Device and method for treating RNA with beads

Abstract

A fluidic device includes a first layer defining a first chamber portion configured to receive pressurized gas. The device also includes a second layer defining a second chamber portion positioned below the first chamber portion and configured to receive a liquid and a plurality of particles, a fluid outlet channel in fluid communication with the second chamber portion, and a filter interposed between the second chamber portion and the fluid outlet channel. The filter is formed in a surface of the second layer. The device also includes a resilient layer disposed between the first layer and the second layer, the resilient layer being deformable into the second chamber portion to drive the liquid out of the second chamber portion through the filter and into the fluid outlet channel. The filter is configured to allow the liquid to flow therethrough and prevent the particles from flowing therethrough.

Description

[0001] priority This application claims the benefit of U.S. Patent Application No. 63 / 551,768, filed February 9, 2024, entitled “Apparatus and Method for Treating RNA with Beads,” the disclosure of which is incorporated herein by reference.

[0002] background The topics discussed in this section should not be considered prior art simply because they are mentioned in this section. Similarly, problems mentioned in this section or related to the topics provided as background should not be considered as having been previously recognized in the prior art. The topics in this section represent only different methods, which in themselves may correspond to implementations of the claimed technology.

[0003] Some currently available technologies for manufacturing and formulating polynucleotide therapeutics (e.g., mRNA therapeutics) may expose the products to contamination and degradation. Some available centralized production facilities may be too expensive, too slow, or too susceptible to contamination for therapeutic formulations that may include multiple polynucleotide species. Brief description of the attached diagram Details of one or more embodiments are set forth in the accompanying drawings and the following description. Other features, aspects, and advantages will become apparent from the description, drawings, and claims, in which: Figure 1 A schematic diagram depicts an example of a system including a microfluidic processing chip; Figure 2 Depicting Figure 1 An exploded perspective view of instances of system components; Figure 3 It describes what can be incorporated into Figure 1 A top-view plan view of an instance of a processing chip in the system; Figure 4A Depicting the state in its first operation Figure 3 A cross-sectional side view of the processing chip; Figure 4B Depicting the second operating state Figure 3 A cross-sectional side view of the processing chip; Figure 4C Depicting the state in the third operation Figure 3 A cross-sectional side view of the processing chip; Figure 4D Depicting the fourth operating state Figure 3 A cross-sectional side view of the processing chip; Figure 4E Depicting the fifth operating state Figure 3 A cross-sectional side view of the processing chip; Figure 4F Depicting the sixth operating state Figure 3 A cross-sectional side view of the processing chip; Figure 5 Depicting merging Figure 1 A top perspective view of another exemplary processing chip in the system; Figure 6 Depicting Figure 5 Bottom perspective view of the processing chip; Figure 7 Depicting Figure 5 A top-down plan view of the processing chip, where some layers are transparent to show internal features; Figure 8 Depicting Figure 5 An exploded perspective view of the processing chip; Figure 9 Depicting Figure 5 Top perspective view of the first layer of the processing chip; Figure 10 Depicting Figure 9 Bottom perspective view of the first layer; Figure 11 Depicting Figure 9 First floor plan; Figure 12 Depicting Figure 5 A top plan view of the upper film of the second layer of the processing chip; Figure 13 Depicting Figure 5 A top plan view of the lower film of the second layer of the processing chip; Figure 14 Depicting Figure 5 Top perspective view of the third layer of the processing chip; Figure 15 Depicting Figure 14 The bottom perspective view of the third layer; Figure 16 Depicting Figure 14 Top plan of the third floor; Figure 17 Describing as Figure 16 shown Figure 14 Enlarged top plan view of area 17 on the third layer; Figure 18 Depicting Figure 14 A partial top perspective view of the third layer, showing the functionalized particulate filter; Figure 19 Describing as Figure 7 shown Figure 5An enlarged top plan view of region 19 of the processing chip, where some layers are transparent to show internal features, showing the flow path of functionalized particles through a portion of the processing chip; Figure 20 Depicting merging Figure 1 A schematic diagram of another example of a processing chip in a system; and Figure 21 Depicting merging Figure 1 A partial perspective view of another exemplary processing chip in the system.

[0005] Detailed description In some aspects, this document discloses apparatuses and methods for processing therapeutic polynucleotides. In particular, these apparatuses and methods can be closed-path apparatuses and methods configured to minimize or eliminate human handling during operation. Closed-path apparatuses and methods can provide a nearly completely sterile environment, and components can provide a sterile pathway for processing from an initial input (e.g., a template) to an output (e.g., a compound therapeutic agent). Materials input into the apparatus (e.g., nucleotides and any chemical components) can be sterile and can be introduced into the system with virtually no human interaction required.

[0006] The apparatus and methods described herein can be used to produce therapeutic agents with a high degree of reproducibility and rapid cycle times. The apparatus described herein can be configured to provide the synthesis, purification, dialysis, compounding, and concentration of one or more therapeutic compositions in a single integrated device. Alternatively, one or more of these processes can be performed in two or more devices as described herein. In some cases, the therapeutic composition may include a therapeutic polynucleotide, such as, for example, ribonucleic acid or deoxyribonucleic acid. The polynucleotide may consist only of natural nucleotide units, or may include any kind of synthetic, semi-synthetic, or modified nucleotide units. All or some of the processing steps can be performed in a continuous fluid processing pathway, which can be configured as one or a series of consumable microfluidic pathway devices, also referred to in some instances herein as processing chips or biochips (although the chip does not necessarily need to be used for biological applications). In some examples, the processing chip can be removably mounted in an instrument as part of a larger microfluidic system, such as… Figure 1 As shown in the diagram. The disclosed apparatus and method can be used to synthesize (including compound) patient-specific therapeutic agents at points of care (e.g., hospitals, clinics, pharmacies, etc.).

[0007] I. Terminology Throughout this specification and the appended claims, unless the context otherwise requires, the word “comprise” and variations such as “comprises” and “comprising” mean that various components / components may be used together in methods and articles (e.g., compositions and apparatuses and methods including devices). For example, the term “comprise” will be understood to imply the inclusion of any stated element or step, but does not exclude any other element or step. Generally, any apparatus and method described herein should be understood as inclusive, but all or a subset of components and / or steps may optionally be exclusive and may be expressed as “consisting of a plurality of components, steps, sub-components or sub-steps” or optionally “substantially consisting of a plurality of components, steps, sub-components or sub-steps”.

[0008] As used herein, unless the context clearly indicates otherwise, the singular forms “a”, “an”, and “the” are intended to include the plural forms as well. As used herein, the term “and / or” includes any and all combinations of one or more of the associated listed items and may be abbreviated to “ / ”.

[0009] Spatial terms such as “under,” “below,” “lower,” “over,” and “upper” are used herein to describe the relationship of one element or feature relative to another element or feature, or multiple elements or features, as shown in the accompanying drawings. It will be understood that spatially related terms are intended to include different orientations of the device in use or operation, other than those depicted in the drawings. For example, if the device in the drawings is inverted, an element described as “below” or “under” other elements or features would be oriented “above” other elements or features. Thus, the term “under” can include both above and below orientations. The device may be oriented otherwise (rotated 90 degrees or otherwise), and the spatially related descriptive terms used herein are interpreted accordingly. Similarly, terms such as “upwardly,” “downwardly,” “vertical,” and “horizontal” are used herein for illustrative purposes only, unless otherwise specifically stated.

[0010] When a feature or element is referred to herein as being “on” another feature or element, it may be directly on the other feature or element, or there may be an intermediate feature and / or element. Conversely, when a feature or element is referred to as being “directly on” another feature or element, there is no intermediate feature or element. When a feature or element is referred to as being “connected,” “attached,” or “coupled” to another feature or element, the feature or element may be directly connected, attached, or coupled to the other feature or element, or there may be an intermediate feature or element. Conversely, when a feature or element is referred to as being “directly connected,” “directly attached,” or “directly coupled” to another feature or element, there is no intermediate feature or element. While one embodiment has been described or illustrated, the features and elements thus described or illustrated can be applied to other embodiments. Those skilled in the art will also recognize that references to structures or features configured to be “adjacent” to another feature may have portions overlapping with or below the adjacent feature.

[0011] As used herein in the specification and claims, including in the examples, and unless expressly stated otherwise, all figures are to be interpreted as if preceded by the words “about” or “approximately,” even if the term is not explicitly stated. The phrase “about” or “approximately” may be used when describing a quantity and / or location to indicate that the described value and / or location is within a reasonably expected range of the value and / or location. For example, a numerical value may have values ​​of ±0.1% of the stated value (or range of values), ±1% of the stated value (or range of values), ±2% of the stated value (or range of values), ±5% of the stated value (or range of values), ±10% of the stated value (or range of values), etc. Any numerical value given herein should also be understood to include about or approximately that value, unless the context otherwise requires. For example, if the value “10” is disclosed, “about 10” is also disclosed. Any numerical ranges listed herein are intended to include all subranges contained therein.

[0012] It should also be understood that when a value is disclosed, "less than or equal to" that value, "greater than or equal to" that value, and possible ranges between the values ​​are also disclosed, as would be appropriately understood by a person skilled in the art. For example, if the value "X" is disclosed, then "less than or equal to X" and "greater than or equal to X" (e.g., where X is a numerical value) are also disclosed. It should also be understood that throughout the application, data is provided in a variety of different formats, and that the data represents endpoints and starting points, as well as ranges for any combination of data points. For example, if a specific data point "10" and a specific data point "15" are disclosed, then it should be understood that greater than, greater than or equal to, less than, less than or equal to, and equal to 10 and 15, as well as ranges between 10 and 15, are considered to be disclosed. It should also be understood that each unit between two specific units is also disclosed. For example, if 10 and 15 are disclosed, then 11, 12, 13, and 14 are also disclosed.

[0013] While the terms "first" and "second" may be used herein to describe various features / elements (including steps), these features / elements should not be limited by these terms unless the context otherwise requires. These terms are used to distinguish one feature / element from another and do not indicate a particular order unless specifically stated. Therefore, without departing from the teachings of the invention, the first feature / element discussed below may be referred to as the second feature / element, and similarly, the second feature / element discussed below may be referred to as the first feature / element.

[0014] As used herein, the terms “system,” “apparatus,” and “equipment” are to be understood as interchangeable. A system, apparatus, and equipment may each comprise more than one component having various structural and / or functional relationships with each other.

[0015] As used herein, "polynucleotide" refers to a nucleic acid molecule containing multiple nucleotides, and generally refers to both "oligonucleotide" (a polynucleotide molecule of 18-25 nucleotides in length) and polynucleotides of 26 or more nucleotides. Aspects of this disclosure include compositions comprising: oligonucleotides of 18-25 nucleotides in length (e.g., 18-mer, 19-mer, 20-mer, 21-mer, 22-mer, 23-mer, 24-mer, or 25-mer), or intermediate-length polynucleotides of 26 or more nucleotides in length (e.g., 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 3 ...9, 39, 39, 39, 39, 39, 39, 39, 39, 39, 39, 39, 39, 39, 39, 3 9, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, approximately 65, approximately 70, approximately 75, approximately 80, approximately 85, approximately 90, approximately 95, approximately 100, approximately 110, approximately 120, approximately 130, approximately 140, approximately 150, approximately 160, approximately 170, approximately 180, approximately 190, approximately 200, approximately 210, approximately 220, approximately 230, approximately 24 Polynucleotides of about 0, about 250, about 260, about 270, about 280, about 290, or about 300 nucleotides, or long polynucleotides longer than about 300 nucleotides (e.g., between about 300 and about 400 nucleotides, between about 400 and about 500 nucleotides, between about 500 and about 600 nucleotides, between about 600 and about 700 nucleotides, between about 700 and about 800 nucleotides, between about 800 and about 900 nucleotides, between about 900 and about 300 nucleotides). Polynucleotides can be categorized into lengths of approximately 1000 nucleotides, approximately 300 to approximately 500 nucleotides, approximately 300 to approximately 600 nucleotides, approximately 300 to approximately 700 nucleotides, approximately 300 to approximately 800 nucleotides, approximately 300 to approximately 900 nucleotides, or approximately 1000 nucleotides, or even longer than approximately 1000 nucleotides, such as polynucleotides of approximately 1500, approximately 2000, approximately 2500, approximately 5000, or approximately 7000 nucleotides. In the case of a double-stranded polynucleotide, its length can be similarly described using base pairs.

[0016] As used herein, “amplification” can refer to polynucleotide amplification. Amplification can include any suitable method for amplifying polynucleotides, and includes, but is not limited to, multiple displacement amplification (MDA), polymerase chain reaction (PCR) amplification, loop-mediated isothermal amplification (LAMP), nucleic acid sequence-based amplification, strand displacement amplification, rolling circle amplification, and ligase chain reaction.

[0017] As used herein, a “cassette” (e.g., a synthetic in vitro transcription promoter cassette) refers to a polynucleotide sequence that may include or be operatively linked to one or more expression elements, such as enhancers, promoters, leader sequences, introns, 5' untranslated regions (UTRs), 3' UTRs, or transcription termination sequences. In some aspects, a cassette contains at least a first polynucleotide sequence capable of initiating transcription of an operatively linked second polynucleotide sequence (which may contain a template) and optionally a transcription termination sequence operatively linked to the second polynucleotide sequence. As described below, the template may contain a sequence of interest, such as an open reading frame of interest (“ORF”). Cassettes may be provided as a single element or as two or more unlinked elements.

[0018] As used herein, a “template” refers to a nucleic acid sequence containing a sequence of interest for preparing a therapeutic polynucleotide according to the disclosed methods. A template can be, but is not limited to, double-stranded DNA (dsDNA), engineered plasmid constructs, cDNA sequences, or linear nucleic acid sequences (e.g., linear templates generated by PCR or by annealing chemistry of oligonucleotides). In some respects, the template can be integrated into a “box” as described above.

[0019] As used herein, the term “sequence of interest” refers to a polynucleotide sequence whose use is considered desirable for a suitable purpose, particularly for the preparation of mRNA for therapeutic purposes; and includes, but is not limited to, coding sequences of structural genes and non-coding regulatory sequences that do not encode mRNA or protein products.

[0020] As used herein, “in vitro transcription” or “IVT” refers to the process by which transcription occurs in vitro in a non-cellular system to produce synthetic RNA molecules (e.g., synthetic mRNA) for various applications, including therapeutic delivery to a subject, such as as therapeutic polynucleotides, which may be part of or used to form therapeutic polynucleotide compositions as described below. The resulting therapeutic polynucleotides (e.g., synthetic RNA molecules (transcription products)) may be combined with a delivery medium to form therapeutic polynucleotide compositions. Synthetic transcription products include mRNA, antisense RNA molecules, shRNA, circular RNA molecules, ribozymes, etc. IVT reactions may use a purified linear DNA template containing a promoter sequence and an open reading frame (ORF) sequence of interest, ribonucleotide triphosphates or modified ribonucleotide triphosphates, a buffer system including DTT and magnesium ions, and phage RNA polymerase.

[0021] As used herein, "therapeutic polynucleotide" means a polynucleotide (e.g., mRNA) that may be part of a therapeutic polynucleotide composition intended for delivery to a subject to treat the subject's symptoms, disease, or condition; prevent the subject's symptoms, disease, or condition; or improve or otherwise alter the subject's health.

[0022] As used herein, a “therapeutic polynucleotide composition” (or simply “therapeutic composition”) can refer to a composition comprising one or more therapeutic polynucleotides (e.g., mRNA) encapsulated by a delivery medium, which can be administered to a subject in need using any suitable route of administration, such as intratumoral, intramuscular, or other injection. An example of a therapeutic polynucleotide composition is mRNA (therapeutic) nanoparticles comprising at least one mRNA encapsulated by a delivery medium molecule. mRNA vaccines are an example of therapeutic polynucleotide compositions.

[0023] As used herein, “delivery vehicle” means any substance that at least partially facilitates the delivery of polynucleotides (e.g., therapeutic polynucleotides) in vivo, in vitro, or ex vivo to target cells or tissues (e.g., tumors, etc.). Referring to something as a delivery vehicle does not preclude the possibility that the delivery vehicle may also have therapeutic effects. Some versions of delivery vehicles may provide additional therapeutic effects. In some versions, the delivery vehicle may be a peptide-like molecule, such as an aminoesterified peptide molecule, which can be used to at least partially encapsulate mRNA. The term “DV” will also be used herein as an abbreviation for “delivery vehicle.”

[0024] As used in this article, “linking” refers to methods used to couple one component to another, such as linking, synthesis, primer extension, annealing, recombination, or hybridization.

[0025] As used herein, “purification” refers to the physical and / or chemical separation of a component (e.g., particles) from other unwanted components (e.g., contaminants, debris, etc.).

[0026] As used herein, the term “substantially free” with respect to a given substance includes 100% free of the given substance, or containing less than about 1.0%, or less than about 0.5%, or less than about 0.1% of the given substance.

[0027] II. Overview of Systems Including Microfluidic Processing Chips Figure 1Examples of various components that can be incorporated into the system (100) are depicted. The system (100) of this example includes a housing (103) that surrounds a seating mount (115) capable of removably holding one or more microfluidic processing chips (111). In other words, the system (100) includes a chip receiving component configured to removably house the processing chip (111), wherein the processing chip (111) itself defines one or more microfluidic channels or fluid pathways. Components of the system (100) that fluidly interact with the processing chip (111) (e.g., within the housing (103)) may include fluid channels or pathways that are not necessarily considered microfluidics (e.g., such fluid channels or pathways are larger than the microfluidic channels or fluid pathways in the processing chip (111)). In some versions, the processing chip (111) is provided and used as a disposable device; while the rest of the system (100) is reusable. The housing (103) may be in the form of a chamber, shell, etc., having openings that can be closed (e.g., closed via a cover or door, etc.) to seal the interior. The housing (103) may enclose the thermal regulator and / or may be configured to be enclosed in a thermally regulated environment (e.g., a refrigeration unit, etc.). The housing (103) may form a sterile barrier. In some variations, the housing (103) may form a humidified or humidity-controlled environment. Alternatively, the system (100) may be positioned in a cabinet (not shown). Such a cabinet may provide a temperature-regulated (e.g., refrigerated) environment. Such a cabinet may also provide air filtration and airflow management and may help maintain reagents at the desired temperature throughout the manufacturing process. Additionally, such a cabinet may be equipped with UV lamps for sterilizing the processing chip (111) and other components of the system (100). Other suitable features may be incorporated into the cabinet housing the system (100).

[0028] In some cases, an assembly formed by a housing (103) and components of a system (100) within the housing (103) (without a processing chip (111)) can be considered an "instrument". While the controller (121) and user interface (123) are in Figure 1The controller (121) and user interface (123) are shown outside the housing (103), but the controller (121) and user interface (123) can actually be located inside or on the housing (103) and thus can also form part of the instrument. As described in more detail below, the instrument can removably receive the processing chip (111) via a mount (115). When the processing chip (111) is placed in the mount (115), the instrument and the processing chip (111) cooperate to form the system (100) together. When the processing chip (111) is removed from the mount (115), the remaining portion of the system (100) can be considered as the “instrument”. The instrument, the system (100) and the processing chip (111) can each be considered as a “device”. Thus, the term “device” can be understood to include the instrument itself, the processing chip (111) itself, combinations of the instrument and the processing chip (111), some other combination of components of the system (100), or some other arrangement of the system (100) or its components.

