WO2023065019A1

WO2023065019A1 - Sonicated bioreaction system and method

Info

Publication number: WO2023065019A1
Application number: PCT/CA2022/051529
Authority: WO
Inventors: Alexander Down; Ryan YAU
Original assignee: Alexander Down; Yau Ryan
Priority date: 2021-10-20
Filing date: 2022-10-17
Publication date: 2023-04-27

Abstract

A bioreaction system and related methods are disclosed. The system comprises a sonic array comprising two or more transducers, the sonic array being configured to selectively emit sonication, each of the two or more transducers being configured to be in direct or indirect contact with the cell culture, the sonication having sonication parameters; and a controller operably coupled to and in communication with the sonic array for selectively adjusting the sonication parameters and for receiving signals from the two or more transducers.

Description

SONICATED BIOREACTION SYSTEM AND METHOD

Cross-Reference to Related

[0001] This application claims the benefit of United States Provisional Application No.

63/257,880, filed October 20, 2021, and United States Provisional Application No. 63/282,116, filed November 22, 2021, the contents of which applications are hereby incorporated by reference in their entireties.

Field

[0002] The present disclosure generally relates to bioreaction, and more specifically to a sonicated bioreaction system and related methods.

[0003] Liquid-suspended plant cell cultures are routinely used to produce metabolites for pharmaceutical products, food additives, or energy. The yield of these cultures is critical to the economy of the process. Research performed in small-scale laboratory settings has demonstrated a correlation between sonic radiation (also referred to as "sonication") on a specific cell culture and an improvement in one of the following cell metrics of that specific cell culture: cell growth (which refers to cell growth rate and total growth (biomass) accumulated in a growth cycle); anti-clumping (also referred to as "deflocculation"); metabolite production (also referred to as "elicitation"); metabolite release from the cell without negatively impacting cell viability (also referred to as "perfusion culture"); and cell destruction (e.g., for metabolite recovery in cases of batch culturing where the biomass is discarded after metabolite extraction). [0004] The present disclosure aims to provide a bioreaction system and related methods for sonicating a cell culture to improve one or more cell metrics thereof on a large, industrial scale.

[0005] According to a broad aspect of the present disclosure, there is provided a bioreaction system for use on a cell culture, the bioreaction system comprising: a sonic array comprising two or more transducers, the sonic array being configured to selectively emit sonication, each of the two or more transducers being configured to be in direct or indirect contact with the cell culture, the sonication having sonication parameters; and a controller operably coupled to and in communication with the sonic array for selectively adjusting the sonication parameters and for receiving signals from the two or more transducers.

[0006] In some embodiments, the controller is configured to determine or generate, based on the signals, one or more of: a density of the cell culture; an attenuation of the cell culture; a particle size distribution of the cell culture; and a multi-dimensional image of the cell culture.

[0007] In some embodiments, the two or more transducers are spaced apart from one another.

[0008] In some embodiments, a plurality of the two or more transducers are configured to form a beam steering array.

[0009] In some embodiments, each of the two or more transducers is one or more of: a piezoelectric transducer, an electromagnetic transducer, and a magnetostrictive transducer.

[0010] In some embodiments, the controller comprises one or more of: a processer, a memory, a display, and a user interface. [0011] In some embodiments, at least one of the two or more transducers is configured to perform flow-through sonication.

[0012] In some embodiments, the bioreaction system comprises a tank for receiving the cell culture therein.

[0013] In some embodiments, the tank has a volume in a range from about 500L to about 5000L.

[0014] In some embodiments, a wall of the tank is made of a disposable membrane.

[0015] According to another broad aspect of the present disclosure, there is provided a method comprising: taking ultrasonic measurements of a cell culture by two or more transducers of a sonic array, the two or more transducers being in direct or indirect contact with the cell culture; based on the ultrasonic measurements, performing one or more of: determining a particle size distribution of the cell culture; determining a relative attenuation of the cell culture; determining a relative density of the cell culture; and generating a multi-dimensional image of the cell culture; and correlating one or more of the particle size distribution, the relative attenuation, the relative density, and the multi-dimensional image to a growth stage of the cell culture.

[0016] In some embodiments, the method comprises, based on the growth stage, determining whether the cell culture is at a peak of metabolite concentration.

