plurality of filtration devices is a separation column and another of the plurality of filtration devices is a porous filter, the porous filter being located upstream of the separation column and fluidly connected thereto.

### Claim 18 (depends on 11)

The process of claim 11, further comprising actuating the ultrasonic transducer to generate a multi-dimensional standing wave with an axial force component and a lateral force component which are of the same order of magnitude.

### Claim 19 (depends on 11)

The process of claim 11, further comprising trapping at least some material with the secondary clarification stage that passes through the primary clarification stage.

### Claim 20 (depends on 11)

The process of claim 11, further comprising actuating an ultrasonic transducer in a second acoustophoretic separator in the secondary clarification stage to produce a multi-dimensional standing wave in the second acoustophoretic separator, the multi-dimensional standing wave being adapted to trap at least some material that passes through the primary clarification stage.