[0029] The mounting bracket (115) can be configured to hold the processing chip (111) in a fixed and predefined orientation using one or more pins or other components configured to hold the processing chip (111) in a fixed and predefined orientation. Therefore, the mounting bracket (115) facilitates holding the processing chip (111) in a proper position and orientation relative to other components of the system (100). In this example, the mounting bracket (115) is configured to hold the processing chip (111) in a horizontal orientation such that the processing chip (111) is parallel to the ground.

[0030] In some variations, the thermal control (113) may be located near the mount (115) to regulate the temperature of any processing chip (111) mounted on the mount (115). The thermal control (113) may include thermoelectric components (e.g., a Peltier device) and / or one or more heat sinks for controlling all or part of the temperature of any processing chip (111) mounted on the mount (115). In some variations, more than one thermal control (113) may be included, for example, to individually regulate the temperature of different areas in one or more regions of the processing chip (111). The thermal control (113) may include one or more thermal sensors (e.g., thermocouples) that can be used for feedback control of the processing chip (111) and / or the thermal control (113).

[0031] like Figure 1As shown, the fluid interface assembly (109) couples the processing chip (111) to a pressure source (117), thereby providing one or more pathways for delivering positive or negative pressure fluid (e.g., gas) from the pressure source (117) to one or more internal regions of the processing chip (111), as will be described in more detail below. Although only one pressure source (117) is shown, the system (100) may include two or more pressure sources (117). In some cases, pressure may be generated by one or more sources other than the pressure source (117). For example, one or more vials or other fluid sources within the reagent storage frame (107) may be pressurized. Alternatively, reactions and / or other processes performed on the processing chip (111) may generate additional fluid pressure. In this example, the fluid interface component (109) also couples the processing chip (111) to the reagent storage frame (107), thereby providing one or more paths for liquid reagents, etc., to be transferred from the reagent storage frame (107) to one or more internal areas of the processing chip (111), as will be described in more detail below.

[0032] In some versions, pressurized fluid (e.g., gas) from at least one pressure source (117) reaches the fluid interface assembly (109) via a reagent storage frame (107), such that the reagent storage frame (107) includes one or more components inserted into the fluid path between the pressure source (117) and the fluid interface assembly (109). In some versions, one or more pressure sources (117) are directly coupled to the fluid interface assembly, such that positively pressurized fluid (e.g., positively pressurized gas) or negatively pressurized fluid (e.g., suction or other negatively pressurized gas) bypasses the reagent storage frame (107) to reach the fluid interface assembly (109). Regardless of whether the fluid interface assembly (109) is inserted into the fluid path between the pressure source (117) and the fluid interface assembly (109), the fluid interface assembly (109) can be removably coupled to the remainder of the system (100), such that at least a portion of the fluid interface assembly (109) can be removed for sterilization between uses. As described in more detail below, the pressure source (117) can selectively pressurize one or more chamber regions on the processing chip (111). Alternatively, the pressure source can also selectively pressurize one or more vials or other fluid storage containers held by the reagent storage frame (107).

[0033] The reagent storage frame (107) is configured to include more than one fluid sample holder, each of which can hold a fluid vial configured to store reagents (e.g., nucleotides, solvents, water, etc.) for delivery to the processing chip (111). In some versions, one or more fluid vials or other storage containers in the reagent storage frame (107) may be configured to receive products from the interior of the processing chip (111). Alternatively, a second processing chip (111) may receive products from the interior of a first processing chip (111), such that one or more fluids are transferred from one processing chip (111) to another. In some such cases, the first processing chip (111) may perform a first dedicated function (e.g., synthesis, etc.), while the second processing chip (111) performs a second dedicated function (e.g., encapsulation, etc.). The reagent storage frame (107) of this example includes more than one pressure line and / or manifold configured to divide one or more pressure sources (117) into more than one pressure line that can be applied to the processing chip (111). Such pressure lines can be controlled independently or collectively (in the form of sub-combinations).

[0034] The fluid interface assembly (109) may include more than one fluid line and / or pressure line, each of which includes a biased (e.g., spring-loaded) retainer or tip that individually and independently drives each fluid line and / or pressure line to the processing chip (111) when the processing chip (111) is held in the holder (115). Any associated tubing (e.g., fluid lines and / or pressure lines) may be part of and / or connected to the fluid interface assembly (109). In some versions, each fluid line includes a flexible tube connected between the reagent storage frame (107) and the processing chip (111) via a connector that couples the vial to the tube in a locking engagement (e.g., a collar). In some versions, the ends of the fluid / pressure lines can be configured to seal against the processing chip (111) (e.g., at corresponding sealing ports formed in the processing chip (111)), as described below. In this example, when the processing chip (111) is in place in the holder (115), the connection between the pressure source (117) and the processing chip (111), and the connection between the vial in the reagent storage frame (107) and the processing chip (111), form isolated, sealed, and closed pathways. These sealed, closed pathways provide protection against contamination when processing therapeutic polynucleotides.

[0035] The vials in the reagent storage frame (107) can be pressurized (e.g., pressures >1 atm, such as 2 atm, 3 atm, 5 atm, or higher). In some versions, the vials can be pressurized via a pressure source (117). Negative or positive pressure can thus be applied. For example, fluid vials can be pressurized to between about 1 psig and about 20 psig (e.g., 5 psig, 10 psig, etc.). Alternatively, at the end of the treatment, a vacuum (e.g., about -7 psig or about 7 psia) can be applied to aspirate the fluid back into the vial (e.g., a vial used as a depot). The fluid vials can be actuated at a lower pressure than the pneumatic valve, as described below, which can prevent or reduce leakage. In some variations, the pressure difference between the fluid and the pneumatic valve can be between about 1 psi and about 25 psi (e.g., about 3 psi, about 5 psi, 7 psi, 10 psi, 12 psi, 15 psi, 20 psi, etc.).

[0036] The system (100) of this example also includes a magnetic field applicator (119) configured to generate a magnetic field at a region of the processing chip (111). The magnetic field applicator (119) may include a movable head operable to move the magnetic field, thereby selectively isolating products adhering to the magnetic trap beads within other storage containers in vials or reagent storage frames (107).

[0037] The system (100) of this example also includes one or more sensors (105). In some versions, such sensors (105) include one or more cameras and / or other types of optical sensors. Such sensors (105) can sense one or more of the following: a barcode, a liquid level in a vial held within a reagent storage frame (107), fluid movement within a processing chip (111) mounted within a mount (115), and / or other optically detectable conditions. In versions where the sensor (105) is used to sense a barcode, such a barcode may be included on a vial in the reagent storage frame (107), allowing the sensor (105) to identify the vial in the reagent storage frame (107). In some versions, a single sensor (105) is positioned and configured to simultaneously view such a barcode on a vial in the reagent storage frame (107), a liquid level in a vial in the reagent storage frame (107), fluid movement within a processing chip (111) mounted within a mount (115), and / or other optically detectable conditions. In some other versions, more than one sensor (105) is used to view these conditions. In some such versions, different sensors (105) can be positioned and configured to view the corresponding optically detectable conditions individually, so that the sensor (105) can be dedicated to a specific corresponding optically detectable condition.

[0038] In versions of the sensor (105) that include at least one optical sensor, visual / optical markers can be used to estimate yield. For example, fluorescence can be used to detect processing yields or residues by labeling with fluorophores. Alternatively, dynamic light scattering (DLS) can be used to measure particle size distribution within a portion of the processing chip (111), such as a mixing portion of the processing chip (111). In some variations, the sensor (105) can use one or two optical fibers to deliver light (e.g., laser) to the processing chip (111) to provide measurement results; and to detect optical signals emanating from the processing chip (111). In versions where the sensor (105) optically detects processing yields or residues, the sensor (105) can be configured to detect visible light, fluorescence, ultraviolet (UV) absorbance signals, infrared (IR) absorbance signals, and / or any other suitable type of optical feedback.

[0039] In versions where the sensor (105) includes at least one optical sensor configured to capture video images, such a sensor (105) can record at least some activity on the processing chip (111). For example, the entire process (run) for synthesizing and / or processing a substance (e.g., therapeutic RNA) can be recorded by one or more video sensors (105), including those capable of visualizing the processing chip (111) from above. The processing on the processing chip (111) can be visually tracked, and the video recording can be retained for subsequent quality control and / or processing. Thus, the video recording of the processing can be saved, stored, and / or transmitted for subsequent review and / or analysis. Additionally, as will be described in more detail below, the video can be used as a real-time feedback input that can influence processing using at least visually observable conditions captured in the video.

[0040] The system (100) of this example can be controlled by a controller (121). The controller (121) may include one or more processors, one or more memories, and various other suitable electrical components. In some versions, one or more components of the controller (121) (e.g., one or more processors, etc.) are embedded within the system (100) (e.g., contained within a housing (103)). Alternatively, one or more components of the controller (121) (e.g., one or more processors, etc.) may be detachably attached to or detachably connected to other components of the system (100). Thus, at least a portion of the controller (121) may be removable. Furthermore, in some versions, at least a portion of the controller (121) may be remote from the housing (103).

[0041] In addition to other tasks, the control of the controller (121) may also include activating a pressure source (117) to apply pressure via the processing chip (111) to drive fluid motion. The controller (121) may be wholly or partially outside the housing (103); or wholly or partially inside the housing (103). The controller (121) may be configured to receive user input via a user interface (123) of the system (100); and to provide output to the user via the user interface (123). In some versions, the controller (121) is fully automated to the point that no user input is required. In some such versions, the user interface (123) may only provide output to the user. The user interface (123) may include a monitor, a touchscreen, a keyboard, and / or any other suitable feature. The controller (121) can coordinate processes including: moving one or more fluids onto or on the processing chip (111), mixing one or more fluids on the processing chip (111), adding one or more components to the processing chip (111), metering the fluids in the processing chip (111), adjusting the temperature of the processing chip (111), and applying a magnetic field (e.g., when using a magnetic bead). The controller (121) can receive real-time feedback from a sensor (105) and execute a control algorithm based on such feedback. Such feedback from the sensor (105) may include, but is not limited to, identification of reagents in vials in a reagent storage frame (107), detected liquid levels in vials in a reagent storage frame (107), detected movement of fluids in the processing chip (111), fluorescence of fluorophores in the fluids in the processing chip (111), etc. The controller (121) may include software, firmware, and / or hardware. The controller (121) can also communicate with a remote server, for example, to track the operation of the device, reorder materials (e.g., components such as nucleotides, processing chips (111) etc.) and / or download protocols, etc.

[0042] Figure 2 Examples of specific forms that the various components of the system (100) can take are shown. In particular, Figure 2The reagent storage frame (150), fluid interface assembly (152), stand (154), thermal control (156), and processing chip (200) are shown. The reagent storage frame (150), fluid interface assembly (152), stand (154), thermal control (156), and processing chip (200) of this example can be configured and operated as described above for the reagent storage frame (107), fluid interface assembly (109), stand (115), thermal control (113), and processing chip (111), respectively. These components are fixed relative to a base (180). A set of rods (182) supports the reagent storage frame (150) above the fluid interface assembly (152).

[0043] like Figure 2 As shown, a set of optical sensors (160) are positioned at four corresponding locations along the base (180). The optical sensors (160) can be configured and operated as described above with the sensor (105). The optical sensors (160) can include off-the-shelf cameras or any other suitable type of optical sensor. The optical sensors (160) are positioned such that the fluid vials held within the reagent storage frame (150) are within the field of view of one or more optical sensors (160). Additionally, the processing chip (200) is within the field of view of one or more optical sensors (160). Each optical sensor (160) is movably fixed to the base (180) via a corresponding track (184) (e.g., in a gantry arrangement), such that each optical sensor (160) is configured to translate laterally along each corresponding track (184). A linear actuator (186) is attached to each optical sensor (160) and thereby operable to drive each optical sensor (160) to lateral translation along a corresponding track (184). Each actuator (186) may be in the form of a drive belt, drive chain, drive cable, or any other suitable type of structure. A controller (121) may drive the operation of the actuators (186). The optical sensors (160) may move along the track (184) during operation of the system (100) to view appropriate areas of vials and / or processing chips (200) in the reagent storage frame (150). In some cases, the optical sensors (160) move uniformly along their corresponding tracks (184). In other cases, the optical sensors (160) move independently along their corresponding tracks (184).

[0044] Although the optical sensor (160) is in Figure 2The optical sensor (160) is shown mounted to a base (180), but in addition to mounting to a base (180), or as an alternative to mounting to a base (180), the optical sensor (160) may be positioned at other locations within the system (100). For example, some versions of the reagent storage frame (107) may include one or more optical sensors (160) positioned and configured to provide an overhead field of view. In some such versions, such optical sensors (160) may be mounted to a track, a movable cantilever, or other structure that allows such optical sensors (160) to be repositioned during operation of the system (100). The optical sensor (160) may be positioned at any other suitable location. Although not shown, the system (100) may also include one or more light sources (e.g., electroluminescent panels, etc.) to provide illumination that aids in the optical sensing of the optical sensor (160).

[0045] In some versions, one or more mirrors are used to facilitate the visualization of components of the system (100) through an optical sensor (160). Such mirrors allow the optical sensor (160) to view components of the system (100) that might otherwise not be within the sensor's field of view. These mirrors can be placed directly adjacent to the optical sensor (160). Alternatively, such mirrors can be placed adjacent to one or more components of the system (100) that are to be viewed through the optical sensor (160).

[0046] When using the system (100), the operator can select a protocol to run (e.g., from a library of preset protocols), or the user can input a new protocol (or modify an existing one) via a user interface (123). Depending on the protocol, the controller (121) can instruct the operator: which processing chip (111) to use, what the contents of the vials in the reagent storage frame (107) should be, and where to place the vials within the reagent storage frame (107). The operator can load the processing chip (111) into the holder (115); and load the desired reagent vials and export vials into the reagent storage frame (107). The system (100) can confirm the presence of the desired peripheral device, identify the processing chip (111), and scan the identifier (e.g., barcode) of each reagent and product vial in the reagent storage frame (107) to match the vials with the bill-of-reagent for the selected protocol. After confirming the starting materials and equipment, the controller (121) can execute the protocol. During execution, as described in more detail below, valves and pumps are actuated to deliver reagents, mix reagents, control temperature, conduct reactions, perform measurements, and pump the products to the target vial in the reagent storage frame (107).

[0047] III. Processing Chip Examples Figure 3 and Figures 4A-4F An example of the processing chip (200) is described in more detail. In conjunction with the rest of the system (100), the processing chip (200) can be used to provide the in vitro synthesis, purification, concentration, formulation, and / or analysis of therapeutic compositions, including but not limited to therapeutic polynucleotides and therapeutic polynucleotide compositions. Figure 3 As shown, the processing chip (200) of this example includes more than one fluid port (220). Each fluid port (220) has an associated fluid channel (222) formed in the processing chip (200) such that fluid delivered to the fluid port (220) will flow through the corresponding fluid channel (222). As described in more detail below, each fluid port (220) is configured to receive fluid from a corresponding fluid line (206) from the fluid interface assembly (109). In this example, each fluid channel (222) leads to a valve chamber (224) operable to selectively block or allow fluid from the corresponding fluid channel (222) to be further conveyed along the processing chip (200), as will be described in more detail below.

[0048] In addition, such as Figure 3As shown, the processing chip (200) of this example includes more than one additional chamber (230, 250, 270) which can be used to serve different purposes during the process of producing a therapeutic composition, as described herein. By way of example only, such additional chambers (230, 250, 270) can be used to provide the synthesis, purification, dialysis, compounding, and concentration of one or more therapeutic compositions; or to perform any other suitable function. Fluid can be transferred from one chamber (230) to another chamber (230) via a fluid connector (232). In some versions, the fluid connector (232) can operate like a valve between an open and closed state (e.g., similar to a valve chamber (224)). In some other versions, the fluid connector (232) remains open throughout the manufacture of the therapeutic composition. In this example, chamber (230) is used to provide the synthesis of polynucleotides, although chamber (230) can alternatively be used for any other suitable purpose.

[0049] exist Figure 3 In the example shown, another valve chamber (234) is inserted between one chamber (230) and one chamber (250), allowing fluid to be selectively transferred from chamber (230) to chamber (250). The chambers (250) are provided in pairs and coupled to each other, allowing the processing chip (200) to transfer fluid back and forth between the chambers (250). While a pair of chambers (250) is provided in this example, any other suitable number of chambers (250) can be used, including only one chamber (250) or more than two chambers (250). The chambers (250) can be used to provide purification of the fluid and / or for any of the various other purposes described herein; and can have any suitable configuration. In versions of the chamber (250) used for purification, the chamber (250) may include material configured to absorb selected portions of the fluid mixture in the chamber (250). In some such versions, the material may include a cellulose material that can selectively absorb double-stranded mRNA from the mixture. In some such versions, the cellulose material can be inserted into only one of a pair of chambers (250), such that when fluid is mixed from the first chamber (250) to the second chamber (250) of the pair, mRNA and / or some other components can be efficiently removed from the fluid mixture, which can then be transferred downstream to another pair of chambers (270) for further processing or output. Alternatively, the chambers (250) can be used for any other suitable purpose.

[0050] An additional valve chamber (252) is inserted between each chamber (250) and its corresponding chamber (270), allowing fluid to be selectively transferred from chamber (250) to chamber (270) via the valve chamber (252). The chambers (270) are also coupled to each other, allowing the processing chip (200) to transfer fluid back and forth between the chambers (270). The chambers (270) can be used to provide mixing of fluids and / or for any of the various other purposes described herein; and can have any suitable configuration.

[0051] like Figure 3 As shown, chamber (270) is also coupled to additional fluid ports (221) via corresponding fluid channels (223) and valve chambers (225). The fluid ports (221), fluid channels (223), and valve chambers (225) can be configured and operated as described above for fluid ports (220), fluid channels (222), and valve chambers (224). In some versions, the fluid port (221) is used to transfer additional fluid to chamber (270). Alternatively, the fluid port (221) can be used to transfer fluid from processing chip (200) to another device. For example, fluid from chamber (270) can be transferred directly via fluid port (221) to another processing chip (200), to one or more vials in reagent storage frame (107), or elsewhere.

[0052] The processing chip (200) also includes several storage chambers (260). In this example, each storage chamber (260) is configured to receive and store fluid being transferred to or from a corresponding chamber (250, 270). Each storage chamber (260) has a corresponding inlet valve chamber (262) and an outlet valve chamber (264). Each inlet valve chamber (262) is inserted between the storage chamber (260) and the corresponding chamber (250, 270) and is thereby operable to allow or prevent fluid flow between the storage chamber (260) and the corresponding chamber (250, 270). Each outlet valve chamber (264) is operable to meter the fluid flow rate between the storage chamber (260) and the corresponding fluid port (266). In some versions, each fluid port (266) is configured to transfer fluid from a corresponding vial in a reagent storage frame (107) to the corresponding storage chamber (260). Alternatively, each fluid port (266) may be configured to transfer fluid from the corresponding reservoir (260) to the corresponding vial in the reagent storage frame (107). In this example, the reservoir (260) is used to provide metering of fluid transferred to and / or from the processing chip (200). Alternatively, the reservoir (260) may be used for any other suitable purpose, including but not limited to pressurizing fluid transferred to and / or from the processing chip (400).