[0017] In some embodiments, the method comprises, upon determining that the cell culture is at the peak of metabolite concentration, generating a signal to an operator.

[0018] In some embodiments, the method comprises, based on one or more of the particle size distribution, the relative attenuation, the relative density, and the multi-dimensional image, providing an indicator for one or more of: a yield of the cell culture; a yield per culture time of the cell culture; a harvest suitability of the cell culture, and a metabolite concentration of the cell culture.

[0019] In some embodiments, the method comprises emitting sonication, by at least one of the two or more transducers, to the cell culture, the sonication having sonication parameters. [0020] In some embodiments, the method comprises, based on the growth stage, adjusting at least one of the sonication parameters.

[0021] In some embodiments, the method comprises determining whether the growth stage has a corresponding programmed profile, the programmed profile having profile parameters; and upon determining that the growth stage has the corresponding programmed profile, causing the sonication parameters to match the profile parameters.

[0022] In some embodiments, the method comprises pasteurizing the cell culture by the at least one of the two or more transducers.

[0023] In some embodiments, the pasteurizing is performed in-situ or flow-through.

[0024] In some embodiments, the method comprises performing metabolite extraction on the cell culture by the at least one of the two or more transducers.

[0025] In some embodiments, the metabolite extraction is performed in-situ or flow-through.

[0026] The details of one or more embodiments are set forth in the description below. Other features and advantages will be apparent from the specification and the claims. Brief of the

[0027] The invention will now be described by way of an exemplary embodiment with reference to the accompanying simplified, diagrammatic, not-to-scale drawings. Any dimensions provided in the drawings are provided only for illustrative purposes, and do not limit the invention as defined by the claims. In the drawings:

[0028] FIG. 1 is a graphical representation of a metabolite concentration curve of a cell culture during a culturing process, according to one embodiment.

[0029] FIG. 2 is a schematic view of a bioreaction system, according to one embodiment of the present disclosure.

[0030] FIG. 3 is a flowchart illustrating a process that can be carried out using the bioreaction system, according to one embodiment of the present disclosure.

Detailed Description of the Embodiments

[0031] When describing the present invention, all terms not defined herein have their common art-recognized meanings. To the extent that the following description is of a specific embodiment or a particular use of the invention, it is intended to be illustrative only, and not limiting of the claimed invention. The following description is intended to cover all alternatives, modifications and equivalents that are included in the scope of the invention, as defined in the appended claims.

[0032] Based on the existing literature on sonication, one may conclude that for every culture species, the improvement of each cell metric of that culture species requires a different sonication parameter. Further, the improvement of each metric may be optimized during a specific growth stage of that culture species.

[0033] The bioreaction system and methods described herein aim to improve more than one cell metric for a single cell culture on a large, or may be even industrial, scale. In some embodiments, the bioreaction system and methods take ultrasonic measurements of the cell culture fluid and monitor the growth stage or metabolite concentration of the cell culture based on the ultrasonic measurements. While it is not an explicit measurement of growth stage or metabolite concentration, the ultrasonic measurement can be correlated to the growth stage or metabolite concentration of the cell culture. The bioreaction system and methods herein can thus be used to monitor growth stage or metabolite concentration without the need to take physical samples of the cell culture or the need for the conventional costly and labour-intensive methods of growth measurement and metabolomic analysis. The ultrasonic measurements may be used to determine the optimal window for harvesting the cell culture. In some embodiments, the bioreaction system can modulate the ultrasonic emission accordingly for a culture growth stage to improve a specific cell metric.

[0034] In some embodiments, ultrasonic measurements of two or more batches of the same cell culture can be taken continuously or frequently, and optionally simultaneously to, for example, (i) generate a growth curve or metabolite concentration curve for that cell culture; and/or (ii) identify batches that deviate from the norm, which may be an indicator of upstream processing issues. By taking real-time ultrasonic measurements, the bioreaction system and methods herein may allow continued iterations of any process variables and corresponding results to be recorded, observed, and/or analyzed. The bioreaction system and methods can be used to collect real-time data on many batches of cell cultures, sequentially or simultaneously, with little or no human operator intervention. In some embodiments, the bioreaction system can be retrofitted into existing cell culturing plant equipment. Accordingly, the bioreaction system and methods herein may be cost-effectively deployed for monitoring metabolite concentrations in real-time and for optimizing the cell culturing process on a large scale.

