Acoustic separation of particles for bioprocessing
Inventors
Fiering, Jason O. • Kotz, Kenneth T.
Assignees
Charles Stark Draper Laboratory Inc
Publication Number
US-12332235-B2
Publication Date
2025-06-17
Expiration Date
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Abstract
A method for separating particles in a biofluid includes pretreating the biofluid by introducing an additive, flowing the pretreated biofluid through a microfluidic separation channel, and applying acoustic energy to the microfluidic separation channel. A system for microfluidic separation, capable of separating target particles from non-target particles in a biofluid includes at least one microfluidic separation channel, a source of biofluid, a source of additive, and at least one acoustic transducer coupled to the microfluidic separation channel. A kit for microfluidic particle separation includes a microfluidic separation channel connected to an acoustic transducer, a source of an additive, and instructions for use.
Core Innovation
The invention provides methods and systems for separating target particles from non-target particles in a biofluid by pretreating the biofluid, flowing the pretreated biofluid into an inlet of a microfluidic separation channel, and applying acoustic energy to the microfluidic separation channel to accumulate target particles within a primary stream and accumulate non-target particles within a secondary stream along the separation channel. The invention also provides a system comprising at least one microfluidic separation channel, a source of biofluid, a source of additive, and at least one acoustic transducer coupled to a wall of the microfluidic separation channel, and a kit comprising a microfluidic separation channel connected to an acoustic transducer, a source of an additive, and instructions for use. The methods and systems are described for selective, differential separation of synthetic target particles (including carrier, capture, enrichment, and delivery particles) from mixed suspensions of cells and particles in a biofluid.
The background identifies that foreign particles introduced into a biofluid for therapeutic treatment may pose a safety risk if injected into a patient and that former washing of biofluid samples has been performed by batch or continuous centrifugations, magnetic separation, or membrane filtration. The patent explains limitations of these prior approaches, including centrifugation's limited ability to separate particles of similar density and membrane filtration's general reliance on size exclusion which can fail when foreign capture particles are similar in size to desired cells. The disclosure frames a need for improved methods to remove introduced particles from a liquid suspension of mixed cell types prior to therapeutic delivery.
The disclosure teaches improving selective acoustic separation by pretreating the biofluid with an additive to alter one or more physical properties of the biofluid, target particles, or non-target particles (for example density, size, compressibility, or aggregation potential), flowing the pretreated biofluid through microfluidic separation channels, and applying acoustic energy (for example a standing acoustic wave transverse to flow) to drive differential accumulation into primary and secondary streams. The systems and methods are described as capable of continuous operation, providing separation by multiple physical parameters (for example size and density), being amenable to scaling, and achieving a high degree of purification with physiologically acceptable additives. The disclosure further describes system features such as sensors, control modules, manifolds, recycle lines, and post-treatment to enable regulated pretreatment and acoustic separation in-line or in staged arrangements.
Claims Coverage
The patent includes eleven main inventive features extracted from two independent system claims.
Microfluidic separation channel with dual outlets and collection channels
At least one microfluidic separation channel comprising at least one inlet, a first outlet, and a second outlet, and a first collection channel in fluid communication with the first outlet and a second collection channel in fluid communication with the second outlet.
Source of biofluid in fluid communication with separation channel
A source of the biofluid in fluid communication with the at least one inlet of the at least one microfluidic separation channel.
Source of additive configured to alter physical properties
A source of an additive in fluid communication with the source of the biofluid, configured to introduce at least one additive into the biofluid, the additive capable of altering one or more physical properties of at least one of the target particles, non-target particles, or biofluid.
Acoustic transducer coupled to separation channel wall
At least one acoustic transducer coupled to a wall of the at least one microfluidic separation channel.
Recycle line for target particle depleted fluid
A recycle line extending from the second collection channel in fluid communication with the source of the biofluid, configured to recycle target particle depleted fluid from the second outlet to the source of the biofluid.
Sensor measuring HCT or particle concentration
At least one sensor configured to measure at least one of hematocrit (HCT %) of the biofluid and concentration of target particles or non-target particles.
Control module introducing predetermined additive volume in response to sensor
A control module in electrical communication with the at least one sensor and the source of the additive, configured to introduce a predetermined volume of the additive into the biofluid in response to a measurement of at least one of HCT % of the biofluid and concentration of the target particles or the non-target particles in the biofluid.
Standing acoustic wave transverse to channel to drive particles to node or anti-node
An acoustic transducer positioned to apply a standing acoustic wave transverse to the microfluidic separation channel to drive the particles to a node or anti-node within the separation channel.
Output sensor and control to regulate output HCT
At least one output sensor configured to measure at least one parameter of an output suspension (for example HCT % or concentration of target or non-target particles) and a control module in electrical communication with the output sensor and the acoustic transducer, configured to regulate at least one input parameter of the acoustic transducer or to regulate additive volume to control the HCT % of the output suspension.
Control module configured to regulate HCT of output to less than about 1%
The additive is configured to regulate the HCT % of the output suspension, and the control module is configured to regulate the HCT % of the output suspension to less than about 1% by regulating a volume of the additive introduced to the microfluidic separation channel.
Thermal management for acoustic transducer
A heat sink configured to dissipate heat generated by the acoustic transducer (for example a thermoelectric cooler) to prevent warming of fluids flowing through the separation channel.
The independent claims disclose a microfluidic acoustic separation system that integrates an additive source and automated sensing/control (including feedback to introduce a predetermined additive volume) with acoustic transduction, recycle capability, output sensing, and thermal management to effect regulated separation of target and non-target particles in a biofluid.
Stated Advantages
Removal of particles can be performed continuously.
Separation by both size and density to further enhance particle separation.
Separation processes may be readily scaled to small or large sample volumes.
A high degree of purification can be achieved with the addition of safely injectable, physiologically acceptable additives.
Improved selective separation of target particles from a biofluid suspension by introducing an additive to alter physical properties of the fluid, target particles, or non-target particles.
Documented Applications
Processing of cells for cell therapy and bioprocessing, including removal of synthetic cell capture particles from blood samples.
Large scale bioprocessing such as separation of carrier particles from cell culture samples and harvesting cultured cells (for example mesenchymal stem cells on carrier particles).
Separation of magnetic or paramagnetic capture particles from final therapeutic products.
Diagnostic assays, environmental monitoring assays, tissue engineering, and in vitro models.
Biomanufacturing systems, including examples for energy applications.
In-line collection, separation, post-treatment, and delivery of biofluid between donor and recipient subjects.
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