[0053] For example Figure 3 As shown, the processing chip (200) of this example includes more than one pressure port (240). Each pressure port (240) has an associated pressure channel (244) formed in the processing chip (200) such that pressurized gas delivered through the pressure port (240) will be further delivered through the corresponding pressure channel (244). As described in more detail below, each pressure port (240) is configured to receive pressurized gas from a corresponding pressure line (208) from the fluid interface assembly (109). In this example, each pressure channel (244) leads to a corresponding chamber (224, 225, 230, 234, 250, 252, 260, 262, 264, 270) thereby providing valve regulation or peristaltic pumping via these chambers (224, 225, 230, 234, 250, 252, 260, 262, 264, 270), as described in more detail below.

[0054] The processing chip (200) may also include electrical contacts, pins, pin sockets, capacitor coils, induction coils, or other features configured to provide electrical communication with other components of the system (100). Figure 3 In the example shown, the processing chip (200) includes an electrically active region (212) that includes electrical communication features. The electrically active region (212) may also include circuitry and other electrical components. In some versions, the electrically active region (212) may provide communication of power, data, etc. Although the electrically active region (212) is shown at a specific location on the processing chip, it may alternatively be located at any other suitable location. In some versions, the electrically active region (212) is omitted.

[0055] like Figures 4A-4F As shown, the processing chip (200) also includes a first plate (300), an elastic layer (302), a second plate (304), and a third plate (306). As described in more detail below, some versions of the elastic layer (302) are in the form of a flexible film. The first plate (300) has an upper surface (210) and a lower surface (310), the lower surface (310) being opposite to the elastic layer (302). The second plate (304) has an upper surface (312) and a lower surface (314), the upper surface (312) being opposite to the elastic layer (302); and the lower surface (314) being opposite to the third plate (306). Thus, the elastic layer (302) is inserted between the first plate (300) and the second plate (304). In this example, another elastic layer (316) is also inserted between the second and third plates (304, 306), although this elastic layer (316) is optional.

[0056] The plates (300, 304, 306) in this example are substantially translucent to visible and / or ultraviolet light. "Substantially translucent" means that at least 90% (in some cases, including 100%) of the light is transmitted through the material compared to a translucent material. In some variations, one or more of the plates (300, 304, 306) may comprise a material substantially transparent to visible and / or ultraviolet light. "Substantially transparent" means that at least 90% (in some cases, including 100%) of the light is transmitted through the material compared to a completely transparent material. As another example, one or more of the plates (300, 304, 306) may provide transmission of ultraviolet light at a wavelength of about 260 nm, with a transmittance ranging from about 0.2% to about 20%, including from about 0.4% to about 15%, or including from about 0.5% to about 10%.

[0057] The plates (300, 304, 306) in this example are also rigid. In some other versions, one or more of the plates (300, 304, 306) are semi-rigid. The plates (300, 304, 306) may include glass, plastic, silicone, and / or any other suitable material. In some versions, one or more of the plates (300, 304, 306) are formed as a laminate of two or more layers of material, such that each plate (300, 304, 306) does not necessarily need to be formed as a single homogeneous continuous material. The material constituting one of the plates (300, 304, 306) may also be different from the materials constituting the other plates (300, 304, 306).

[0058] In this example, the elastic layer (302) is formed as a liquid-impermeable flexible membrane. In some versions, the elastic layer (302) is gas-permeable, although liquid-impermeable. In some such versions, specific areas of the elastic layer (302) are treated to be gas-permeable, while untreated areas of the elastic layer (302) are gas-impermeable. As described below, the elastic layer (302) can be used to drive fluid through the processing chip (200) via a peristaltic pumping action. Furthermore, as described below, the elastic layer (302) can be used to provide valves at various locations along the processing chip (200). In some versions, a single sheet of elastic material spans the width of the processing chip (200) to form the elastic layer (302). In some other versions, two or more discrete sheets of elastic material are used to form the elastic layer (302), these discrete sheets of elastic material being positioned at different locations across the width of the processing chip (200). By way of example only, the elastic layer (302) may include a membrane comprising a polydimethylsiloxane (PDMS) elastomer film.

[0059] like Figures 4A-4FMost clearly visible, the first and second plates (300, 304) work together to define multiple chambers (320, 322, 324, 326), and the elastic layer (302) divides each chamber (320, 322, 324, 326) into corresponding upper chamber area (330) and lower chamber area (332). Figure 3 The rooms shown (224, 225, 230, 234, 250, 252, 260, 262, 264, 270) can be like... Figures 4A-4F The chambers (320, 322, 324, 326) shown are configured and operated in the same manner. For example, chamber (320) can be similar to chamber (264), chamber (322) can be similar to chamber (260), chamber (324) can be similar to chamber (262), and chamber (326) can be similar to chamber (250).

[0060] like Figures 4A-4F As shown, a fluid port (220) is formed through the first plate (300). A corresponding opening (342) is formed through the region below the fluid port (220) of the elastic layer (302). A fluid passage (222) extends from the opening (342) to the lower chamber region (332) of the first chamber (320). As described above, the fluid port (220) is configured to receive a fluid line (206) from the fluid interface assembly (109). The distal end of the fluid line (206) is configured to seal against a region of the elastic layer (302) exposed through the fluid port (220) and through the opening (342) for the transfer of fluid (207). In some versions, a spring or other elastic member provides an elastic bias to the fluid line (206), pushing the distal end of the fluid line (206) against the region of the elastic layer (302) exposed by the fluid port (220), thereby maintaining a seal. Fluid (207) from the fluid line (206) reaches the lower chamber region (332) of the first chamber (320) via the fluid channel (222). As described in more detail below, the fluid (207) can be further transferred from the first chamber (320) to other chambers (322, 324, 326) by peristaltic pumping action provided via the elastic layer (302). After reaching the fourth chamber (326), the fluid (207) can be further transferred to other chambers or other features in the processing chip (200), can be transferred to storage vials in the reagent storage frame (107), or can be processed in other ways. Therefore, the path of the fluid (207) does not necessarily terminate in the fourth chamber (326). It should also be understood that Figure 3 Any other fluid ports shown (221, 266) can be like Figures 4A-4F Configured and operated as shown in the fluid port (220).

[0061] A pressure port (240) is formed through the first plate (300). A corresponding opening (344) is formed through the region below the pressure port (240) of the elastic layer (302). A pressure passage (244) extends from the opening (344) to the upper chamber region (330) of the first chamber (320). As described above, the pressure port (240) is configured to receive a pressure line (208) from the fluid interface assembly (109), thereby receiving pressurized gas from a pressure source (117). The distal end of the pressure line (208) is configured to seal against a region of the elastic layer (302) exposed through the pressure port (240) and through the opening (344) for the delivery of either positive or negative pressurized gas. In some versions, a spring or other elastic member provides an elastic bias to the pressure line (208), pushing the distal end of the pressure line (208) against the region of the elastic layer (302) exposed by the pressure port (240), thereby maintaining a seal. Positive or negative pressurized gas from the pressure line (208) reaches the upper chamber area (330) of the fourth chamber (326) via the pressure channel (244).

[0062] Although Figures 4A-4F The diagram depicts only one pressure line (208) coupled to the processing chip (200), but the processing chip (200) may have several coupled pressure lines (208), where these pressure lines (208) independently apply positive or negative pressure to corresponding chambers (320, 322, 324, 326) of the processing chip (200). In some versions, one or more chambers (320, 322, 324, 326) have their own dedicated pressure lines (208) and corresponding pressure channels (244). Alternatively, one or more chambers (320, 322, 324, 326) may share a common pressure line (208) via the same pressure channel (244) or via separate pressure channels (244). Although Figures 4A-4F The pressure channel (244) is depicted as being formed through the second plate (304), but some pressure channels (244) (or regions of pressure channels (244)) may be formed by the first plate (300). For example, some pressure channels (244) (or regions of pressure channels (244)) may be formed between a recess in the lower surface of the first plate (300) and the top surface of the elastic layer (302).

[0063] IV. Examples of valve regulation and peristaltic pumping driven by an elastic layer As described above, the elastic layer (302) can be operated to: drive fluid through the processing chip (200) by a peristaltic pumping action; and to prevent fluid movement through the processing chip (200) by providing a valve-regulating action. An example of this operation is in the passage of fluid through the processing chip (200). Figures 4A-4FThe sequence is illustrated in the description. In this example, chambers (320, 324) are used as valve chambers, while chamber (322) is used as a metering chamber. Chamber (326) is used as a working chamber, allowing synthesis, purification, dialysis, compounding, concentration, or some other processing to be carried out in chamber (326). This configuration, arrangement, and use of chambers (320, 322, 324, 326) is provided as an illustrative example. Chambers (320, 322, 324, 326) may alternatively be configured, arranged, and used in other ways.

[0064] Figure 4A A processing chip (200) is shown in the following states: where fluid has not yet been delivered to the processing chip (200); and pressurized gas has not yet been delivered to the processing chip (200). Figure 4B In this configuration, positively pressurized gas is delivered to the upper chamber region (330) of chamber (324), negatively pressurized gas is delivered to the upper chamber region (330) of chambers (320, 322), and fluid (207) is delivered to chambers (320, 322). Under these conditions, the positively pressurized gas causes partial deformation of the elastic layer (302) in chamber (324), causing the elastic layer (302) to abut against the surface of the lower chamber region (332) of chamber (324). This abutment of the elastic layer (302) against the surface of the lower chamber region (332) of chamber (324) prevents fluid (207) from entering chamber (324), thus ensuring that chamber (324) remains within the chamber. Figure 4B In the indicated state, it operates like a closed valve. The negatively pressurized gas in the upper chamber region (330) of chambers (320, 322) causes the corresponding portion of the elastic layer (302) in chambers (320, 322) to deform and settle against the upper chamber region (330) of chambers (320, 322). This allows the fluid (207) to occupy the full volume of chambers (320, 322).

[0065] Upon reaching Figure 4B Following the state shown, pressurized gas is transferred to the upper chamber region (330) of chamber (320), while the aerodynamic state of chambers (322, 324) can remain unchanged. This results in Figure 4C The state is shown. As shown, the pressurized gas causes partial deformation of the elastic layer (302) in the chamber (320), causing the elastic layer (302) to abut against the surface of the lower chamber region (332) of the chamber (320). This abutment of the elastic layer (302) against the surface of the lower chamber region (332) of the chamber (320) drives the fluid (207) to flow out of the chamber (320), and causes the chamber (320) to... Figure 4C In the indicated state, it operates like a closed valve. However, in Figure 4CIn the illustrated state, the volume of the fluid (207) in the chamber (322) is unaffected. Therefore, the chamber (322) can be used to provide metering of the fluid (207) so that only a precise, predetermined volume of fluid (207) is further conveyed along the processing chip (200). As an example only, the order of magnitude of this metering volume could be approximately 10 nL, 20 nL, 25 nL, 50 nL, 75 nL, 100 nL, 1 μL, 5 μL, etc.

[0066] Once the appropriate metering volume has been reached, the negatively pressurized gas is transferred to the upper chamber region (330) of chambers (324, 326), while the aerodynamic state of chambers (320, 322) can remain unchanged. This results in Figure 4D The state is shown. As shown, the negatively pressurized gas in the upper chamber region (330) of chambers (324, 326) causes the corresponding portion of the elastic layer (302) in chambers (324, 326) to deform and abut against the surface of the upper chamber region (330) of chambers (324, 326). This effectively opens the valve formed by chamber (324) and places chamber (326) in a state of receiving fluid (207). This also creates a negative pressure in chamber (324) that draws fluid (207) from chamber (322) into chamber (324).

[0067] When the valve formed by chamber (324) is open, pressurized gas is delivered to the upper chamber region (330) of chamber (322), while the aerodynamic state of chambers (320, 324, 326) can remain unchanged. This results in Figure 4E The state is shown. As shown, the positively pressurized gas in the upper chamber region (330) of chamber (322) causes the corresponding portion of the elastic layer (302) in chamber (322) to deform and abut against the surface of the lower chamber region (332) of chamber (322). This deformation of the elastic layer (302) expels fluid (207) from chamber (322). Since the valve formed by chamber (320) is closed and the valve formed by chamber (324) is open, fluid (207) travels from chamber (322) into chamber (324). In this example, the capacity of chamber (322) is greater than the capacity of chamber (324), causing fluid (207) from chamber (322) to overflow from chamber (324) into chamber (326).

[0068] Once the fluid (207) has been transferred from chamber (322) to chambers (324, 326), pressurized gas is transferred to the upper chamber region (330) of chamber (324), while the aerodynamic state of chambers (320, 322, 326) can remain unchanged. This results in Figure 4FThe state is shown. As shown, the positively pressurized gas in the upper chamber region (330) of chamber (324) causes the corresponding portion of the elastic layer (302) in chamber (324) to deform and abut against the surface of the lower chamber region (332) of chamber (324). This deformation of the elastic layer (302) expels fluid (207) from chamber (324). Since the deformed portion of the elastic layer (302) in chamber (324) effectively seals chamber (324) from chamber (322) (e.g., causing chamber (324) to operate like a closed valve), fluid (207) travels from chamber (324) into chamber (326).

[0069] exist Figure 4F At the stage shown, fluid (207) has been emptied from chambers (320, 332, 324), and chamber (326) contains a precisely measured volume of fluid (207) in chamber (322). The fluid (207) in chamber (326) may be further processed within chamber (326) in accordance with the teachings herein. Alternatively, or as an alternative, the fluid (207) in chamber (326) may be transferred to one or more other chambers of the processing chip (200), may be transferred to vials in the reagent storage frame (107), or may be processed in other ways. Regardless of what is done to the fluid (207) after it has reached the chamber (326), it should be understood that the fluid (207) is sequentially transported along the chambers (320, 322, 324) to reach the chamber (326) via peristalsis, which is generated by the elastic layer (302) in response to the delivery of positively or negatively pressurized gas in a specific order to the upper chamber region (330) of the chambers (320, 322, 324, 326). This peristaltic pumping may be particularly advantageous for fluids that may be viscous or contain suspended particles (e.g., purified or captured beads). This peristaltic pumping via the selective deformation of the elastic layer (302) can also be referred to as pneumatic barrier deflection or "pneumatic deflection".

[0070] In some cases, it may be desirable to remove air or other gases from one or more fluid passages of the treatment chip (200). To achieve this, the treatment chip (200) may include one or more chambers configured to provide ventilation of the fluid passages or otherwise expel air from the fluid passages. For example, such ventilation or evacuation may be performed as part of an infusion process when fluid is initially introduced into the treatment chip (200). Alternatively, such ventilation or evacuation may be performed to release gases generated in the fluid during the formation of the therapeutic composition. Such a ventilation or gas release chamber may be referred to as a “vacuum cap”. In some versions, at least the area of ​​the elastic layer (302) located in the vacuum cap (if not the entire elastic layer (302)) is gas-permeable (while remaining liquid-impermeable). Negatively pressurized gas may be applied to the upper chamber region (330) of the chamber, which serves as the vacuum cap, and this negatively pressurized gas may draw air or gas from the fluid passages through the corresponding area of ​​the elastic layer (302). In some versions, the upper chamber region (330) of the chamber, which serves as a vacuum cap, includes one or more protrusions or stand-off features that prevent the corresponding area of ​​the elastic layer (302) from fully abutting the surface of the upper chamber region (330) of the chamber, which serves as a vacuum cap. This can further facilitate the evacuation of air or other gases via the vacuum cap.

[0071] V. Examples of processing chips with bead-containing chambers In some cases, it may be desirable to provide a version of the processing chip (200) that provides one or more processing chambers in which fluids can be processed with functionalized solid particles (e.g., purification beads); and to prevent these particles from obstructing the flow of fluid through the processing chip (200) and / or from traveling into areas of the processing chip (200) where the presence of these particles may be undesirable. For example, in a version of the processing chip (200) configured to provide RNA purification with one or more purification chambers, it may be desirable to provide purification beads in those purification chambers to facilitate the purification process. In this regard, it should be understood that various types of purification beads can be used to purify and / or isolate RNA (e.g., mRNA) from the IVT manufacturing process. In some cases, such beads may comprise a resin that effectively separates mRNA from components of the transcription reaction process (e.g., plasmid DNA, enzymes, and other IVT components) by selectively capturing mRNA via a polyadenylate (polyA) tail using a salt and water purification step. More specifically, such beads may comprise a rigid polymeric resin support matrix comprising cross-linked poly(styrene-divinylbenzene); and / or may have a polyhydroxyl surface coating. The surface of such beads can be functionalized with poly(dT). Each bead may have a predetermined particle size (e.g., diameter), for example, in the range of about 10 µm to about 200 µm, such as about 50 µm; and / or may each be porous to increase the surface area of ​​each bead, thereby effectively increasing the contact area between the bead and the fluid. An example of such beads is the POROS Oligo(dT) 25 affinity resin from Thermo Fisher Scientific of Waltham, Massachusetts. Other examples of particles that can be used for purification include urea particles, which may also each have a predetermined particle size (e.g., diameter), for example, in the range of about 10 µm to about 200 µm, such as about 50 µm.

[0072] Therefore, it may be desirable to provide a variant of the processing chip (200) having channels and valves, the size and configuration of which can guide such particles to at least one processing chamber, such as at least one purification chamber, and filter such particles from the processed fluid to prevent the particles from leaving the at least one processing chamber along with the processed fluid.

[0073] Furthermore, in some cases, when the elastic layer (302) aerodynamically deforms against the surface of the chambers (320, 322, 324, 326) to expel fluid from the chambers (320, 322, 324, 326), the elastic layer (302) may tend to trap fluid clumps against the bottom plate and / or sidewalls of the chambers (320, 322, 324, 326), such that not all fluid leaves the chambers (320, 322, 324, 326) when the elastic layer (302) aerodynamically deforms to expel fluid from the chambers (320, 322, 324, 326). Therefore, it may be desirable to provide a feature in the chamber that prevents the elastic layer from trapping fluid clumps within the chamber when it aerodynamically deforms to expel fluid from the chamber.

[0074] Figures 5 to 8 An example of a processing chip (400) providing at least some (if not all) of the features and functions described above is shown. Except as further described below, the processing chip (400) is similar to the processing chip (200) described above. In this respect, in conjunction with the rest of the system (100), the processing chip (400) can be used to provide in vitro synthesis, purification, concentration, formulation (e.g., encapsulating the therapeutic composition in a delivery carrier by a mixing process), and / or analysis of therapeutic compositions and / or templates, including but not limited to therapeutic polynucleotide templates, therapeutic polynucleotides, and / or therapeutic polynucleotide compositions. Alternatively, the processing chip (400) can be used in other processes.

[0075] like Figure 5 and Figure 7 As shown, the processing chip (400) of this example includes multiple fluid ports (410) and multiple pressure ports (420). Each fluid port (410) is configured to receive fluid from a corresponding fluid line (206) of the fluid interface assembly (109). Each pressure port (420) is configured to receive pressurized gas from a corresponding pressure line (208) of the fluid interface assembly (109). Also as Figure 5 and Figure 7As shown, this example includes a pair of purification bead ports (410a, 410b), each configured to receive purification beads or similar particles from and / or deliver beads or similar particles to a corresponding fluid line (206). For example, a first purification bead port (410a) may be configured to receive a slurry containing beads suspended in a liquid solution (e.g., 0.1M sodium chloride) from a first fluid line (206), and a second purification bead port (410b) may be configured to subsequently deliver the slurry to a second fluid line (206), as discussed in more detail below. In some cases, it may be desirable to permanently retain the beads within the processing chip (400). For example, one or both purification bead ports (410a, 410b) may be used only during the initial fabrication or initial setup of the processing chip (400) to introduce beads into the processing chip (400).