[0035] During the stationary growth phase of a culture species, the concentration of metabolites can be a non-linear function of time. A sample metabolite concentration curve 12 for a cell culture during the culturing process is shown in FIG. 1. For any cell species, the metabolite concentration may peak (/.e., reach a higher level than previous levels) more than once over a period of time. For example, curve 12 has a peak A, occurring at about day 14, where the metabolite concentration is at the absolute maximum over the observed time span of 14 days. Curve 12 also has a peak B, occurring at about day 9, where the metabolite concentration is less than that at peak A, but is greater than those of the previous days. In the sample embodiment, peak B provides the highest yield per culture time.

[0036] In some embodiments, depending on the upstream and/or downstream processes intended for the cell culture, it may be advantageous to harvest the cell culture at peak A (e.g., where it is desirable to achieve maximum metabolite concentration) or peak B (e.g., where time is of the essence). Conventionally, metabolomic analysis involving either mass spectrometry or nuclear magnetic resonance spectrometry is used to determine the concentration of metabolites for a culture species. Mass spectrometry and nuclear magnetic resonance spectrometry both require manual sample preparation by skilled technicians and costly equipment, and generally cannot provide metabolite concentration data for culture species in real-time. [0037] After the culturing process, one of the downstream processing steps may be pasteurization. In some embodiments, the cell culture may be pasteurized in-situ where the culturing process took place. In other embodiments, the bioreaction system may perform continuous flow ("flow-through") pasteurization on the cell culture if higher ultrasonic intensities are required for the target microbial reduction. Another post-culturing downstream processing step may be metabolite extraction. In some embodiments, the bioreaction system and methods herein allow some in-situ metabolite extraction by sonication, which may yield higher extraction percentages than prior art methods. In some embodiments, the bioreactions system and methods may perform flow-through metabolite extraction by sonication on the cell culture. Deflocculation is another possible post-culturing downstream processing step.

[0038] FIG. 2 shows one embodiment of a bioreaction system 20 for sonicating cell cultures. Cell cultures may include, for example, plant cell cultures, animal cell cultures, fungal cell cultures, protist cell cultures, bacterial cell cultures, a combination of any of the foregoing cultures, and the like. In some embodiments, system 20 may be used for suspension cultures. In some embodiments, the system 20 comprises a tank 22 for accommodating therein a suspended cell culture. In other embodiments, tank 22 is not part of the system 20 but rather system 20 is configured to engage with the tank 22. The suspended cell culture in FIG. 2 is denoted by "CC". The tank 22 has an inlet (not shown) for receiving the cell culture CC into the tank and an outlet 28 for extracting the cell culture CC from the tank. In some embodiments, tank 22 has an internal volume of about 500L to about 5000L. In some embodiments, at least a portion of the wall of the tank 22 is a disposable membrane. In some embodiments, the tank 22 is a fillable bag made of a disposable membrane. [0039] Bioreaction system 20 comprises a sonic array 24 comprising a plurality of transducers 24a, 24b, 24c, 24d. The sonic array 24 is operably coupled to the tank 22 for determining various parameters (e.g., the growth stage) of the cell culture CC. Each of the transducers 24a, 24b, 24c, 24d may be piezoelectric, electromagnetic, and/or magnetostrictive. In some embodiments, the transducers 24a, 24b, 24c, 24d operate to emit sonication to the cell culture CC and may be selectively programmable to emit sonication having specific parameters. The parameters of sonication include, for example: frequency, power, duty cycle, and pulse timing. The transducers 24a, 24b, 24c, 24d also operate to take ultrasonic measurements of the cell culture CC in the tank at the respective locations of the transducers. In some embodiments, the transducers 24a, 24b, 24c, 24d are configured to take ultrasonic measurements in real-time.

[0040] In some embodiments, the sonic array 24 has a transducer 24a that is positioned at the bottom of the tank 22 and penetrates the tank 22 to directly contact the cell culture CC at the bottom of the tank 22. In some embodiments, the sonic array 24 has a transducer 24b that is installed inside the tank 22 so that transducer 24b is in direct contact with the top of the cell culture CC. In some embodiments, the sonic array 24 has one or more transducers 24c, 24d attached to the wall at the outer surface of the tank 22 for indirect ("through-wall") contact with the cell culture CC.

