# US 10,308,928 B2 · System for generating high concentration factors for low cell density suspensions > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 10,308,928 B2 - Document ID: US10308928B2 - Status: Granted - Kind: B2 - Application number: 15/690,190 - Filed: 2017-08-29 - Published: 2018-01-18 - Granted: 2019-06-04 - Earliest claimed: 2013-09-13 - Assignees: FloDesign Sonics, Inc. - Applicants: FloDesign Sonics, Inc. - Inventors: Bart Lipkens; Jason Dionne; Goutam Ghoshal - Web page: https://patentgrep.com/patent/US10308928B2 ## CPC - C12N 13/00: Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves - B01D 17/044: {by changing the pressure} - B01D 21/28: Mechanical auxiliary equipment for acceleration of sedimentation, e.g. by vibrators or the like - B01D 21/283: {Settling tanks provided with vibrators} - B06B 1/0644: {using a single piezoelectric element (B06B1/0688 takes precedence)} - C12M 47/02: {Separating microorganisms from the culture medium; Concentration of biomass (separating microorganisms from their culture media C12N1/02)} - G10K 15/00: Acoustics not otherwise provided for - G10K 15/043: {producing shock waves (G10K15/046, G10K15/06 take precedence; generating seismic energy G01V1/02)} - H10N 30/2023: {having polygonal or rectangular shape} - H10N 30/8554: {Lead-zirconium titanate [PZT] based} - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - 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) - 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) - 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} - G10K: SOUND-PRODUCING DEVICES; METHODS OR DEVICES FOR PROTECTING AGAINST, OR FOR DAMPING, NOISE OR OTHER ACOUSTIC WAVES IN GENERAL; ACOUSTICS NOT OTHERWISE PROVIDED FOR - H10N: ELECTRIC SOLID-STATE DEVICES NOT OTHERWISE PROVIDED FOR ## Abstract Acoustophoretic devices and methods for concentrating targeted biological cells in a reduced volume using multi-dimensional acoustic standing waves are disclosed. The methods include flowing a mixture of a host fluid and the biological cells through an acoustophoretic device. The acoustophoretic devices include an inlet, an outlet, and a flow chamber having an ultrasonic transducer-reflector pair. Biological cells, such as T cells, are separated from a host fluid for utilization in allergenic or autologous cell therapies. The disclosed devices and methods are capable of concentrating biological cells to at least 100 times their original cell concentration. The disclosed methods and devices are further capable of decreasing an original feed volume to a final concentrated volume that is less than one percent of the original feed volume. ## Claims ### Claim 1 A method for concentrating target particles in a host fluid, comprising: receiving an original concentration mixture of the host fluid and the target particles in an acoustophoretic device, the acoustophoretic device comprising: an acoustic chamber with an inlet; at least one ultrasonic transducer coupled to the acoustic chamber, the at least one ultrasonic transducer configured to launch an acoustic wave in the acoustic chamber; and a collector with at least one angled wall that taper downwards in a cross-sectional area located between the inlet and the at least one ultrasonic transducer; exciting the at least one ultrasonic transducer to launch the acoustic wave in the acoustic chamber; and concentrating the target particles via the acoustic wave by at least an order of magnitude difference from the original concentration mixture. ### Claim 2 (depends on 1) The method of claim 1, further comprising concentrating the target particles to a final concentration of about 150 times to about 300 times the original concentration mixture. ### Claim 3 (depends on 1) The method of claim 1, further comprising concentrating the target particles to a final concentration of about 300 times to about 600 times the original concentration mixture. ### Claim 4 (depends on 1) The method of claim 1, wherein the target particles are cells with an original cell concentration of about 1 million cells per mL. ### Claim 5 (depends on 1) The method of claim 1, further comprising recovering the target particles in a final concentrated volume from the acoustic chamber. ### Claim 6 (depends on 5) The method of claim 5, wherein the total particle retention in the final concentrated volume is at least 80%. ### Claim 7 (depends on 1) The method of claim 1, further comprising flowing the original concentration mixture vertically into the acoustic chamber. ### Claim 8 (depends on 1) The method of claim 1, further comprising: flowing the original concentration mixture into an inlet of the acoustophoretic device; recovering a particle depleted permeate at a first outlet; and recovering the concentrated target particles at a second outlet. ### Claim 9 (depends on 8) The method of claim 8, further comprising collecting the concentrated target particles with the collector. ### Claim 10 (depends on 9) The method of claim 9, further comprising flowing the original concentration mixture through an annular plenum around the collector. ### Claim 11 (depends on 9) The method of claim 9, further comprising continuously trapping the target particles via the acoustic wave, such that the target particles agglomerate, aggregate, clump, or coalesce together, and settle out of the host fluid and into the collector. ### Claim 12 (depends on 1) The method of claim 1, wherein the target particles are one or more of T-cells, B cells, or NK cells. ### Claim 13 (depends on 1) The method of claim 1, further comprising reducing a turbidity of the host fluid by at least 65% after 60 minutes, or at least 70% after 60 minutes, or at least 80% after 60 minutes, or at least 90% after 60 minutes. ### Claim 14 A method for obtaining concentrated target cells, comprising: receiving an original feed volume of a mixture of a host fluid and the target cells in an acoustophoretic device, the acoustophoretic device comprising: an acoustic chamber with an inlet; at least one ultrasonic transducer coupled to the acoustic chamber, the at least one ultrasonic transducer configured to launch an acoustic wave in the acoustic chamber; and a collector with at least one angled wall that tapers downwards in a cross-sectional area located between the inlet and the at least one ultrasonic transducer; exciting the at least one ultrasonic transducer to launch the acoustic wave in the acoustic chamber; and recovering at least 40% of the concentrated target cells in the original feed volume in a final concentrated volume via the acoustic wave, wherein the final concentrated volume is at least an order of magnitude smaller than the original feed volume. ### Claim 15 (depends on 14) The method of claim 14, wherein the total target cell retention in the final concentrated volume is at least 80%. ### Claim 16 (depends on 14) The method of claim 14, further comprising continuously trapping the target cells via the acoustic wave, such that the target cells agglomerate, aggregate, clump or coalesce together, and settle out of the host fluid. ### Claim 17 (depends on 14) The method of claim 14, further comprising reducing a turbidity of the host fluid by at least 65% after 60 minutes, or at least 70% after 60 minutes, or at least 80% after 60 minutes, or at least 90% after 60 minutes. ### Claim 18 A method for obtaining concentrated target cells, comprising: receiving an original feed volume of a mixture of a host fluid and the target cells in an acoustophoretic device, the acoustophoretic device comprising: an acoustic chamber with an inlet; at least one ultrasonic transducer coupled to the acoustic chamber, the at least one ultrasonic transducer configured to launch an acoustic wave in the acoustic chamber; and a collector with at least one angled wall that taper downwards in a cross-sectional area located between the inlet and the at least one ultrasonic transducer; exciting the at least one ultrasonic transducer to generate the acoustic wave in the acoustic chamber; concentrating the target cells via the acoustic standing wave; and retaining at least 80% of the target cells from the original feed volume in a final concentrated volume.