[0076] like Figure 7 As shown, the processing chip (400) of this example also includes multiple processing chambers (450, 460, 470a, 470b) that can be used for different purposes in the production of therapeutic compositions as described herein. By way of example only, the processing chambers (450, 460, 470a, 470b) can be used to provide, purify, dialyze, compound, and concentrate one or more therapeutic compositions; or to perform any other suitable function. In this example, processing chamber (450) is configured for IVT reactions and may therefore also be referred to as a “reaction chamber”; processing chamber (460) is configured for DNA digestion and may therefore also be referred to as a “digestion chamber”; and processing chambers (470a, 470b) are configured for RNA purification and may therefore also be referred to as “purification chambers”. The processing chip (400) may have any other suitable number and / or type of processing chambers (450, 460, 470a, 470b). Figure 8 As shown, the processing chip (400) in this example also includes a first layer (500), a second layer (600), a third layer (700), a fourth layer (800), a fifth layer (900), a sixth layer (1000), and a seventh layer (1100). Each of these layers (500, 600, 700, 800, 900, 1000, 1100) and their structural and functional relationships with each other will be described in more detail below.

[0077] In this example, both the fluid port (410) and the pressure port (420) are located on the top region of the processing chip (400), at least in Figure 5Within the reference frame, this is the case. In some other versions, the fluid ports (410) are located on the top region of the processing chip (400), while the pressure ports (420) are located on the bottom region of the processing chip (400). In some other versions, the fluid ports (410) are located on the bottom region of the processing chip (400), while the pressure ports (420) are located on the top region of the processing chip (400). As yet another variation, some fluid ports (410) may be located on the top region of the processing chip (400), while others are located on the bottom region of the processing chip (400). Similarly, some pressure ports (420) may be located on the top region of the processing chip (400), while others are located on the bottom region of the processing chip (400).

[0078] Figures 9 to 11 The first layer (500) is shown in more detail. In this example, the first layer (500) is in the form of a rigid plate. In some (but not all) versions, the first layer (500) may be substantially translucent to visible and / or ultraviolet light. “Substantially translucent” means that at least 90% (in some cases, 100%) of light is transmitted through the material compared to a translucent material. The first layer (500) may include glass, plastic, silicone, and / or any other suitable material. In some versions, the first layer (500) is formed as a laminate of two or more material layers, such that the first layer (500) does not necessarily need to be formed as a single, homogeneous material continuum.

[0079] like Figures 9 to 11 As shown, the first layer (500) includes an upper surface (502), a lower surface (504), a first plurality of openings (510) formed through the two surfaces (502, 504), and a second plurality of openings (520) formed through the two surfaces (502, 504). The openings (510) form part of fluid ports (410), such that each fluid port (410) has a corresponding opening (510), and the openings (510) are configured to receive fluid lines (206). The openings (520) form part of pressure ports (420), such that each pressure port (420) has a corresponding opening (520), and the openings (520) are configured to receive pressure lines (208). A plurality of processing chamber portions (552) are formed as recesses in the lower surface (504), collectively providing a protruding area (550) in the upper surface (502). Each processing chamber section (552) forms a “dry” area as described herein for the corresponding processing chambers (450, 460, 470a, 470b).

[0080] In the illustrated example, the plurality of openings (510) include a pair of purification bead openings (510a, 510b) configured to allow purification beads (e.g., together with a liquid solution in which the beads can be suspended) to pass through the first layer (500). In this example, the first purification bead opening (510a) forms part of a first purification bead port (510a), and the second purification bead opening (510b) forms part of a second purification bead port (510b), such that each purification bead port (410a, 410b) has a corresponding opening (510a, 510b).

[0081] like Figures 10-11 Ideally visible, the lower surface (504) of the first layer (500) further defines a plurality of pneumatic channels (522), which are formed as recesses in the lower surface (504). Each pneumatic channel (522) is pneumatically connected to a corresponding opening (520). Each pneumatic channel (522) is also configured to define a space between the first layer (500) and the second layer (600) so that pressurized gas can be conveyed along the pneumatic channel (522).

[0082] The lower surface (504) of the first layer (500) further defines a plurality of valve chamber portions (530) and a plurality of pump chamber portions (540). The valve chamber portions (530) and pump chamber portions (540) are formed as recesses in the lower surface (504) and are pneumatically connected to corresponding pneumatic channels (522). Therefore, pressurized gas can be transmitted from the pressure port (420) to the valve chamber portions (530) and pump chamber portions (540) via the corresponding pneumatic channels (522).

[0083] The valve chamber portion (530) forms the “dry” region of the valve chamber as described herein; while the pump chamber portion (540) forms the “dry” region of the pump chamber as described herein. In this example, the valve chamber portion (530) has a circular shape, while some pump chamber portions (540) have an oblong or stadium shape, and other pump chamber portions (540) have a circular shape, but other shapes may be used. By way of further example only, the valve chamber associated with the valve chamber portion (530) can operate as described in the chambers (320, 322) above; while the pump chamber associated with the pump chamber portion (540) can operate as described in the chambers (322, 326) above. Although each chamber portion (530, 540) in this example has its own associated pneumatic passage (522) and pressure port (420), each processing chamber portion (552) in this example also has its own associated pneumatic passage (522) and pressure port (420). Therefore, each chamber section (530, 540, 552) can be pneumatically pressurized independently of the other chamber sections (530, 540, 552).

[0084] The second layer (600) lies between the first layer (500) and the third layer (700). In this example, the second layer (600) is in the form of first and second liquid-impermeable flexible membranes (600a, 600b) arranged vertically stacked and laminated together. As described below, the second layer (600) can be used to drive fluid through the processing chip (400) via a peristaltic pumping action. Furthermore, as described below, the second layer (600) can be used to provide valves at various locations along the processing chip (400). By way of example only, one or both flexible membranes (600a, 600b) of the second layer (600) may comprise polydimethylsiloxane (PDMS) elastomer membranes. Alternatively, any other suitable one or more materials may be used to form the second layer (600). Although the first flexible membrane (600a) is shown disposed below the second flexible membrane (600b), such that the first flexible membrane (600a) may be referred to as the lower flexible membrane (600a) and the second flexible membrane (600b) may be referred to as the upper flexible membrane (600b), the first flexible membrane (600a) may alternatively be disposed above the second flexible membrane (600b).

[0085] The second layer (600) can be constructed and operated in the same manner as the elastic layer (302) described above. Therefore, the second layer (600) can pass through the chambers defined by the first and third layers (500, 700) to divide these chambers into an upper chamber region (which receives pneumatic pressure) and a lower chamber region (which receives fluid). Such chambers include processing chambers (450, 460, 470a, 470b), valve chambers as described herein, and pump chambers as described herein. Therefore, the second layer (600) can be further pneumatically deformed to provide valve regulation, peristaltic pumping, etc., within the processing chip (400).

[0086] like Figure 12 and Figure 13As shown, the second layer (600) includes a plurality of openings (602a, 602b) formed through the flexible membranes (600a, 600b). The openings (602a, 602b) may be positioned below the corresponding openings (510, 520) at the fluid port (410) and / or the pressure port (420) to allow fluid and / or pressurized gas to be conveyed through the second layer (600) at the fluid port (410) and / or the pressure port (420), respectively. For example, as described above, each fluid port (410) is configured to receive a fluid line (206) from the fluid interface assembly (109). The distal end of the fluid line (206) may be configured to seal against the area of ​​the second layer (600) exposed by the fluid port (410) and to deliver fluid through the corresponding openings (602a, 602b) in the second layer (600). In some versions, a spring or other elastic member provides an elastic bias to the fluid line (206), pushing the distal end of the fluid line (206) against the area of ​​the second layer (600) exposed by the fluid port (410), thereby maintaining a seal. Fluid from the fluid line (206) can then be further transferred via the processing chip (400), as described in more detail below.

[0087] like Figure 12 As shown, the second flexible membrane (600b) also includes a plurality of holes (e.g., slits) (604), each hole being configured to be vertically aligned with a corresponding processing chamber (450, 460, 470a, 470b), valve chamber, or pump chamber. For example, each hole (604) may be located directly below a corresponding upper chamber region and / or directly above a corresponding lower chamber region. Each hole (604) of the second flexible membrane (600b) may be located directly above or below a corresponding portion of the first flexible membrane (600a), which is configured to deflect within the corresponding processing chamber (450, 460, 470a, 470b), valve chamber, or pump chamber. Therefore, the hole (604) can accommodate the deflection of the first flexible membrane (600a) by preventing the second flexible membrane (600b) of the second layer (600) from forming wrinkles or otherwise interfering with the deflection of the first flexible membrane (600a) of the second layer (600) in the processing chamber (450, 460, 470a, 470b), valve chamber or pump chamber.

[0088] As a further example only, the second layer (600) may be configured and operated at least in part according to the teachings of U.S. Patent Application No. 63 / 453,206, filed March 20, 2023, entitled “Microfluidic Apparatus with Elastic Layers and Contoured Surface,” the disclosure of which is incorporated herein by reference in its entirety.

[0089] In this example, the region of each chamber in the processing chip (400) above the second layer (600) constitutes a "dry" chamber region because this region receives pressurized gas to pneumatically deflect the second layer (600). Conversely, the region of each chamber in the processing chip (400) below the second layer (600) constitutes a "wet" chamber region because this region receives fluid. However, these "wet" and "dry" roles can be reversed in some variations. In other words, some variations of the processing chip (400) may supply pressurized gas to the lower region of each chamber, such that the lower chamber region constitutes a "dry" chamber region; while the upper chamber region receives fluid, such that the upper chamber constitutes a "wet" chamber region.

[0090] Figures 14 to 18 The third layer (700) is shown in more detail. In this example, the third layer (700) is in the form of a rigid plate. In some (but not necessarily all) versions, the third layer (700) is substantially translucent to visible and / or ultraviolet light. The third layer (700) may contain glass, plastic, silicone, and / or any other suitable material. In some versions, the third layer (700) is formed as a laminate of two or more material layers, such that the third layer (700) does not necessarily need to be formed as a single, homogeneous material continuum.

[0091] like Figures 12 to 14 As shown, the third layer (700) includes an upper surface (702), a lower surface (704), and a plurality of openings (710) formed through the two surfaces (702, 704). A plurality of processing chamber portions (752) are formed as recesses in the upper surface (702), collectively providing a protruding area (750) in the lower surface (704). Each processing chamber portion (752) forms a “wet” area of ​​a corresponding processing chamber (450, 460, 470a, 470b) as described herein. Some of the openings (710) form part of a pressure port (420), such that each pressure port (420) has a corresponding opening (710), and such that the openings (710) are configured to allow pressurized gas to pass through the third layer (700). As described above, the corresponding openings (602a, 602b) can be formed through the second layer (600) such that at least some of the openings (602a, 602b) in the second layer (600) are located between at least some of the openings (520) in the first layer (500) and the openings (710) in the third layer (700).

[0092] In the example shown, the plurality of openings (710) includes a plurality of purification bead openings (710a, 710b, 710c, 710d) configured to allow purification beads (e.g., together with a liquid solution in which the beads can be suspended) to pass through the third layer (700). In this example, the first purification bead opening (710a) forms part of a first purification bead port (410a), while the fourth purification bead opening (710d) forms part of a second purification bead port (410b), such that each purification bead port (410a, 410b) has a corresponding opening (710a, 710d).

[0093] Each processing chamber section (752) has a sidewall (753) extending downward from the upper surface (702) toward the corresponding base plate (755). For example... Figure 14 and Figure 16 As shown, the upper surface (702) of the third layer (700) further defines a plurality of fluid channels (712), which are formed as recesses in the upper surface (702). Each fluid channel (712) is in fluid communication with a corresponding opening (710). Each fluid channel (712) is also configured to define a space between the third layer (700) and the second layer (600) so that fluid can be transported along the fluid channel (712).

[0094] In the illustrated example, the plurality of fluid channels (712) include a plurality of purification bead channels (712a, 712b, 712c, 712d, 712e) configured to provide a transport path for purification beads (e.g., along with a liquid solution in which the beads may be suspended) to and from a “wet” area of ​​the first purification chamber (470a). In this regard, the purification bead channels (712a, 712b, 712c, 712d, 712e) may each be sized and configured to accommodate beads of a predetermined particle size (e.g., diameter) flowing through them, while mitigating any risk of these beads clogging or otherwise obstructing the flow of fluid. For this purpose, the respective cross-sectional dimensions (e.g., width, depth, diameter, etc.) of the purification bead channels (712a, 712b, 712c, 712d, 712e) may be significantly larger than the corresponding cross-sectional dimensions of some or all other fluid channels (712) formed in the upper surface (702). In some cases, the purification bead channels (712a, 712b, 712c, 712d, 712e) may each have a minimum cross-sectional size that is at least five times larger than the predetermined particle size of each bead. With each bead having a predetermined particle size of approximately 50 µm, the minimum cross-sectional size of each purification bead channel (712a, 712b, 712c, 712d, 712e) may be at least approximately 250 µm. Each purification bead channel (712a, 712b, 712c, 712d, 712e) may have a width ranging from about 200µm to about 800µm, for example, a width ranging from about 200µm to about 750µm, and may have a depth ranging from about 200µm to about 750µm (e.g., relative to the upper surface (702)); while some or all of the other fluid channels (712) formed in the upper surface (702) may have a width ranging from about 100µm to about 400µm, and may have a depth ranging from about 100µm to about 500µm (e.g., relative to the upper surface (702)). For example, each purification bead channel (712a, 712b, 712c, 712d, 712e) may have a width of about 500 µm and a depth of about 250 µm (e.g., relative to the upper surface (702)); while some or all of the other fluid channels (712) formed in the upper surface (702) may have a width of about 200 µm and a depth of about 100 µm (e.g., relative to the upper surface (702)). As another example, each purification bead channel (712a, 712b, 712c, 712d, 712e) may have a width of about 500 µm and a depth of about 500 µm. As yet another example, each purification bead channel (712a, 712b, 712c, 712d, 712e) may have a width of about 800 µm and a depth of about 400 µm.Alternatively, each purification bead channel (712a, 712b, 712c, 712d, 712e) may have any other suitable cross-sectional size selected to accommodate beads of any predetermined particle size.

[0095] The upper surface (702) of the third layer (700) further defines a plurality of valve chamber portions (730) and a plurality of pump chamber portions (740). The valve chamber portions (730) and pump chamber portions (740) are formed as recesses in the upper surface (702) and are in fluid communication with corresponding fluid channels (712). Thus, fluid can be transferred from the fluid port (410) to the valve chamber portions (730) and pump chamber portions (740) via the corresponding openings (710) and fluid channels (712).

[0096] The valve chamber portion (730) forms the “wet” region of the valve chamber as described herein (opposite to the corresponding valve chamber portion (530)); while the pump chamber portion (740) forms the “wet” region of the pump chamber (opposite to the corresponding pump chamber portion (540)) as described herein. In this example, the valve chamber portion (730) has a circular shape, while some pump chamber portions (740) have an oblong or stadium shape, and other pump chamber portions (740) have a circular shape, but other shapes may be used. By way of further example only, the valve chamber associated with the valve chamber portion (730) may operate as described above in the chambers (320, 322); while the pump chamber associated with the pump chamber portion (740) may operate as described above in the chambers (322, 326). In this example, each chamber portion (730, 740) has its own associated fluid passage (712) and opening (710). Therefore, each chamber section (730, 740) can receive fluid independently of the other chamber sections (730, 740).

[0097] Each processing chamber section (752) also has at least one corresponding fluid passage (712) leading to it, such that fluid can be transferred to / from each processing chamber (450, 460, 470a, 470b) through the fluid passage (712) leading to the processing chamber section (752) associated with the processing chamber (450, 460, 470a, 470b). Also in this example, a pump chamber section (740) and a set of valve chamber sections (730) are located between an opening (710) and each fluid passage (712) leading to each corresponding processing chamber section (752). The pump and valves defined by these sections (740, 730) can thus drive and regulate the transfer of fluid to / from each processing chamber (450, 460, 470a, 470b).

[0098] In the illustrated example, purification bead channels (712c, 712d) lead to a processing chamber portion (752) associated with the first purification chamber (470a), allowing purification beads to be conveyed to / from the first purification chamber (470a) via the purification bead channels (712c, 712d). For example, purification bead channel (712c) may define a purification bead inlet channel (also referred to as a "particle inlet channel") leading to the first purification chamber (470a) for conveying purification beads to the first purification chamber (470a), and purification bead channel (712d) may define a purification bead outlet channel (also referred to as a "particle outlet channel") extending from the first purification chamber (470a) for conveying purification beads from the first purification chamber (470a).

[0099] In this example as well, the plurality of valve chamber portions (730) include a plurality of purification bead valve chamber portions (730a, 730b), and the plurality of pump chamber portions (740) include a purification bead pump chamber portion (740a), each inserted between a corresponding opening (710) and a corresponding one of the purification bead channels (712c, 712d) leading to a processing chamber portion (752) associated with the first purification chamber (470a). Thus, the pump and valve defined by these portions (740a, 730a, 730b) can drive and regulate the delivery of purification beads to / from the first purification chamber (470a). In this respect, each purification bead valve chamber portion (730a, 730b) is arranged opposite to the corresponding purification bead valve chamber portion (530a, 530b) of the plurality of valve chamber portions (530), and the purification bead pump chamber portion (740a) is arranged opposite to the purification bead pump chamber portion (540a) of the plurality of pump chamber portions (540).

[0100] The valves defined by the purified bead valve chamber portions (530a, 530b, 730a, 730b) and the pumps defined by the purified bead pump chamber portions (540a, 740a) can each be sized and configured to accommodate beads (e.g., together with a liquid solution in which the beads can be suspended) flowing through them, while mitigating any risk of such beads clogging or otherwise obstructing fluid flow. For example, the purified bead valve chamber portions (530a, 530b, 730a, 730b) can each have a cross-sectional dimension (e.g., diameter) larger than some or all of the other valve chamber portions (530, 730), such that the valves defined by the purified bead valve chamber portions (530a, 530b, 730a, 730b) can be configured to accommodate a larger fluid volume than the valves defined by some or all of the other valve chamber portions (530, 730). In some cases, the purification bead valve chamber portion (530a, 530b, 730a, 730b) may each have a diameter of about 500µm to about 6 mm, for example, about 2.1 mm.

[0101] As shown in the figure, the purification bead pump chamber portions (540a, 740a) of this example each have a circular shape, rather than an elongated oval or stadium shape like some or all of the other pump chamber portions (540, 740), such that the shape and / or size of each purification bead pump chamber portion (540a, 740a) is similar to that of the valve chamber portion (530, 730). Alternatively or additionally, the purification bead pump chamber portions (540a, 740a) may each have a depth greater than some or all other pump chamber portions (540, 740) (e.g., relative to the corresponding upper surface (502, 702)). In some cases, the relatively large depth of the purification bead pump chamber portions (540a, 740a) may allow the pump defined by the purification bead pump chamber portions (540a, 740a) to accommodate a larger fluid volume than the pump defined by some or all of the other pump chamber portions (540, 740). In some other cases, the relatively large depth of the purification bead pump chamber portions (540a, 740a) combined with their relatively small cross-sectional dimensions (e.g., because each of the purification bead pump chamber portions (540a, 740a) has a circular shape rather than an oblong or stadium shape) can result in the pump defined by the purification bead pump chamber portions (540a, 740a) being configured to contain substantially the same fluid volume as the pump defined by some or all of the other pump chamber portions (540, 740). For example, the pump defined by the purification bead pump chamber portions (540a, 740a) and the pump defined by some or all of the other pump chamber portions (540, 740) can each be configured to contain a fluid volume of about 50 nL to about 140 µL, such as about 3 µL of fluid. In this case, the relatively large depth of the purification bead pump chamber section (540a, 740a) can still help mitigate any risk of the beads becoming clogged in the pump defined by the purification bead pump chamber section (540a, 740a).