[0041] Where at least a portion of the tank 22 is made of a disposable membrane, the bioreaction system 20 may be configured such that the transducer 24c, 24d comes into physical contact with the outer surface of the tank 22 when the tank 22 is at least partially filled with the cell culture CC. For example, where the tank 22 is a fillable bag, the transducers 24c, 24d may be secured to a structure, such a frame or cage, configured to receive the bag 22 therein. When the bag 22 is at least partially filled with the cell culture, hydrostatic pressure inside the bag 22 forces the wall of the bag 22 radially outwardly to abut against and be in physical contact with the transducers 24c, 24d. In other embodiments, a coupling material may be applied to the interface between the wall of the tank 22 and the transducers 24c, 24d.

[0042] In some embodiments, the sonic array 24 has a plurality of transducers, positioned at any or all of the aforementioned positions relative to the tank 22. Using multiple transducers in the sonic array 24 may increase sonication power transfer to the cell culture CC and/or improve the resolution or sensitivity of the resulting ultrasonic measurements. Where the sonic array 24 has a plurality of transducers 24a, 24b, 24c, 24d, the transducers are spaced apart from one another. For example, where the sonic array comprises two transducers 24c, 24d attached to the outer wall of the tank 22, the transducers 24c, 24d are spaced apart axially, laterally, and/or circumferentially on the tank 22 by a distance. In another example, where the sonic array 24 comprises a bottom transducer 24a and an internal transducer 24b, the transducers 24a, 24b are spaced apart from one another by a distance.

[0043] In some embodiments, the system 20 comprises a controller 26 operably coupled to and in communication with the sonic array 24. The controller 26 is configured to periodically determine one or more metrics of the cell culture CC based on signals generated by the transducers 24a, 24b, 24c, 24d of the sonic array 24. In some embodiments, the sonication output of one or more of the transducers 24a, 24b, 24c, 24d can be controlled by controller 26 such that controller 26 can selectively adjust the parameters of the sonication emitted by the transducers. In some embodiments, the controller 26 comprises a processor, a memory for storing data, and optionally a user interface and/or a display. [0044] In some embodiments, based on the transducers' signals, the controller 26 continuously or periodically determines a spatial averaged density of the cell culture CC in real-time using time- of-flight transmission technique. In other embodiments, the controller 26 continuously or periodically determines a spatial averaged attenuation in real-time using amplitude transmission modality. These measurements of spatial averaged density and spatial averaged attenuation require at least two transducers 24a, 24b, 24c, 24d to be positioned in or on the tank 22 so that the sonication emitted by one transducer can be received by the other transducer(s).

[0045] In some embodiments, based on the transducers' signals, the controller 26 continuously or periodically generates a real-time multi-dimensional image of density or attenuation of the cell culture CC. Generating the multi-dimensional image may require that some of the transducers of the sonic array 24 form a beam steering array, which may allow more accurate growth stage determination of the cell culture CC in the tank 22, for example, where the cell culture is heterogeneous. The techniques for forming a beam steering array and the corresponding digital controls for operating same are known to those skilled in the art.

[0046] In some embodiments, based on the transducers' signals, the controller 26 continuously or periodically determines the particle size distribution of the cell culture CC in the tank 22.

[0047] In some embodiments, the measurements and/or results generated by the controller 26 based on the transducers' signals can be correlated to a growth stage of the cell culture CC in the tank 22 so that the growth stage at any given time can be determined by the controller 26. In some embodiments, the controller 26 may determine the growth stage of the cell culture CC by multi-variable curve fitting to historical experimental data obtained during the culturing process of previous batches of the same cell culture. Based on the determination of the growth stage, the controller 26 can selectively signal the sonic array 24 to emit a predetermined sonication output ("programmed profile") that is suitable forthat growth stage. In some embodiments, each programmed profile has preselected sonication parameters ("profile parameters") for the predetermined sonication output. In some embodiments, the desired programmed profiles for various growth stages may be preprogrammed into the controller 26 by an operator of the bioreaction system 20. In additional or alternative embodiments, the programmed profiles may be based on user input from the operator to the controller 26, for example, via the user interface of the controller, at any time during the operation of the system 20.

[0048] In some embodiments, based on the signals generated by the transducers of the sonic array 24, the controller 26 can make continuous or periodic measurements that correlate to the real-time metabolite concentration of the cell culture CC in the tank 22. When the controller 26 determines that the metabolite concentration of the cell culture CC is at a peak, the controller 26 can selectively signal the operator of the bioreaction system 20 to begin extracting the cell culture CC from the tank 22 for downstream processing. A peak of the metabolite concentration does not necessarily mean that the metabolite concentration of the cell culture is at its absolute maximum level. In some embodiments, the peak may be where the yield isthe highest per culture time. In some embodiments, the controller 26 may determine whether the metabolite concentration of the cell culture CC is at a peak based, at least in part, on historical data from previous batches of the same cell culture.

[0049] In some embodiments, before extracting the cell culture CC from the tank 22 for downstream processing, the controller 26 may signal the sonic array 24 to sonicate at least a portion of the cell culture CC with sufficient acoustic energy to perform cavitation-based pasteurization of the cell culture CC. Pasteurization may be done in-situ or flow-through to reduce the microbial content of the cell culture CC to a desired level. In some embodiments, in- situ pasteurization is performed by one or more of the transducers 24a, 24b, 24c, 24d on the cell culture CC while the cell culture is in the tank 22.

[0050] In some embodiments, a transducer 24e of the sonic array 24 is configured to sonicate the cell culture as the cell culture flows past the transducer 24e. The transducer 24e is thus configured to perform "flow-through" sonication. In the illustrated embodiment, the transducer 24e is positioned at the outlet 28 of the tank 22 so that the transducer may impart sonication to the cell culture while the cell culture is exiting the tank 22. The transducer 24e may or may not be in direct contact with the cell culture to impart sonication thereto. In some embodiments, the transducer 24e penetrates the wall of the outlet 28 to directly contact the cell culture as the cell culture exits the tank 22. In other embodiments, the transducer 24e is in through-wall contact with the cell culture at the outlet 28.

[0051] In some embodiments, flow-through pasteurization is performed on the cell culture CC by operating the transducer 24e to sonicate the outgoing cell culture CC as the cell culture exits the tank 22, to thereby pasteurize the cell culture. In some embodiments, the flow rate of the cell culture CC at the outlet 28 is selectively controlled to allow sufficient sonic energy to be imparted to each volume of the cell culture exiting the tank 22, to achieve the desired level of pasteurization. [0052] In some embodiments, before the cell culture CC is extracted from the tank 22 for downstream processing, in-situ metabolite extraction is performed by the sonic array 24 on the cell culture CC while the cell culture is in the tank 22 or flow-through metabolite extraction is performed by transducer 24e of the sonic array 24 on the cell culture CC as the cell culture exits the tank 22 at outlet 28.

[0053] The bioreaction system 20 may be used to improve metabolite production in a suspended cell culture. In some embodiments, a method for improving metabolite production in a cell culture comprises determining, by the controller 26, a relative density of the cell culture in the tank 22 based on signals from sonic array 24 and, based on the relative density, adjusting the parameters of the sonication output emitted by the sonic array 24 as necessary, as the optimal sonication output for promoting metabolite production in the cell culture may vary for different relative densities of the cell culture.

[0054] In some embodiments, the determination of the relative density is based on real-time time-of-flight measurements that are taken periodically or continuously by the sonic array 24. In some embodiments, the relative density is a single spatially average value. In other embodiments, the relative density is a multi-dimensional image. In some embodiments, adjusting the parameters of the sonication output comprises modifying one or more of: a frequency, a power, a duty cycle, and a pulse width of the sonication output.

[0055] In some embodiments, the method comprises correlating the relative density to a growth stage of the cell culture CC. In some embodiments, the method comprises determining whether the cell culture CC is at a peak metabolite concentration; and upon determining that the cell culture CC is at a peak metabolite concentration, generating a signal to the operator of the bioreaction system 20. In some embodiments, the method comprises extracting the cell culture CC from the tank 22 for downstream processing. In some embodiments, the method comprises pasteurizing the cell culture CC, for example at the outlet of the tank 22, as the cell culture CC exits the tank 22.

[0056] In some embodiments, a method for improving metabolite production in a cell culture comprises determining, by the controller 26, a transmission attenuation of the cell culture in the tank 22 based on signals from sonic array 24 and, based on the transmission attenuation, adjusting the sonication output emitted by the sonic array 24 as necessary, as the optimal sonication output for promoting metabolite production in the cell culture may vary for different transmission attenuation of the cell culture.