[0102] For example Figure 14 and Figure 16As seen, the third layer (700) includes multiple bridging channels (760), sidewall channels (754), and bottom plate channels (756). The sidewall channels (754) can also be referred to as "straws". Each bridging channel (760) is formed as a recess in the upper surface (702); each bottom plate channel (756) is formed as a recess in the corresponding bottom plate (755); and each sidewall channel (754) is formed as a recess in the corresponding sidewall (753), which extends upward from the corresponding bottom plate channel (756) toward the upper surface (702). When the second layer (600) deforms downward toward the corresponding base plate (755), the presence of the sidewall channels (754) and / or the base plate channels (756) can provide a path for fluid to escape from the corresponding processing chambers (450, 460, 470a, 470b), such that the sidewall channels (754) and / or the base plate channels (756) can prevent fluid clumps from being trapped in local areas of the deformed second layer (600) between the corresponding base plate (755) and / or the sidewall (753) (or otherwise reduce the risk of forming such clumps).

[0103] As shown, at least some sidewall channels (754) extend from corresponding base plate channels (756) to corresponding bridging channels (760), such that each bridging channel (760) is flanked by a pair of adjacent sidewall channels (754). Each bridging channel (760) and the pair of adjacent sidewall channels (754) are configured to provide a path for transferring fluid from one processing chamber (450, 460, 470a, 470b) to an adjacent processing chamber (450, 460, 470a, 470b).

[0104] In the example shown, multiple bridging channels (760) include a pair of purification bridging channels (760a, 760b) extending between processing chamber portions (752) associated with purification chambers (470a, 470b). In some versions, the purification bridging channels (760a, 760b) are configured to provide a path for conveying purification beads (e.g., together with a liquid solution in which the beads can be suspended) between the purification chambers (470a, 470b). In this regard, the purification bridging channels (760a, 760b) can be individually sized and configured to allow beads to flow through them while reducing any risk of these beads clogging or otherwise obstructing the flow of fluid. For example, each purification bridging channel (760a, 760b) may have a width of about 250µm to about 750µm, such as about 500µm, and a depth of about 250µm to about 750µm (e.g., relative to the upper surface (702)), such as about 500µm. Alternatively, each purification bridging channel (760a, 760b) may have any other suitable cross-sectional size selected to accommodate beads of any predetermined particle size.

[0105] like Figure 15 As shown, the third layer (700) also includes a plurality of fluid channels (714) on its lower surface (704). The fluid channels (714) are formed as recesses on the lower surface (704). Each fluid channel (714) is also configured to define a space between the third layer (700) and the fourth layer (800) so that fluid can be transported along the fluid channel (714). An opening (710) is located at each end of each fluid channel (714). Thus, the opening (710) and the fluid channel (714) cooperate to allow fluid to be transported from one area on the upper surface (702) of the third layer (700) to another area on the upper surface (702) of the third layer (700) via the lower surface (704) of the third layer (700), which makes fluid transport easier compared to transporting fluid only through the fluid channels (712) in the upper surface (702). In other words, including a fluid channel (714) on the lower surface (704) allows fluid to be transported along a path, and if that path is moved from the lower surface (704) to the upper surface (702), the path would originally intersect with the fluid path provided via the fluid channel (712).

[0106] In the example shown, the multiple fluid channels (714) include multiple purification bead channels (714a, 714b) which are sized and configured similarly to the purification bead channels (712a, 712b, 712c, 712d, 712e) to allow beads of a predetermined particle size (e.g., diameter) to flow through them while reducing any risk of these beads clogging or otherwise obstructing the flow of fluid. The first purification bead channel (714a) of this example extends from the first purification bead opening (710a) to the second purification bead opening (710b) (which communicates with the first purification bead channel (712a) on the upper surface (702)), while the second purification bead channel (714b) of this example extends from the fourth purification bead opening (710d) to the third purification bead opening (710c) (which communicates with the fifth purification bead channel (712e) on the upper surface (702)).

[0107] Now refer to Figure 17 and Figure 18 And continue to refer to Figure 14 and Figure 16The third layer (700) of this example also includes a plurality of purification bead filters (770a, 770b, 770c) on the upper surface (702). The purification bead filters (770a, 770b, 770c) may also be referred to as a “fritting structure”. Each of the purification bead filters (770a, 770b, 770c) is inserted between a corresponding processing chamber section (752) associated with the purification chamber (470a, 470b) and one or more corresponding fluid channels (712) (e.g., one or more corresponding fluid outlet channels). More specifically, a first purification bead filter (770a) is inserted between a processing chamber portion (752) associated with the first purification chamber (470a) and a plurality of corresponding fluid channels (712), the fluid channels (712) defining fluid outlet channels for conveying fluid output from the first purification chamber (470a); a second purification bead filter (770b) is inserted between a processing chamber portion (752) associated with the second purification chamber (470b) and a corresponding fluid channel (712), the fluid channels (712) defining waste outlet channels for conveying fluid waste from the second purification chamber (470b); and a third purification bead filter (770c) is inserted between a processing chamber portion (752) associated with the second purification chamber (470b) and a corresponding fluid channel (712), the fluid channels (712) defining UV sampling outlet channels for conveying fluid samples from the second purification chamber (470b). The purification bead filters (770a, 770b, 770c) are each configured to provide a path for conveying fluid from the respective purification chamber (470a, 470b) to one or more respective fluid channels (712), while inhibiting the flow of beads from the respective purification chamber (470a, 470b) to one or more respective fluid channels (712), thereby retaining such beads within the purification chamber (470a, 470b).

[0108] In this respect, each of the purification bead filters (770a, 770b, 770c) includes an array of filter channels (772) formed as recesses in the upper surface (702), a filter inlet (774) formed as recesses in the respective sidewalls (753) and extending between the inner ends of the respective filter channels (772), and a filter outlet (776) formed as recesses in the upper surface (702) and extending between the outer ends of the respective filter channels (772), such that each filter channel (772) extends between the respective filter inlet (774) and filter outlet (776). In the illustrated example, the filter inlet (774) of each purification bead filter (770a, 770b, 770c) extends along the outer peripheral edge of the corresponding processing chamber portion (752); the filter channels (772) of each purification bead filter (770a, 770b, 770c) extend substantially parallel to each other and substantially perpendicular to the corresponding filter inlet (774); and the filter outlet (776) of each purification bead filter (770a, 770b, 770c) extends substantially perpendicular to the corresponding filter channel (772) and substantially parallel to the corresponding filter inlet (774). Each filter channel (772) is also configured to define a space between a third layer (700) and a second layer (600), and each filter outlet (776) is similarly configured to define a space between the third layer (700) and the second layer (600), such that fluid can be conveyed along the filter channels (772) and the filter outlets (776) to one or more corresponding fluid channels (712).

[0109] Each filter channel (772) can be sized and configured to allow fluid to flow through it while preventing beads with a predetermined particle size (e.g., diameter) from flowing through it. In other words, the filter channels (772) of each purification bead filter (770a, 770b, 770c) can be configured to cooperate with each other to act as a sieve or "frit" to filter such beads from the fluid flowing through it. For this purpose, each filter channel (772) can have a cross-sectional dimension (e.g., width, depth, diameter, etc.) that is substantially smaller than the corresponding cross-sectional dimension of some or all of the fluid channels (712) formed in the upper surface (702). For example, each filter channel (772) can be relatively shallow compared to at least some or all of the fluid channels (712) formed in the upper surface (702). In some cases, each filter channel (772) can have at least one cross-sectional dimension that is substantially smaller than the predetermined particle size of each bead. With each bead having a predetermined particle size of about 50 µm, at least one cross-sectional dimension of each filter channel (772) can be substantially smaller than about 50 µm. For example, each filter channel (772) can have a depth in the range of about 10 µm to about 30 µm (e.g., relative to the upper surface (702)), such as a depth of about 25 µm. As another example, each filter channel (772) can have a depth in the range of about 10 µm to about 23 µm. Since one cross-sectional dimension of each filter channel (772) (e.g., its depth) can be small enough to prevent beads from flowing through it, another cross-sectional dimension of each filter channel (772) (e.g., its width) can in some cases be substantially equal to or even greater than the predetermined particle size of each bead. With each bead having a predetermined particle size of about 50 µm, another cross-sectional dimension of each filter channel (772) can be substantially equal to or greater than about 50 µm. For example, each filter channel (772) may have a width in the range of about 100µm to about 400µm, such as about 200µm. Alternatively, each filter channel (772) may have any other suitable cross-sectional dimensions, such as any other suitable depth, selected to filter out beads of any predetermined particle size.

[0110] The presence of a filter inlet (774) in each purification bead filter (770a, 770b, 770c) helps mitigate the risk of beads obstructing fluid flow to the corresponding filter channel (772) by providing space for the beads to aggregate without covering the corresponding filter channel (772). Alternatively, the presence of multiple filter channels (772) in each purification bead filter (770a, 770b, 770c) can reduce the impact on fluid flow from any blockage that may occur in a particular filter channel (772) by providing one or more alternative paths for the fluid to reach the corresponding filter outlet (776) (e.g., via one or more other unobstructed filter channels (772)). It should be understood that each purification bead filter (770a, 770b, 770c) may include any suitable number of filter channels (772).

[0111] As shown, at least some sidewall channels (754) extend from the base plate channel (756) of the processing chamber section (752) associated with one of the purification chambers (470a, 470b) to the corresponding purification bead filters (770a, 770b, 770c). Such sidewall channels (754) can be configured to provide a path for conveying purification beads (e.g., together with a liquid solution in which the beads can be suspended) from the respective purification chamber (470a, 470b) to the respective purification bead filters (770a, 770b, 770c). In this respect, such sidewall channels (754) can each be sized and configured to allow beads to flow through them while mitigating any risk of such beads clogging them or otherwise obstructing fluid flow. For example, each such sidewall channel (754) may have a width ranging from about 200 µm to about 800 µm, such as about 500 µm, and a depth ranging from about 100 µm to about 300 µm (e.g., relative to the corresponding sidewall (753)), such as about 100 µm. Alternatively, each sidewall channel (754) may have any other suitable cross-sectional dimensions.

[0112] In the illustrated example, the first purification bead filter (770a) is equipped with multiple (e.g., three) such sidewall channels (754), while the second and third purification bead filters (770b, 770c) are each equipped with a single such sidewall channel (754). The presence of multiple sidewall channels (754) leading to the first purification bead filter (770a) can help mitigate any risk of clogging of the beads therein by providing multiple paths for such beads to reach the first purification bead filter (770a). Alternatively or additionally, the presence of multiple sidewall channels (754) leading to the first purification bead filter (770a) can reduce the impact of any clogging that may occur within a particular sidewall channel (754) on fluid flow by providing one or more alternative paths for fluid to reach the first purification bead filter (770a) (e.g., via one or more other unclogging sidewall channels (754)). It should be understood that the purification bead filters (770a, 770b, 770c) can each be equipped with any other suitable number of sidewall channels (754).

[0113] In the example shown, another sidewall channel (754) extends from the bottom plate channel (756) of the processing chamber section (752) associated with the first purification chamber (470a) to the purification bead outlet channel (712d). This sidewall channel (754) can be configured to provide a path for conveying fluid from the first purification chamber (470a) to the purification bead outlet channel (712d).

[0114] exist Figure 19 An example of a flow path of particles through a portion of a processing chip (400) is shown. Such particles may, for example, comprise purification beads, such as POROSOligo(dT)25 affinity resin from Thermo Fisher Scientific in Waltham, Massachusetts. The purification beads may each have a predetermined particle size (e.g., diameter), for example, from about 10 µm to about 200 µm, such as about 50 µm, and / or may be suspended in a liquid solution (e.g., 0.1 M sodium chloride) to form a slurry. It should be understood that any other suitable purification beads or other types of functionalized particles (e.g., urea particles) may follow a similar flow path. Figure 19 The flow path is shown in the figure.

[0115] As indicated by arrow (A1), the purification bead can initially be delivered from the first purification bead port (410a) (e.g., in cooperation with the first fluid line (206)) through the first purification bead opening (710a), along the first purification bead channel (714a) on the lower surface (704), through the second purification bead opening (710b), along the first purification bead channel (712a) on the upper surface (702), through the purification bead pump chamber portion (740a), along the second purification bead channel (712b) on the upper surface (702), through the first purification bead valve chamber portion (730a), and along the third purification bead channel (712c) on the upper surface (702) into the "wet" area of ​​the first purification chamber (470a). The purification bead can process fluids delivered simultaneously, previously delivered fluids, or fluids subsequently delivered to the "wet" area of ​​the first purification chamber (470a).

[0116] As indicated by arrow (A2), the purification beads can then be transferred from the “wet” region of the first purification chamber (470a) to one or both purification bridging channels (760a, 760b) (e.g., via corresponding sidewall channels (754)) and into the “wet” region of the second purification chamber (470b). The purification beads can process simultaneously delivered fluid, previously delivered fluid, or fluid subsequently delivered to the “wet” region of the second purification chamber (470b). Also as indicated by arrow (A2), the purification beads can be transferred back and forth between the “wet” regions of the first and second purification chambers (470a, 470b). In some cases, the purification beads can be transferred back and forth between the “wet” regions of the first and second purification chambers (470a) and the second purification chamber (470b) along with the fluid being processed.

[0117] As indicated by arrow (A3), the purification bead can then be transferred from the “wet” area of ​​the first purification chamber (470a) to the fourth purification bead channel (712d) on the upper surface (702) (e.g., via a corresponding sidewall channel (754)), through the second purification bead valve chamber portion (730b), along the fifth purification bead channel (712e) on the upper surface (702), through the third purification bead opening (710c), along the second purification bead channel (714b) on the lower surface (704), through the fourth purification bead opening (710d), and into the second purification bead port (410b). The purification bead can be removed from the processing chip (400) via the second purification bead port (410b) (e.g., in cooperation with the second fluid line (206)), and / or can be returned to the first purification bead port (410a) for transfer back to the “wet” area of ​​the first purification chamber (470a). It should be understood that the second layer (600) can be pneumatically deformed in a manner similar to that discussed above to provide appropriate valved and / or peristaltic pumping for delivering the purification beads along each section of the flow path indicated by arrows (A1, A2, A3).

[0118] Because purification bead filters (770a, 770b, 770c) are present at each fluid outlet of the purification chambers (470a, 470b), the purification beads can be substantially confined within the flow paths indicated by arrows (A1, A2, A3), except for the purification bridging channels (760a, 760b) and the purification bead outlet channel (712d). Therefore, any purification beads that may be suspended in the fluid conveyed from the first purification chamber (470a) through the first purification bead filter (770a) can be filtered out of such fluid and remain in the first purification chamber (470a); such purification beads can then be removed from the first purification chamber (470a) via the purification bead outlet channel (712d). Similarly, any purification beads that may be suspended in the fluid conveyed from the second purification chamber (470b) through the second or third purification bead filters (770b, 770c) can be filtered out of such fluid and remain in the second purification chamber (470b); such purification beads can then be conveyed from the second purification chamber (470b) to the first purification chamber (470a) via purification bridging channels (760a, 760b), and then removed from the first purification chamber (470a) via purification bead outlet channel (712d). In this way, purification beads can be removed from any purified fluid leaving the purification chambers (470a, 470b), and thus purification beads can be prevented from interfering with the fluid flow and / or processing downstream of the purification bead filters (770a, 770b, 770c).

[0119] Although this example only shows the delivery of purification beads to / from purification chambers (470a, 470b) for the treatment of fluids therein, it should be understood that, as needed, purification beads or other types of functionalized particles may also be delivered to / from any one or more of other processing chambers (450, 460) for the treatment of fluids therein. For example, any one or more of the processing chambers (450, 460) may be equipped with corresponding bead channels similar to the purification bead channels (712a, 712b, 712c, 712d, 712e), configured to provide a path for conveying the corresponding beads (e.g., together with a liquid solution in which the beads may be suspended) to / from the “wet” area of ​​the corresponding processing chamber (450, 460); and having corresponding valves and / or pumps as described herein. In some cases, any one or more of the processing chambers (450, 460) may also be equipped with a corresponding bead filter similar to a purification bead filter (770a, 770b, 770c), which is configured to provide a path for conveying fluid from the corresponding processing chamber (450, 460) to one or more corresponding fluid channels (712) while preventing beads from the corresponding processing chamber (450, 460) to one or more corresponding fluid channels (712), thereby retaining the beads within the corresponding processing chamber (450, 460).

[0120] Although the purification bead filters (770a, 770b, 770c) of this example are each inserted between one and one or more corresponding fluid outlet channels (712) in the processing chamber portion (752) associated with the purification chamber (470a, 470b), such that the purification bead filters (770a, 770b, 770d) are each located at or near the corresponding fluid outlet of the corresponding purification chamber (470a, 470b), it should be understood that one or more purification bead filters (770a, 770b, 770c) may additionally or alternatively be located at or near the corresponding fluid inlet of the corresponding purification chamber (470a, 470b) or other processing chamber (450, 460). For example, one or more purification bead filters (770a, 770b, 770c) may be inserted between corresponding one and one or more corresponding fluid inlet channels (712) in a processing chamber section (752) associated with a processing chamber (450, 460, 470a, 470b). Such purification bead filters (770a, 770b, 770c) may be configured to provide a path for conveying fluid from one or more corresponding fluid channels (712) to the corresponding processing chamber (450, 460, 470a, 470b) while preventing beads from the corresponding processing chamber (450, 460, 470a, 470b) to one or more corresponding fluid channels (712), thereby retaining the beads within the corresponding processing chamber (450, 460, 470a, 470b). The presence of such purification bead filters (770a, 770b, 770c) at or near the fluid inlet can help mitigate any risk of the beads blocking the fluid from reaching the corresponding one of the processing chamber sections (752) associated with the corresponding processing chambers (450, 460, 470a, 470b).

[0121] See you again Figure 8The fourth layer (800) may include an elastomeric film, such as silicone and / or any other suitable one or more materials. As shown, the fourth layer (800) of this example includes a central opening (802) that is sized and configured to receive a protruding area (750) of the third layer (700). In this example, the fourth layer (800) provides a gasket that forms a seal between adjacent areas of the third layer (700) and the fifth layer (900). In this respect, the fifth layer (900) in this example is in the form of a rigid plate. As shown, the fifth layer (900) of this example includes a central opening (902) that is sized and configured to receive a protruding area (750) of the third layer (700). In the example shown, the sixth layer (1000) is sized and configured to lie directly beneath the protruding area (750) of the third layer (700), and the seventh layer (1100) is sized and configured to lie directly beneath the sixth layer (1000). In this respect, in this example, the sixth layer (1000) is in the form of a gap thermal pad, and in this example, the seventh layer (1100) is in the form of a graphite thermal pad.