[0057] In some embodiments, the transmission attenuation is a single spatially average value. In other embodiments, the transmission attenuation is a multi-dimensional image. In some embodiments, adjusting the sonication output comprises modifying one or more of: a frequency, a power, a duty cycle, and a pulse width of the sonication output.

[0058] In some embodiments, the method comprises correlating the transmission attenuation to a growth stage of the cell culture CC. In some embodiments, the method comprises determining whether the cell culture CC is at a peak metabolite concentration; and upon determining that the cell culture CC is at the peak metabolite concentration, generating a signal to the operator of the bioreaction system 20. In some embodiments, the method comprises extracting the cell culture CC from the tank 22 for downstream processing. In some embodiments, the method comprises pasteurizing the cell culture CC, for example at the outlet of the tank 22, as the cell culture CC exits the tank 22.

[0059] FIG. 3 shows a sample process 100 that can be carried out by the bioreaction system 20 according to some embodiments. The process 100 begins at step 102 where the sonic array 24 of the system 20 takes ultrasonic measurements of the cell culture CC in tank 22 and transmits corresponding signals to the controller 26. Based on the signals from the sonic array 24, the controller 26 performs in real-time one or more of the following actions: determine the relative density of the cell culture CC (step 104); determine the relative attenuation of the cell culture CC (step 106); determine the particle size distribution of the cell culture CC (step 108); and generate a multi-dimensional image of the relative attenuation or relative density of the cell culture CC (step 110).

[0060] At step 112, the controller 26 correlates the results from controller's actions at steps 104, 106, 108, and/or 110 to a growth stage of the cell culture CC. At step 114, after the controller determines the current growth stage, the controller 26 checks whether there is a programmed profile that corresponds to the current growth stage. If there is no programmed profile for the current growth stage, the process 100 returns to step 102 and continues to take ultrasonic measurements of the cell culture CC.

[0061] If there is a programmed profile for the current growth stage, the controller 26 signals the sonic array 24 to emit sonication having parameters in accordance with the programmed profile or to adjust the current sonication parameters to match those of the programmed profile (step 116). The sonic array 24 then emits sonication to the cell culture CC according to the signals from the controller 26 (step 118) and the process returns to step 102.

[0062] In some embodiments, simultaneously with or after step 114, the controller 26 may indicate to the operator, for example, via the display of the controller one or more of the following indicators with respect to the cell culture CC in real-time: yield, yield per time, harvest suitability, and metabolite concentration (step 120). Based on the indicators, the operator may selectively signal the system 20, for example by user input via the user interface of controller 26, to begin in-situ or flow-through pasteurization or metabolite extraction on the cell culture (step 122). Based on the operator's signal, the controller 26 sends signals to the sonic array 24 to begin pasteurization or metabolite extraction accordingly. In alternative embodiments, the controller 26 may automatically signal the sonic array 24 to begin in-situ or flow-through pasteurization or metabolite extraction on the cell culture based on the indicators (step 122). Based on the signals from the controller 26, the sonic array 24 sonicates the cell culture to perform pasteurization or metabolite extraction (step 124).

of Terms

[0063] Unless the context clearly requires otherwise, throughout the description and the "comprise", "comprising", and the like are to be construed in an inclusive sense, as opposed to an exclusive or exhaustive sense; that is to say, in the sense of "including, but not limited to"; "connected", "coupled", or any variant thereof, means any connection or coupling, either direct or indirect, between two or more elements; the coupling or connection between the elements can be physical, logical, or a combination thereof; "herein", "above", "below", and words of similar import, when used to describe this specification, shall refer to this specification as a whole, and not to any particular portions of this specification; "or", in reference to a list of two or more items, covers all of the following interpretations of the word: any of the items in the list, all of the items in the list, and any combination of the items in the list; the singular forms "a", "an", and "the" also include the meaning of any appropriate plural forms.

[0064] Where a component is referred to above, unless otherwise indicated, reference to that component should be interpreted as including as equivalents of that component any component which performs the function of the described component (/.e., that is functionally equivalent), including components which are not structurally equivalent to the disclosed structure which performs the function in the illustrated exemplary embodiments.

[0065] The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to those embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the full scope consistent with the claims. All structural and functional equivalents to the elements of the various embodiments described throughout the disclosure that are known or later come to be known to those of ordinary skill in the art are intended to be encompassed by the elements of the claims. Moreover, nothing disclosed herein is intended to be dedicated to the public regardless of whether such disclosure is explicitly recited in the claims. It is therefore intended that the following appended claims and claims hereafter introduced are interpreted to include all such modifications, permutations, additions, omissions, and sub-combinations as may reasonably be inferred. The scope of the claims should not be limited by the preferred embodiments set forth in the examples but should be given the broadest interpretation consistent with the description as a whole.

Claims

WHAT IS CLAIMED IS:

1. A bioreaction system for use on a cell culture, the bioreaction system comprising: a sonic array comprising two or more transducers, the sonic array being configured to selectively emit sonication, each of the two or more transducers being configured to be in direct or indirect contact with the cell culture, the sonication having sonication parameters; and a controller operably coupled to and in communication with the sonic array for selectively adjusting the sonication parameters and for receiving signals from the two or more transducers.

2. The bioreaction system of claim 1 wherein the controller is configured to determine or generate, based on the signals, one or more of: a density of the cell culture; an attenuation of the cell culture; a particle size distribution of the cell culture; and a multidimensional image of the cell culture.

3. The bioreaction system of claim 1 or 2 wherein the two or more transducers are spaced apart from one another.

4. The bioreaction system of any one of claims 1 to 3 wherein a plurality of the two or more transducers are configured to form a beam steering array.

5. The bioreaction system of any one of claims 1 to 4 wherein each of the two or more transducers is one or more of: a piezoelectric transducer, an electromagnetic transducer, and a magnetostrictive transducer.

6. The bioreaction system of any one of claims 1 to 5 wherein the controller comprises one or more of: a processer, a memory, a display, and a user interface.

7. The bioreaction system of any one of claims 1 to 6 wherein at least one of the two or more transducers is configured to perform flow-through sonication. The bioreaction system of any one of claims 1 to 7 comprising a tank for receiving the cell culture therein. The bioreaction system of claim 8 wherein the tank has a volume in a range from about 500L to about 5000L. The bioreaction system of claim 8 or 9 wherein a wall of the tank is made of a disposable membrane. A method comprising: taking ultrasonic measurements of a cell culture by two or more transducers of a sonic array, the two or more transducers being in direct or indirect contact with the cell culture; based on the ultrasonic measurements, performing one or more of: determining a particle size distribution of the cell culture; determining a relative attenuation of the cell culture; determining a relative density of the cell culture; and generating a multi-dimensional image of the cell culture; and correlating one or more of the particle size distribution, the relative attenuation, the relative density, and the multi-dimensional image to a growth stage of the cell culture. The method of claim 11 comprising, based on the growth stage, determining whether the cell culture is at a peak of metabolite concentration. The method of claim 12 comprising, upon determining that the cell culture is at the peak of metabolite concentration, generating a signal to an operator. The method of claim 11 comprising, based on one or more of the particle size distribution, the relative attenuation, the relative density, and the multi-dimensional image, providing an indicator for one or more of: a yield of the cell culture; a yield per culture time of the cell culture; a harvest suitability of the cell culture, and a metabolite concentration of the cell culture. The method of claim 11 comprising emitting sonication, by at least one of the two or more transducers, to the cell culture, the sonication having sonication parameters. The method of claim 15 comprising, based on the growth stage, adjusting at least one of the sonication parameters. The method of claim 15 comprising determining whether the growth stage has a corresponding programmed profile, the programmed profile having profile parameters; and upon determining that the growth stage has the corresponding programmed profile, causing the sonication parameters to match the profile parameters. The method of claim 15 comprising pasteurizing the cell culture by the at least one of the two or more transducers. The method of claim 18 wherein the pasteurizing is performed in-situ or flow-through. The method of claim 15 comprising performing metabolite extraction on the cell culture by the at least one of the two or more transducers. The method of claim 20 wherein the metabolite extraction is performed in-situ or flow- through. System having any new and inventive feature, combination of features, or subcombination of features as described herein. Method having any new and inventive steps, acts, combination of steps and/or acts or sub-combination of steps and/or acts as described herein.