[0122] It should be understood that any suitable structure or technique can be used to hold the layers (500, 600, 700, 800, 900, 1000, 1100) together.

[0123] Although the purification bead filters (770a, 770b, 770c) of this example are each formed on the upper surface (702) of the third layer (700), it may be desirable to incorporate one or more purification bead filters (770a, 770b, 770c) at least partially into one or more other suitable layers of the processing chip (400), such as the second layer (600). For example, doing so can reduce the cost and / or complexity of manufacturing the third layer (700), such as when the predetermined particle size of the purification beads is less than about 50 µm, in which case it would otherwise be necessary to process very small (e.g., about 25 µm deep or about 10 µm deep) filter channels (772) in the upper surface (702) of the third layer (700).

[0124] Figure 20A portion of another example of a processing chip (1400) that can provide at least some (if not all) of the features and functions described above is shown. Except as further described below, the processing chip (1400) is similar to the processing chip (400) described above. In this respect, the processing chip (1400) of this example includes a first layer (1500), a second layer (1600), and a third layer (1700), which may be similar to the first layer (500), second layer (600), and third layer (700) described above, respectively, except as further described below. Although not shown, the processing chip (1400) may also include any one or more of the following: a fourth layer similar to a fourth layer (800); a fifth layer similar to a fifth layer (900); a sixth layer similar to a sixth layer (1000); a seventh layer similar to a seventh layer (1100); and / or any other suitable features shown and / or described herein in conjunction with the processing chip (400).

[0125] As shown, the first layer (1500) includes a lower surface (1504) similar to the lower surface (504). Although not shown, the first layer (1500) may also include any one or more of the following: an upper surface similar to the upper surface (502); an opening similar to an opening (510, 520); a pneumatic passage similar to a pneumatic passage (522); a valve chamber portion similar to a valve chamber portion (530); a pump chamber portion similar to a pump chamber portion (540); a protruding area similar to a protruding area (550); a processing chamber portion similar to a processing chamber portion (552); and / or any other suitable features shown and / or described herein in conjunction with the first layer (500).

[0126] The second layer (1600) includes a lower flexible membrane (1600a) and an upper flexible membrane (1600b) that are respectively similar to the flexible membranes (600a, 600b). Although not shown, the second layer (1600) may include openings similar to openings (602a, 602b), holes similar to holes (604), and / or any one or more of any other suitable features shown and / or described herein in conjunction with the second layer (600).

[0127] The third layer (1700) includes: an upper surface (1702) similar to an upper surface (702); a lower surface (1704) similar to a lower surface (704); a plurality of fluid channels (1712) similar to fluid channels (712) (one shown), which are formed as recesses in the upper surface (1702); and a plurality of processing chamber portions (1752) similar to processing chamber portions (752) (one shown), which are formed as recesses in the upper surface (1702). Each processing chamber portion (1752) has a sidewall (1753) extending downward from the upper surface (1702) toward a corresponding base plate (1755). In the example shown, the filter inlet (1774) is formed as a recess in the corresponding sidewall (1753). Although not shown, the third layer (1700) may include any one or more of the following: an opening similar to an opening (710); a fluid passage similar to a fluid passage (714); a valve chamber portion similar to a valve chamber portion (730); a pump chamber portion similar to a pump chamber portion (740); a protruding area similar to a protruding area (750); a sidewall passage similar to a sidewall passage (754); a base plate passage similar to a base plate passage (756); a bridging passage similar to a bridging passage (760); and / or any other suitable feature shown and / or described herein in conjunction with the third layer (700).

[0128] The second layer (1600) of this example also includes a plurality of purification bead filters (1670) (one shown). The purification bead filters (1670) may also be referred to as a “sieve plate structure.” Each purification bead filter (1670) is inserted between one and more corresponding fluid channels (1712) (e.g., one or more corresponding fluid inlet or outlet channels) in the processing chamber section (1752). Each purification bead filter (1670) is configured to provide a path for conveying fluid from the corresponding processing chamber section (1752) to one or more corresponding fluid channels (1712) while inhibiting beads from the corresponding processing chamber section (1752) to one or more corresponding fluid channels (1712), thereby retaining the beads within the processing chamber section (1752).

[0129] In this respect, each of the purification bead filters (1670) includes at least one filter channel (1672) formed as a hole (e.g., a slit) through the upper flexible membrane (1600b), a filter inlet (1674) formed as a first groove through the lower flexible membrane (1600a) and fluidly connecting the inner end of the at least one filter channel (1672) to a filter inlet (1774), and a filter outlet (1676) formed as a second groove through the lower flexible membrane (1600a) and fluidly connecting the outer end of the at least one filter channel (1672) to one or more corresponding fluid channels (1712), such that each filter channel (1672) extends along the upper surface of the lower flexible membrane (1600a) between the corresponding filter inlet (1674) and filter outlet (1676). Each filter channel (1672) is further configured to define the space between the second layer (1600) (e.g., the flexible membrane (1600a) below it) and the first layer (1500), such that fluid can be delivered along the filter channel (1672) and the filter outlet (1676) to one or more corresponding fluid channels (1712).

[0130] Each filter channel (1672) is sized and configured to allow fluid to flow through it while preventing beads with a predetermined particle size (e.g., diameter) from flowing through it. In other words, the filter channels (1672) of each purification bead filter (1670) can be configured to cooperate with each other to act as a “sieve” to filter out these beads from the fluid flowing through it. For this purpose, each filter channel (1672) may have a cross-sectional dimension (e.g., width, depth, diameter, etc.) that is substantially smaller than the corresponding cross-sectional dimensions of some or all of the fluid channels (1712) formed in the upper surface (1702). For example, each filter channel (1672) may be relatively shallow, at least compared to some or all of the fluid channels (1712) formed in the upper surface (1702). In some cases, each filter channel (1672) may have at least one cross-sectional dimension that is substantially smaller than the predetermined particle size of each bead. In the case where each bead has a predetermined particle size of about 50 µm, at least one cross-sectional dimension of each filter channel (1672) may be substantially smaller than about 50 µm. For example, each filter channel (1672) may have a depth in the range of about 10 µm to about 30 µm (e.g., relative to the lower surface (1504)), such as a depth of about 10 µm or a depth of about 25 µm. As shown, the depth of each filter channel (1672) may be substantially equal to the thickness of the upper flexible membrane (1600b). Thus, with the upper flexible membrane (1600b) having a thickness of about 10 µm, each filter channel (1672) may have a depth of about 10 µm. Since one cross-sectional dimension of each filter channel (1672) (e.g., its depth) may be small enough to prevent beads from flowing through it, another cross-sectional dimension of each filter channel (1672) (e.g., its width) may in some cases be substantially equal to or even greater than the predetermined particle size of each bead. Alternatively, each filter channel (1672) may have any other suitable cross-sectional dimensions, such as any other suitable depth, selected to filter out beads of any predetermined particle size.

[0131] The presence of a filter inlet (1774) in each purification bead filter (1670) helps mitigate any risk of beads clogging the fluid flow to the corresponding filter channel (1672) by providing space for the beads to aggregate without obstructing the corresponding filter channel (1672). Alternatively, the filter channels (1672) of each purification bead filter (1670) may have a sufficiently large width to reduce the impact of any blockages that may occur in the filter channels (1672) on fluid flow by providing one or more alternative paths for the fluid to reach the corresponding filter outlet (1676) (e.g., by bypassing blockages within the filter channels (1672)). It should be understood that each purification bead filter (1670) may include any suitable number of filter channels (1672).

[0132] Particles can move along a path similar to Figure 19 The flow path shown flows through the processing chip (1400). It is understood that the second layer (1600) can be pneumatically deformed in a similar manner to that described above to provide appropriate valved and / or peristaltic pumping for delivering the purification beads along each section of the flow path.

[0133] Because a purification bead filter (1670) is present at the fluid outlet of the illustrated processing chamber section (1752), any purification beads that may be suspended in the fluid conveyed from the illustrated processing chamber section (1752) through the illustrated purification bead filter (1670) can be filtered out of this fluid and remain in the illustrated processing chamber section (1752). In this way, purification beads can be removed from any purified fluid leaving the illustrated processing chamber section (1752), thus preventing purification beads from interfering with the fluid flow and / or processing downstream of the illustrated purification bead filter (1670).

[0134] Figure 21A portion of another example of a processing chip (2400) that can provide at least some (if not all) of the features and functions described above is shown. Except as further described below, the processing chip (2400) is similar to the processing chip (400) described above. In this respect, the processing chip (2400) of this example includes a first layer (not shown) similar to a first layer (500), a second layer (not shown) similar to a second layer (600), and a third layer (2700), which, except as further described below, may be similar to the third layer (700) described above. Although not shown, the processing chip (2400) may also include any one or more of the following: a fourth layer similar to a fourth layer (800); a fifth layer similar to a fifth layer (900); a sixth layer similar to a sixth layer (1000); a seventh layer similar to a seventh layer (1100); and / or any other suitable features shown and / or described herein in conjunction with the processing chip (400).

[0135] As shown in the figure, the third layer (2700) includes: an upper surface (2702) similar to an upper surface (702); a plurality of fluid channels (2712) similar to fluid channels (712), which are formed as recesses in the upper surface (2702); a plurality of valve chamber portions (2730) similar to valve chamber portions (730), which are formed as recesses in the upper surface (2702); and a plurality of processing chamber portions (2752) similar to processing chamber portions (752), which are formed as recesses in the upper surface (2702). Each processing chamber portion (2752) has a sidewall (2753) extending downward from the upper surface (2702) toward a corresponding base plate (2755). Furthermore... Figure 21 As seen, the third layer (2700) includes a sidewall channel (2754) similar to a sidewall channel (754) and a base plate channel (2756) similar to a base plate channel (756). Although not shown, the third layer (2700) may include: an opening similar to an opening (710); a fluid channel similar to a fluid channel (714); a pump chamber portion similar to a pump chamber portion (740); a protruding area similar to a protruding area (750); a bridging channel similar to a bridging channel (760); and / or any one or more of any other suitable features shown and / or described herein in conjunction with the third layer (700).

[0136] In the example shown, the plurality of fluid passages (2712) include at least one fluid inlet passage (2712a) leading to a respective processing chamber section (2752) for conveying fluid to the respective processing chamber section (2752), at least one fluid outlet passage (2712b) for conveying fluid from the respective processing chamber section (2752), and at least one backflow passage (2712c), the purpose of which will be described below. Also in this example, the plurality of valve chamber sections (2730) include at least one fluid inlet valve chamber section (2730a) upstream of at least one fluid inlet passage (2712a), at least one fluid outlet valve chamber section (2730b) downstream of at least one fluid outlet passage (2712b), and at least one backflow valve chamber section (2730c) positioned along at least one backflow passage (2712c).

[0137] The third layer (2700) of this example also includes a plurality of purification bead filters (2770a, 2770b) on the upper surface (2702). The purification bead filters (2770a, 2770b) may also be referred to as a “sieve plate structure.” Each purification bead filter (2770a, 2770b) is inserted between one or more corresponding fluid outlet channels (2712b) in the processing chamber section (2752). Each purification bead filter (2770a, 2770b) is configured to provide a path for conveying fluid from the corresponding processing chamber section (2752) to one or more corresponding fluid outlet channels (2712b) while preventing beads from flowing from the corresponding processing chamber section (2752) to one or more corresponding fluid outlet channels (2712b), thereby retaining the beads within the processing chamber section (2752).

[0138] In this respect, each of the purification bead filters (2770a, 2770b) includes an array of filter channels (2772) formed as recesses in the upper surface (2702), a filter inlet (2774) formed as recesses in the respective sidewalls (2753) and extending between the inner ends of the respective filter channels (2772), and a filter outlet (2776) formed as recesses in the upper surface (2702) and extending between the outer ends of the respective filter channels (2772), such that each filter channel (2772) extends between the respective filter inlet (2774) and filter outlet (2776). Each filter channel (2772) is further configured to define the space between the third layer (2700) and the second layer (600), and each filter outlet (2776) is similarly configured to define the space between the third layer (2700) and the second layer (600), such that fluid can be conveyed along the filter channel (2772) and the filter outlet (2776) to one or more corresponding fluid outlet channels (2712b).

[0139] Each filter channel (2772) can be individually sized and configured to allow fluid flow through it while inhibiting the flow of beads with a predetermined particle size (e.g., diameter). In other words, the filter channels (2772) of each purification bead filter (2770a, 2770b) can be configured to cooperate with each other and / or with the beads themselves to act as a sieve or "sieve plate" to filter out the beads from the fluid flowing through it. For this purpose, each filter channel (2772) can have a cross-sectional dimension (e.g., width, depth, diameter, etc.) that is substantially smaller than the corresponding cross-sectional dimension of some or all of the fluid channels (2712) formed in the upper surface (2702). While each individual filter channel (2772) can have a cross-sectional area smaller than some or all of the cross-sectional area of ​​the fluid channels (2712), the multiple filter channels (2772) of a particular purification bead filter (2770a, 2770b) can have a total cross-sectional area that is substantially equal to or greater than some or all of the cross-sectional area of ​​the fluid channels (2712). For example, the filter channels (2772) may each be relatively shallow compared to at least some or all of the fluid channels (2712) formed in the upper surface (2702). In some cases, the filter channels (2772) may each have at least one cross-sectional dimension substantially larger than the predetermined particle size of each bead. For example, the filter channels (2772) may each have at least one cross-sectional dimension that is greater than or equal to about twice the predetermined particle size of each bead and / or less than about five times the predetermined particle size of each bead. In the case where each bead has a predetermined particle size of about 50 µm, at least one cross-sectional dimension of each filter channel (2772) may be substantially larger than about 50 µm. For example, each filter channel (2772) may have a depth greater than or equal to about 100 µm (e.g., relative to the upper surface (2702)). Even if one cross-sectional dimension (e.g., its depth) of each filter channel (2772) is substantially larger than the predetermined particle size of each bead, another cross-sectional dimension (e.g., its width) of each filter channel (2772) may also be substantially larger than the predetermined particle size of each bead. With each bead having a predetermined particle size of about 50 µm, another cross-sectional dimension of each filter channel (2772) can be substantially equal to or greater than about 50 µm. For example, each filter channel (2772) can have a width in the range of about 100 µm to about 400 µm, such as a width of about 200 µm.

[0140] Therefore, the filter channel (2772) can be substantially larger than the beads. However, the filter channel (2772) can be configured to prevent the beads from flowing through it by blocking at least some of the beads therein. This blocking can help suppress the flow of beads while still allowing fluid to flow through the filter channel (2772), for example, where there is some variation in size between the beads, i.e., where each bead has a predetermined particle size with an acceptable degree of variation. For example, each bead may have a predetermined particle size of 50µm ± 10µm. In this case, larger beads can cooperate with smaller beads to block within the filter channel (2772), while defining openings between the beads that are large enough to allow fluid to flow through and small enough to prevent other beads from flowing through. Thus, although each individual bead is substantially smaller than the filter channel (2772), the beads can be retained within the processing chamber portion (2752). In some cases, substantially a very small number of beads are able to escape from the filter channel (2772). For example, substantially a minimum amount of beads can escape from the filter channel (2772) before a blockage forms in it. Furthermore, or alternatively, after blockage forms in the filter channel (2772), essentially a minimal amount of beads may be able to escape from the filter channel (2772) through one or more openings defined by the blockage. However, due to the bead blockage within the filter channel (2772), only a single-digit percentage (e.g., only about 1%) of the beads contained within the processing chamber section (2752) is able to escape from the processing chamber section (2752) through the purification bead filters (2770a, 2770b). It should also be understood that the increased size of the filter channel (2772) relative to the beads can help mitigate any risk of bead blockage of fluid flow through each filter channel (2772).

[0141] In the example shown, the fluid path downstream of the filter channel (2772) is substantially free of potential clogging areas to avoid clogging by any beads that might escape from the respective filter channel (2772). For example, the processing chip (2400) in this example has no vacuum cap between each purification bead filter (2770a, 2770b) and the corresponding fluid outlet valve chamber section (2730b). Alternatively, the dimensions of each filter outlet (2776) and / or each fluid outlet channel (2712b) can be designed to avoid clogging by any beads that might escape from the respective filter channel (2772). For example, each cross-sectional dimension (e.g., width and depth) of each filter outlet (2776) and / or each fluid outlet channel (2712b) can be approximately 5 times larger than the predetermined particle size of each bead. With each bead having a predetermined particle size of approximately 50 µm, each filter outlet (2776) and / or each fluid outlet channel (2712b) may have a width of approximately 500 µm and / or a depth of approximately 250 µm.

[0142] Similarly, each fluid inlet channel (2712a) can be sized to avoid clogging any beads that might flow upstream from the corresponding processing chamber section (2752). For example, each cross-sectional dimension (e.g., width and depth) of each fluid inlet channel (2712a) can be approximately 5 times larger than the predetermined particle size of each bead. With each bead having a predetermined particle size of approximately 50 µm, each fluid inlet channel (2712a) can have a width of approximately 500 µm and / or a depth of approximately 250 µm. Thus, the fluid inlet channel (2712a) can have increased dimensions relative to at least some of the other fluid channels (2712) on the third layer (2700) (such as the fluid channel (2712) upstream of the corresponding fluid inlet valve chamber section (2730a)).

[0143] In the illustrated example, each backflush channel (2712c) is downstream of the corresponding filter outlet (2776), such that each backflush channel (2712c) is configured to facilitate backflushing of the corresponding filter channel (2772), as in the case that one or more of the corresponding filter channels (2772) are substantially clogged with beads. For example, with the valve associated with the backflush valve chamber section (2730c) in the open position and the valve associated with the fluid outlet valve chamber section (2730b) in the closed position, backflush fluid (e.g., water) can flow upstream from the backflush channel (2712c) into the corresponding filter outlet (2776) and through the corresponding filter channel (2772) to push any beads that may be clogged within the corresponding filter channel (2772) back into the corresponding processing chamber section (2752). This backflushing of the filter channels (2772) can be performed according to predetermined routines based on certain criteria. For example, such backwashing can be performed in response to the duration of fluid flowing through the respective filters (2770a, 2770b) reaching a predetermined threshold duration; and / or in response to the cumulative volume of fluid flowing through the respective filters (2770a, 2770b) reaching a predetermined threshold volume; and / or in response to the number of cycles of fluid flowing through the respective filters (2770a, 2770b) reaching a predetermined threshold number of cycles (e.g., one cycle, two cycles, or any other suitable number of cycles); and / or in response to pressure data indicating flow blockage within the respective filters (2770a, 2770b).

[0144] Alternatively, each backflush channel (2712c) may be configured to facilitate flushing of the entire fluid path from the corresponding processing chamber section (2752) to the corresponding target vial or other fluid storage container (e.g., in the reagent storage frame (107)). For example, when the valve associated with the backflush valve chamber section (2730c) is open, the valve associated with the fluid outlet valve chamber section (2730b) is open, and the processing chamber associated with the processing chamber section (2752) is closed, backflush fluid (e.g., water) can flow downstream from the backflush channel (2712c) into the corresponding fluid outlet channel (2712b) and through the corresponding fluid outlet valve chamber section (2730b) all the way to the corresponding target vial / container to push any beads or fluid set along the fluid path toward the corresponding target vial / container.

[0145] VI. Combined Examples The following examples illustrate various non-exhaustive ways in which the teachings of this document can be combined or applied. It should be understood that the following examples are not intended to limit the scope of any claim that may appear at any time in this application or in subsequent documents thereof. No waiver of rights is intended. The following examples are provided merely for illustrative purposes. It is contemplated that the various teachings of this document may be arranged and applied in a variety of other ways. It is also contemplated that some variations may omit certain features mentioned in the examples below. Therefore, no aspect or feature mentioned below should be considered critical unless explicitly stated later by the inventor or a successor with an interest in the inventor. If any claim set forth in this application or in subsequent documents related to this application includes additional features beyond those mentioned below, such additional features shall not be presumed to have been added for any reason relating to patentability.

[0146] Example 1 A fluid device includes: (a) a first layer defining a first chamber portion configured to receive pressurized gas; (b) a second layer defining: (i) a second chamber portion located below the first chamber portion, the second chamber portion being configured to receive at least one liquid and a plurality of particles; (ii) at least one fluid outlet channel in fluid communication with the second chamber portion; and (iii) a filter formed in a surface of the second layer between the second chamber portion and the at least one fluid outlet channel; and (c) an elastic layer disposed between the first layer and the second layer, the elastic layer being deformable into the second chamber portion to drive the at least one liquid out of the second chamber portion through the filter and into the at least one fluid outlet channel, the filter being configured to allow the at least one liquid to flow through it and to prevent the plurality of particles from flowing through it.

[0147] Example 2 According to the fluid apparatus described in Example 1, the filter includes a plurality of filter channels.

[0148] Example 3 According to the fluid apparatus described in Example 2, each of the plurality of filter channels is shallower than the at least one fluid outlet channel.

[0149] Example 4 According to any one of Examples 2 to 3, each of the plurality of filter channels has a depth of substantially less than about 50 µm.

[0150] Example 5 According to the fluid apparatus described in Example 4, each of the plurality of filter channels has a depth of approximately 25 µm.

[0151] Example 6 According to the fluid apparatus described in Example 4, the depth of each of the plurality of filter channels ranges from about 10 µm to about 23 µm.

[0152] Example 7 According to any one of Examples 2 to 6, each of the plurality of filter channels extends between a corresponding inner end located near the second chamber portion and a corresponding outer end located away from the second chamber portion.

[0153] Example 8 According to the fluid apparatus of Example 7, the filter further includes a filter inlet extending between the inner ends of the plurality of filter channels.

[0154] Example 9 According to any one of Examples 7 to 8, the fluid apparatus further includes a filter outlet extending between the outer ends of the plurality of filter channels.

[0155] Example 10 According to any one of Examples 1 to 9, the fluid device has a second chamber portion having a sidewall, and the second layer defines at least one sidewall passage extending along the sidewall.

[0156] Example 11 According to the fluid device of Example 10, the at least one sidewall channel extends along the sidewall to the filter.

[0157] Example 12 According to the fluid device of Example 11, the at least one sidewall channel includes a plurality of sidewall channels extending along the sidewall to the filter.

[0158] Example 13 According to any one of Examples 10 to 12, the fluid apparatus has a second chamber portion having a base plate, and the second layer defines a base plate channel extending along the base plate.

[0159] Example 14 According to the fluid device of Example 13, the at least one sidewall channel extends along the sidewall to the bottom plate channel.

[0160] Example 15 According to any one of Examples 10 to 14, the fluid device has at least one sidewall channel with a width of about 500 µm.

[0161] Example 16 According to any one of Examples 10 to 15, the fluid device has at least one sidewall channel with a depth of about 100 µm.

[0162] Example 17 According to any one of Examples 1 to 16, the second layer further defines a particle inlet channel in fluid communication with the second chamber portion, the particle inlet channel being configured to deliver the plurality of particles to the second chamber portion.

[0163] Example 18 According to the fluid apparatus of Example 17, at least one cross-sectional dimension of the particle inlet channel is larger than the corresponding cross-sectional dimension of the at least one fluid outlet channel.

[0164] Example 19 According to any one of Examples 17 to 18, the fluid device has a particle inlet channel with a width of about 500 µm.

[0165] Example 20 According to any one of Examples 17 to 19, the fluid device has a particle inlet channel with a depth of about 250 µm.

[0166] Example 21 According to any one of Examples 1 to 20, the first layer further defines a third chamber portion configured to receive pressurized gas, and the second layer further defines: (i) a fourth chamber portion located below the third chamber portion, the fourth chamber portion being configured to receive the at least one liquid and the plurality of particles, and (ii) at least one bridging channel extending between the second chamber portion and the fourth chamber portion, the at least one bridging channel being configured to provide a path for conveying the at least one liquid and the plurality of particles between the second chamber portion and the fourth chamber portion.

[0167] Example 22 According to the fluid device described in Example 21, the at least one bridging channel has a width of about 500 µm.

[0168] Example 23 According to any one of Examples 21 to 22, the fluid device has at least one bridging channel with a depth of about 500 µm.

[0169] Example 24 According to any one of Examples 1 to 23, the second layer further defines a particle outlet channel in fluid communication with the second chamber portion, the particle outlet channel being configured to remove the plurality of particles from the second chamber portion.

[0170] Example 25 According to the fluid apparatus described in Example 24, at least one cross-sectional dimension of the particle outlet channel is larger than the corresponding cross-sectional dimension of the at least one fluid outlet channel.

[0171] Example 26 A fluid device includes: (a) a first layer defining a first chamber portion configured to receive pressurized gas; (b) a second layer defining: (i) a second chamber portion located below the first chamber portion, the second chamber portion being configured to receive at least one liquid and a plurality of particles; (ii) a fluid outlet passage in fluid communication with the second chamber portion; and (iii) a plurality of filter passages located between the second chamber portion and the fluid outlet passage; and (c) an elastic layer disposed between the first layer and the second layer, the elastic layer being deformable into the second chamber portion to drive the at least one liquid out of the second chamber portion through the plurality of filter passages and into the fluid outlet passage, the plurality of filter passages being configured to allow the at least one liquid to flow through therethrough and the plurality of filter passages being configured to prevent the plurality of particles from flowing through therethrough.

[0172] Example 27 According to the fluid apparatus of Example 26, each of the plurality of filter channels extends between a corresponding inner end located near the second chamber portion and a corresponding outer end located away from the second chamber portion.

[0173] Example 28 According to the fluid apparatus described in Example 27, the second layer further defines a filter inlet extending between the inner ends of the plurality of filter channels.

[0174] Example 29 According to any one of Examples 26 to 27, the fluid apparatus, the second layer further defines a filter outlet extending between the outer ends of the plurality of filter channels.

[0175] Example 30 A fluid device includes: (a) a first layer defining a first chamber portion configured to receive pressurized gas; (b) a second layer defining: (i) a second chamber portion located below the first chamber portion, the second chamber portion having sidewalls and configured to receive at least one liquid and a plurality of particles; (ii) a fluid outlet passage in fluid communication with the second chamber portion; (iii) a filter located between the second chamber portion and the at least one fluid outlet passage, the filter being formed in a surface of the second layer; and (iv) a plurality of sidewall passages extending along the sidewalls to the filter; and (c) an elastic layer disposed between the first layer and the second layer, the elastic layer being deformable into the second chamber portion to expel the at least one liquid from the second chamber portion via the plurality of sidewall passages, through the filter, and into the fluid outlet passage, the filter being configured to allow the at least one liquid to flow through it and to prevent the plurality of particles from flowing through it.

[0176] Example 31 A method of using an apparatus, the apparatus comprising: (a) a first layer defining a first chamber portion configured to receive pressurized gas; (b) a second layer defining: (i) a second chamber portion located below the first chamber portion, the second chamber portion being configured to receive at least one liquid and a plurality of particles; (ii) at least one fluid outlet channel in fluid communication with the second chamber portion; and (iii) a filter between the second chamber portion and the at least one fluid outlet channel; and (c) an elastic layer disposed between the first layer and the second layer; the method comprising treating the at least one liquid with the plurality of particles within the second chamber portion.

[0177] Example 32 According to the method of Example 31, the at least one liquid contains an RNA therapeutic agent.

[0178] Example 33 According to any one of Examples 31 to 32, the plurality of particles comprises a plurality of beads.

[0179] Example 34 According to any one of Examples 31 to 33, the plurality of particles includes a plurality of purified particles, and the step of processing the at least one liquid includes purifying the at least one liquid using the plurality of purified particles.

[0180] Example 35 According to the method described in Example 34, each of the plurality of purified particles is configured to separate mRNA from one or more components of the transcription reaction process.

[0181] Example 36 According to the method of Example 35, the one or more components include at least one of plasmid DNA or an enzyme.

[0182] Example 37 According to any one of Examples 35 to 36, each of the plurality of purified particles is configured to separate mRNA from the components of the transcription reaction process by selectively capturing mRNA via a polyadenylate (polyA) tail using a salt and water purification step.

[0183] Example 38 According to any one of Examples 34 to 37, each of the plurality of purified particles includes a rigid polymer resin support matrix.

[0184] Example 39 According to the method of Example 38, the rigid polymer resin support matrix comprises crosslinked poly(styrene-divinylbenzene).

[0185] Example 40 According to any one of Examples 34 to 39, each of the plurality of purified particles has a multi-hydroxyl surface coating.

[0186] Example 41 According to any one of Examples 34 to 40, each of the plurality of purified particles has a surface functionalized with poly(dT).

[0187] Example 42 According to any one of Examples 31 to 41, each of the plurality of particles has a predetermined particle size.

[0188] Example 43 According to the method described in Example 42, the predetermined particle size is approximately 50 µm.

[0189] Example 44 According to any one of Examples 31 to 43, each of the plurality of particles is porous.

[0190] Example 45 The method according to any one of Examples 31 to 44 further includes deforming the elastic layer into the second chamber portion, thereby driving the at least one liquid out of the second chamber portion through the filter and into the at least one fluid outlet channel.

[0191] Example 46 According to the method of Example 45, the filter allows the at least one liquid to flow through it during the deformation step, and the filter prevents the plurality of particles from flowing through it during the deformation step.

[0192] Example 47 According to any one of Examples 31 to 46, the second layer further defines a particle inlet channel in fluid communication with the second chamber portion, and the method further includes delivering the plurality of particles to the second chamber portion via the particle inlet channel.

[0193] Example 48 According to any one of Examples 31 to 47, the first layer further defines a third chamber portion configured to receive pressurized gas, and the second layer further defines: (i) a fourth chamber portion located below the third chamber portion, the fourth chamber portion being configured to receive the at least one liquid and the plurality of particles, and (ii) at least one bridging channel extending between the second chamber portion and the fourth chamber portion; the method further includes conveying the at least one liquid and the plurality of particles between the second chamber portion and the fourth chamber portion via the at least one bridging channel.

[0194] Example 49 According to any one of Examples 31 to 48, the second layer further defines a particle outlet channel in fluid communication with the second chamber portion, and the method further includes removing the plurality of particles from the second chamber portion via the particle outlet channel.

[0195] Example 50 A method of using an apparatus, the apparatus comprising: (a) a first layer defining a first chamber portion configured to receive pressurized gas; (b) a second layer defining: (i) a second chamber portion located below the first chamber portion, the second chamber portion being configured to receive at least one liquid and a plurality of particles; (ii) a fluid outlet passage in fluid communication with the second chamber portion; and (iii) a plurality of filter passages located between the second chamber portion and the fluid outlet passage; and (c) an elastic layer disposed between the first layer and the second layer; the method comprising treating the at least one liquid with the plurality of particles within the second chamber portion.

[0196] Example 51 According to the method of Example 50, the at least one liquid contains an RNA therapeutic agent.

[0197] Example 52 According to any one of Examples 50 to 51, the plurality of particles comprises a plurality of beads.

[0198] Example 53 According to any one of Examples 50 to 52, the plurality of particles comprises a plurality of purified particles, and the step of processing the at least one liquid includes purifying the at least one liquid using the plurality of purified particles.

[0199] Example 54 According to the method described in Example 53, each of the plurality of purified particles is configured to separate mRNA from one or more components of the transcription reaction process.

[0200] Example 55 According to the method of Example 54, the one or more components include at least one of plasmid DNA or an enzyme.

[0201] Example 56 According to any one of Examples 54 to 55, each of the plurality of purified particles is configured to separate mRNA from the components of the transcription reaction process by selectively capturing mRNA via a polyadenylate (polyA) tail using a salt and water purification step.

[0202] Example 57 According to any one of Examples 53 to 56, each of the plurality of purified particles includes a rigid polymer resin support matrix.

[0203] Example 58 According to the method of Example 57, the rigid polymer resin support matrix comprises crosslinked poly(styrene-divinylbenzene).

[0204] Example 59 According to any one of Examples 53 to 58, each of the plurality of purified particles has a multi-hydroxyl surface coating.

[0205] Example 60 According to any one of Examples 53 to 59, each of the plurality of purified particles has a surface functionalized with poly(dT).

[0206] Example 61 According to the method of any one of Examples 50 to 60, each of the plurality of particles has a predetermined particle size.

[0207] Example 62 According to the method described in Example 61, the predetermined particle size is about 50 µm.

[0208] Example 63 According to any one of Examples 50 to 62, each of the plurality of particles is porous.

[0209] Example 64 The method according to any one of Examples 50 to 63 further includes deforming the elastic layer into the second chamber portion, thereby displacing the at least one liquid out of the second chamber portion through the plurality of filter channels and into the fluid outlet channel.

[0210] Example 65 According to the method of Example 64, the plurality of filter channels allow the at least one liquid to flow through it during the deformation step, and the plurality of filter channels prevent the plurality of particles from flowing through it during the deformation step.

[0211] Example 66 According to any one of Examples 50 to 65, the second layer further defines a particle inlet channel in fluid communication with the second chamber portion, and the method further includes delivering the plurality of particles to the second chamber portion via the particle inlet channel.

[0212] Example 67 According to any one of Examples 50 to 66, the first layer further defines a third chamber portion configured to receive pressurized gas, and the second layer further defines: (i) a fourth chamber portion located below the third chamber portion, the fourth chamber portion being configured to receive the at least one liquid and the plurality of particles, and (ii) at least one bridging channel extending between the second chamber portion and the fourth chamber portion; the method further includes conveying the at least one liquid and the plurality of particles between the second chamber portion and the fourth chamber portion via the at least one bridging channel.

[0213] Example 68 According to any one of Examples 50 to 67, the second layer further defines a particle outlet channel in fluid communication with the second chamber portion, and the method further includes removing the plurality of particles from the second chamber portion via the particle outlet channel.

[0214] Example 69 A method of using an apparatus, the apparatus comprising: (a) a first layer defining a first chamber portion configured to receive pressurized gas; (b) a second layer defining: (i) a second chamber portion located below the first chamber portion, the second chamber portion having sidewalls and configured to receive at least one liquid and a plurality of particles; (ii) a fluid outlet passage in fluid communication with the second chamber portion; (iii) a filter located between the second chamber portion and the at least one fluid outlet passage; and (iv) a plurality of sidewall passages extending along the sidewalls to the filter; and (c) an elastic layer disposed between the first layer and the second layer; the method comprising treating the at least one liquid with the plurality of particles within the second chamber portion.

[0215] Example 70 According to the method described in Example 69, the at least one liquid contains an RNA therapeutic agent.

[0216] Example 71 According to any one of Examples 69 to 70, the plurality of particles comprises a plurality of beads.

[0217] Example 72 According to any one of Examples 69 to 71, the plurality of particles comprises a plurality of purified particles, and the step of processing the at least one liquid includes purifying the at least one liquid using the plurality of purified particles.

[0218] Example 73 According to the method described in Example 72, each of the plurality of purified particles is configured to isolate mRNA from components of the transcription reaction process.

[0219] Example 74 According to the method of Example 73, the component includes at least one of plasmid DNA or an enzyme.

[0220] Example 75 According to any one of Examples 73 to 74, each of the plurality of purified particles is configured to separate mRNA from the components of the transcription reaction process by selectively capturing mRNA via a polyadenylate (polyA) tail using a salt and water purification step.

[0221] Example 76 According to any one of Examples 72 to 75, each of the plurality of purified particles comprises a rigid polymer resin support matrix.

[0222] Example 77 According to the method of Example 76, the rigid polymer resin support matrix comprises crosslinked poly(styrene-divinylbenzene).

[0223] Example 78 According to any one of Examples 72 to 77, each of the plurality of purified particles has a multi-hydroxyl surface coating.

[0224] Example 79 According to any one of Examples 72 to 78, each of the plurality of purified particles has a surface functionalized with poly(dT).

[0225] Example 80 According to any one of Examples 69 to 79, each of the plurality of particles has a predetermined particle size.

[0226] Example 81 According to the method described in Example 80, the predetermined particle size is approximately 50 µm.

[0227] Example 82 According to any one of Examples 69 to 81, each of the plurality of particles is porous.

[0228] Example 83 The method according to any one of Examples 69 to 82 further includes deforming the elastic layer into the second chamber portion to expel the at least one liquid from the second chamber portion, through the filter, and into the fluid outlet channel via the plurality of sidewall channels.

[0229] Example 84 According to the method of Example 83, the filter allows the at least one liquid to flow through it during the deformation step, and the filter prevents the plurality of particles from flowing through it during the deformation step.

[0230] Example 85 According to any one of Examples 69 to 84, the second layer further defines a particle inlet channel in fluid communication with the second chamber portion, and the method further includes delivering the plurality of particles to the second chamber portion via the particle inlet channel.

[0231] Example 86 According to any one of Examples 69 to 85, the first layer further defines a third chamber portion configured to receive pressurized gas, and the second layer further defines: (i) a fourth chamber portion located below the third chamber portion, the fourth chamber portion being configured to receive the at least one liquid and the plurality of particles, and (ii) at least one bridging channel extending between the second chamber portion and the fourth chamber portion; the method further includes conveying the at least one liquid and the plurality of particles between the second chamber portion and the fourth chamber portion via the at least one bridging channel.

[0232] Example 87 According to any one of Examples 69 to 86, the second layer further defines a particle outlet channel in fluid communication with the second chamber portion, and the method further includes removing the plurality of particles from the second chamber portion via the particle outlet channel.

[0233] Example 88 A fluid device includes: (a) a first layer defining a first chamber portion configured to receive pressurized gas; (b) a second layer defining: (i) a second chamber portion located below the first chamber portion, the second chamber portion being configured to receive at least one liquid and a plurality of particles; and (ii) at least one fluid channel in fluid communication with the second chamber portion; (c) an elastic layer disposed between the first layer and the second layer, the elastic layer being deformable into the second chamber portion to expel the at least one liquid from the second chamber portion; and (d) a filter between the second chamber portion and the at least one fluid channel, the filter being formed in a surface of at least one of the second layer or the elastic layer, the filter being configured to allow the at least one liquid to flow through it and to prevent the plurality of particles from flowing through it.

[0234] Example 89 According to the fluid apparatus described in Example 88, the filter includes at least one filter channel.

[0235] Example 90 According to the fluid device described in Example 89, the at least one filter channel is shallower than the at least one fluid channel.

[0236] Example 91 According to any one of Examples 89 to 90, the depth of the at least one filter channel is substantially less than about 50 µm.

[0237] Example 92 According to the fluid apparatus described in Example 91, the at least one filter channel has a depth of about 25 µm.

[0238] Example 93 According to the fluid apparatus described in Example 91, the at least one filter channel has a depth of about 10 µm.

[0239] Example 94 According to any one of Examples 88 to 93, the filter is formed in at least one surface of the second layer.

[0240] Example 95 According to any one of Examples 88 to 93, the filter is formed in at least one surface of the elastic layer in the fluid device.

[0241] Example 96 According to the fluid device described in Example 95, the elastic layer comprises an upper membrane and a lower membrane.

[0242] Example 97 According to the fluid apparatus described in Example 96, the filter includes pores extending through the upper membrane.

[0243] Example 98 According to the fluid device described in Example 97, the orifice defines a filter channel between the first layer and the lower membrane.

[0244] Example 99 According to any one of Examples 96 to 98, the fluid apparatus includes a first groove and a second groove extending through the lower membrane.

[0245] Example 100 According to the fluid apparatus described in Example 99, the first tank and the second tank define a filter inlet and a filter outlet, respectively.

[0246] Example 101 According to the fluid apparatus of Example 100, the filter inlet is in partial fluid communication with the second chamber, and the filter outlet is in fluid communication with the at least one fluid channel.

[0247] Example 102 According to any one of Examples 96 to 101, the fluid device has an upper membrane with a thickness of about 25 µm.

[0248] Example 103 According to any one of Examples 96 to 101, the upper membrane has a thickness of about 10 µm.

[0249] Example 104 According to any one of Examples 88 to 103, the at least one fluid passage includes at least one fluid inlet passage.

[0250] Example 105 According to any one of Examples 88 to 104, the at least one fluid passage includes at least one fluid outlet passage.

[0251] Example 106 A fluid device includes: (a) a first layer defining a first chamber portion configured to receive pressurized gas; (b) a second layer defining: (i) a second chamber portion located below the first chamber portion, the second chamber portion being configured to receive at least one liquid and a plurality of particles; and (ii) a fluid channel in fluid communication with the second chamber portion; (c) an elastic layer disposed between the first layer and the second layer, the elastic layer being deformable into the second chamber portion to expel the at least one liquid from the second chamber portion; and (d) at least one filter channel located between the second chamber portion and the fluid channel, the at least one filter channel being formed in a surface of at least one of the second layer or the elastic layer, the at least one filter channel being configured to allow the at least one liquid to flow through it, and the at least one filter channel being configured to prevent the plurality of particles from flowing through it.

[0252] Example 107 According to the fluid device of Example 106, the at least one filter channel is formed in at least one surface of the elastic layer.

[0253] Example 108 According to the fluid device described in Example 107, the elastic layer comprises an upper membrane and a lower membrane.

[0254] Example 109 According to the fluid apparatus of Example 108, the at least one filter channel is defined by a pore extending through the upper membrane.

[0255] Example 110 A fluid device includes: (a) a first layer defining a first chamber portion configured to receive pressurized gas; (b) a second layer defining: (i) a second chamber portion located below the first chamber portion, the second chamber portion having sidewalls, the second chamber portion being configured to receive at least one liquid and a plurality of particles; (ii) a fluid passage in fluid communication with the second chamber portion; and (iii) a plurality of sidewall passages extending along the sidewalls; (c) an elastic layer disposed between the first layer and the second layer, the elastic layer being deformable into the second chamber portion to expel the at least one liquid from the second chamber portion via the plurality of sidewall passages; and (d) a filter between the second chamber portion and the at least one fluid passage, the filter being formed in a surface of at least one of the second layer or the elastic layer, the filter being configured to allow the at least one liquid to flow through it, the filter being configured to prevent the plurality of particles from flowing through it, the plurality of sidewall passages extending along the sidewalls to the filter.

[0256] Example 111 A method of using an apparatus, the apparatus comprising: (a) a first layer defining a first chamber portion configured to receive pressurized gas; (b) a second layer defining: (i) a second chamber portion located below the first chamber portion, the second chamber portion being configured to receive at least one liquid and a plurality of particles; (ii) at least one fluid channel in fluid communication with the second chamber portion; (c) an elastic layer disposed between the first layer and the second layer; and (d) a filter between the second chamber portion and the at least one fluid channel, the filter being defined by at least one of the second layer or the elastic layer; the method comprising treating the at least one liquid with the plurality of particles within the second chamber portion.

[0257] Example 112 According to the method described in Example 111, the at least one liquid contains an RNA therapeutic agent.

[0258] Example 113 According to any one of Examples 111 to 112, the plurality of particles comprises a plurality of beads.

[0259] Example 114 According to any one of Examples 111 to 113, the plurality of particles comprises a plurality of purified particles, and the step of processing the at least one liquid includes purifying the at least one liquid using the plurality of purified particles.

[0260] Example 115 According to the method described in Example 114, each of the plurality of purified particles is configured to separate mRNA from one or more components of the transcription reaction process.

[0261] Example 116 According to the method of Example 115, the one or more components include at least one of plasmid DNA or an enzyme.

[0262] Example 117 According to any one of Examples 115 to 116, each of the plurality of purified particles is configured to separate mRNA from the components of the transcription reaction process by selectively capturing mRNA via a polyadenylate (polyA) tail using a salt and water purification step.

[0263] Example 118 According to any one of Examples 114 to 117, each of the plurality of purified particles comprises a rigid polymer resin support matrix.

[0264] Example 119 According to the method of Example 118, the rigid polymer resin support matrix comprises crosslinked poly(styrene-divinylbenzene).

[0265] Example 120 According to any one of Examples 114 to 119, each of the plurality of purified particles has a multi-hydroxyl surface coating.

[0266] Example 121 According to any one of Examples 114 to 120, each of the plurality of purified particles has a surface functionalized with poly(dT).

[0267] Example 122 According to any one of Examples 111 to 121, each of the plurality of particles has a predetermined particle size.

[0268] Example 123 According to the method described in Example 122, the predetermined particle size is approximately 50 µm.

[0269] Example 124 According to any one of Examples 111 to 123, each of the plurality of particles is porous.

[0270] Example 125 The method according to any one of Examples 111 to 124 further includes deforming the elastic layer into the second chamber portion, thereby driving the at least one liquid out of the second chamber portion through the filter and into the at least one fluid channel.

[0271] Example 126 According to the method of Example 125, the filter allows the at least one liquid to flow through it during the deformation step, and the filter prevents the plurality of particles from flowing through it during the deformation step.

[0272] Example 127 According to any one of Examples 111 to 126, the second layer further defines a particle inlet channel in fluid communication with the second chamber portion, and the method further includes delivering the plurality of particles to the second chamber portion via the particle inlet channel.

[0273] Example 128 According to any one of Examples 111 to 127, the first layer further defines a third chamber portion configured to receive pressurized gas, and the second layer further defines: (i) a fourth chamber portion located below the third chamber portion, the fourth chamber portion being configured to receive the at least one liquid and the plurality of particles, and (ii) at least one bridging channel extending between the second chamber portion and the fourth chamber portion; the method further includes conveying the at least one liquid and the plurality of particles between the second chamber portion and the fourth chamber portion via the at least one bridging channel.

[0274] Example 129 According to any one of Examples 111 to 128, the second layer further defines a particle outlet channel in fluid communication with the second chamber portion, and the method further includes removing the plurality of particles from the second chamber portion via the particle outlet channel.

[0275] Example 130 A method of using an apparatus, the apparatus comprising: (a) a first layer defining a first chamber portion configured to receive pressurized gas; (b) a second layer defining: (i) a second chamber portion located below the first chamber portion, the second chamber portion being configured to receive at least one liquid and a plurality of particles; and (ii) a fluid channel in fluid communication with the second chamber portion; (c) an elastic layer disposed between the first layer and the second layer; and (d) at least one filter channel located between the second chamber portion and the fluid channel, the at least one filter channel being defined by at least one of the second layer or the elastic layer; the method comprising treating the at least one liquid with the plurality of particles within the second chamber portion.

[0276] Example 131 According to the method of Example 130, the at least one liquid contains an RNA therapeutic agent.

[0277] Example 132 According to any one of Examples 130 to 131, the plurality of particles comprises a plurality of beads.

[0278] Example 133 According to any one of Examples 130 to 132, the plurality of particles comprises a plurality of purified particles, and the step of processing the at least one liquid includes purifying the at least one liquid using the plurality of purified particles.

[0279] Example 134 According to the method described in Example 133, each of the plurality of purified particles is configured to separate mRNA from one or more components of the transcription reaction process.

[0280] Example 135 According to the method of Example 134, the one or more components include at least one of plasmid DNA or an enzyme.

[0281] Example 136 According to any one of Examples 134 to 135, each of the plurality of purified particles is configured to separate mRNA from the components of the transcription reaction process by selectively capturing mRNA via a polyadenylate (polyA) tail using a salt and water purification step.

[0282] Example 137 According to any one of Examples 133 to 136, each of the plurality of purified particles includes a rigid polymer resin support matrix.

[0283] Example 138 According to the method of Example 137, the rigid polymer resin support matrix comprises crosslinked poly(styrene-divinylbenzene).

[0284] Example 139 According to the method of any one of Examples 133 to 138, each of the plurality of purified particles has a multi-hydroxyl surface coating.

[0285] Example 140 According to any one of Examples 133 to 139, each of the plurality of purified particles has a surface functionalized with poly(dT).

[0286] Example 141 According to the method of any one of Examples 130 to 140, each of the plurality of particles has a predetermined particle size.

[0287] Example 142 According to the method described in Example 141, the predetermined particle size is approximately 50 µm.

[0288] Example 143 According to any one of Examples 130 to 142, each of the plurality of particles is porous.

[0289] Example 144 The method according to any one of Examples 130 to 143 further includes deforming the elastic layer into the second chamber portion, thereby driving the at least one liquid out of the second chamber portion through the at least one filter channel and into the fluid channel.

[0290] Example 145 According to the method of Example 144, the at least one filter channel allows the at least one liquid to flow through it during the deformation step, and the at least one filter channel prevents the plurality of particles from flowing through it during the deformation step.

[0291] Example 146 According to any one of Examples 130 to 145, the second layer further defines a particle inlet channel in fluid communication with the second chamber portion, and the method further includes delivering the plurality of particles to the second chamber portion via the particle inlet channel.

[0292] Example 147 According to any one of Examples 130 to 146, the first layer further defines a third chamber portion configured to receive pressurized gas, and the second layer further defines: (i) a fourth chamber portion located below the third chamber portion, the fourth chamber portion being configured to receive the at least one liquid and the plurality of particles, and (ii) at least one bridging channel extending between the second chamber portion and the fourth chamber portion; the method further includes conveying the at least one liquid and the plurality of particles between the second chamber portion and the fourth chamber portion via the at least one bridging channel.

[0293] Example 148 According to any one of Examples 130 to 147, the second layer further defines a particle outlet channel in fluid communication with the second chamber portion, and the method further includes removing the plurality of particles from the second chamber portion via the particle outlet channel.

[0294] Example 149 A method of using an apparatus, the apparatus comprising: (a) a first layer defining a first chamber portion configured to receive pressurized gas; (b) a second layer defining: (i) a second chamber portion located below the first chamber portion, the second chamber portion having sidewalls and configured to receive at least one liquid and a plurality of particles; (ii) a fluid channel in fluid communication with the second chamber portion; and (iii) a plurality of sidewall channels extending along the sidewalls; (c) an elastic layer disposed between the first layer and the second layer; and (d) a filter between the second chamber portion and the at least one fluid channel, the filter being defined by at least one of the second layer or the elastic layer, the plurality of sidewall channels extending along the sidewalls to the filter; the method comprising treating the at least one liquid with the plurality of particles within the second chamber portion.

[0295] Example 150 According to the method of Example 149, the at least one liquid contains an RNA therapeutic agent.

[0296] Example 151 According to any one of Examples 149 to 150, the plurality of particles comprises a plurality of beads.

[0297] Example 152 According to any one of Examples 149 to 151, the plurality of particles comprises a plurality of purified particles, and the step of processing the at least one liquid includes purifying the at least one liquid using the plurality of purified particles.

[0298] Example 153 According to the method described in Example 152, each of the plurality of purified particles is configured to isolate mRNA from components of the transcription reaction process.

[0299] Example 154 According to the method of Example 153, the component includes at least one of plasmid DNA or an enzyme.

[0300] Example 155 According to any one of Examples 153 to 154, each of the plurality of purified particles is configured to separate mRNA from the components of the transcription reaction process by selectively capturing mRNA via a polyadenylate (polyA) tail using a salt and water purification step.

[0301] Example 156 According to any one of Examples 152 to 155, each of the plurality of purified particles comprises a rigid polymer resin support matrix.

[0302] Example 157 According to the method of Example 156, the rigid polymer resin support matrix comprises crosslinked poly(styrene-divinylbenzene).

[0303] Example 158 According to the method of any one of Examples 152 to 157, each of the plurality of purified particles has a multi-hydroxyl surface coating.

[0304] Example 159 According to any one of Examples 152 to 158, each of the plurality of purified particles has a surface functionalized with poly(dT).

[0305] Example 160 According to any one of Examples 149 to 159, each of the plurality of particles has a predetermined particle size.

[0306] Example 161 According to the method described in Example 160, the predetermined particle size is about 50 µm.

[0307] Example 162 According to any one of Examples 149 to 161, each of the plurality of particles is porous.

[0308] Example 163 The method according to any one of Examples 149 to 162 further includes deforming the elastic layer into the second chamber portion to expel the at least one liquid from the second chamber portion via the plurality of sidewall channels, through the filter, and into the fluid channel.

[0309] Example 164 According to the method of Example 163, the filter allows the at least one liquid to flow through it during the deformation step, and the filter prevents the plurality of particles from flowing through it during the deformation step.

[0310] Example 165 According to any one of Examples 149 to 164, the second layer further defines a particle inlet channel in fluid communication with the second chamber portion, and the method further includes delivering the plurality of particles to the second chamber portion via the particle inlet channel.

[0311] Example 166 According to any one of Examples 149 to 165, the first layer further defines a third chamber portion configured to receive pressurized gas, and the second layer further defines: (i) a fourth chamber portion located below the third chamber portion, the fourth chamber portion being configured to receive the at least one liquid and the plurality of particles, and (ii) at least one bridging channel extending between the second chamber portion and the fourth chamber portion; the method further includes conveying the at least one liquid and the plurality of particles between the second chamber portion and the fourth chamber portion via the at least one bridging channel.

[0312] Example 167 According to any one of Examples 149 to 166, the second layer further defines a particle outlet channel in fluid communication with the second chamber portion, and the method further includes removing the plurality of particles from the second chamber portion via the particle outlet channel.

[0313] Example 168 According to any one of Examples 2-25, each of the plurality of filter channels has at least one cross-sectional dimension substantially greater than about 50 µm.

[0314] Example 169 According to the fluid device described in Example 168, the at least one cross-sectional dimension is substantially less than about 250 µm.

[0315] Example 170 According to any one of Examples 168-169, the at least one cross-sectional dimension includes at least one of width or depth.

[0316] Example 171 According to the fluid device described in Example 170, the at least one cross-sectional dimension includes both width and depth.

[0317] Example 172 According to any one of Examples 170-171, the fluid device has a depth greater than or equal to about 100 µm.

[0318] Example 173 According to any one of Examples 170-172, the width ranges from about 100µm to about 400µm.

[0319] Example 174 According to the fluid device described in Example 173, the width is approximately 200 µm.

[0320] Example 175 According to any one of Examples 1-25 or 168-174, the second layer defines at least one backflush passage in fluid communication with the at least one fluid outlet passage, the at least one backflush passage being configured to guide at least one backflush fluid through the filter and into the second chamber portion.

[0321] Example 176 According to any one of Examples 26-29, the fluid apparatus, the second layer defines at least one backflush channel in fluid communication with the plurality of filter channels, the at least one backflush channel being configured to guide at least one backflush fluid through the plurality of filter channels and into the second chamber portion.

[0322] Example 177 According to any one of Examples 31 to 49, the second layer further defines at least one backflush passage in fluid communication with the at least one fluid outlet passage, and the method further includes guiding at least one backflush fluid from the at least one backflush passage through the filter and into the second chamber portion.

[0323] Example 178 According to any one of Examples 50-68, the second layer defines at least one backflush channel in fluid communication with the plurality of filter channels, and the method further includes guiding at least one backflush fluid from the at least one backflush channel through the plurality of filter channels and into the second chamber portion.

[0324] Example 179 According to any one of Examples 69 to 87, the second layer defines at least one backflush passage in fluid communication with the at least one fluid outlet passage, and the method further includes guiding at least one backflush fluid from the at least one backflush passage through the filter and into the second chamber portion.

[0325] Example 180 According to any one of Examples 89-105, the at least one filter channel has at least one cross-sectional dimension substantially greater than about 50 µm.

[0326] Example 181 According to the fluid device described in Example 180, the at least one cross-sectional dimension is substantially less than about 250 µm.

[0327] Example 182 According to any one of Examples 180-181, the at least one cross-sectional dimension includes at least one of width or depth.

[0328] Example 183 According to the fluid device described in Example 182, the at least one cross-sectional dimension includes both width and depth.

[0329] Example 184 According to any one of Examples 182-183, the fluid device has a depth greater than or equal to about 100 µm.

[0330] Example 185 According to any one of Examples 182-184, the width ranges from about 100µm to about 400µm.

[0331] Example 186 According to the fluid device described in Example 185, the width is approximately 200 µm.

[0332] Example 187 According to any one of Examples 88-105 or 180-186, the second layer defines at least one backflush passage in fluid communication with the at least one fluid outlet passage, the at least one backflush passage being configured to guide at least one backflush fluid through the filter and into the second chamber portion.

[0333] Example 188 According to any one of Examples 106-109, in the fluid apparatus, the second layer defines at least one backflush passage in fluid communication with the at least one filter passage, the at least one backflush passage being configured to guide at least one backflush fluid through the at least one filter passage and into the second chamber portion.

[0334] Example 189 According to the fluid apparatus of Example 110, the second layer defines at least one backflush channel in fluid communication with the at least one fluid channel, the at least one backflush channel being configured to guide at least one backflush fluid through the filter and into the second chamber portion.

[0335] Example 190 According to any one of Examples 111-129, the second layer defines at least one backflush channel in fluid communication with the at least one fluid channel, and the method further includes guiding at least one backflush fluid from the at least one backflush channel through the filter and into the second chamber portion.

[0336] Example 191 According to any one of Examples 130-148, the second layer defines at least one backflush channel in fluid communication with the at least one filter channel, and the method further includes guiding at least one backflush fluid from the at least one backflush channel through the at least one filter channel and into the second chamber portion.

[0337] Example 192 According to any one of Examples 149-167, the second layer defines at least one backflush channel in fluid communication with the at least one fluid channel, and the method further includes guiding at least one backflush fluid from the at least one backflush channel through the filter and into the second chamber portion.

[0338] VII. Other matters The foregoing description is provided to enable those skilled in the art to practice the various configurations described herein. While the subject matter has been specifically described with reference to various accompanying drawings and configurations, it should be understood that these are for illustrative purposes only and should not be considered as limiting the scope of the subject matter.

[0339] There may be many other ways to implement the subject matter technology. The various functions and elements described herein can be divided differently from those shown without departing from the scope of the subject matter technology. Various modifications to these embodiments may be readily apparent to those skilled in the art, and the general principles defined herein can be applied to other embodiments. Therefore, many changes and modifications can be made to the subject matter technology by those skilled in the art without departing from its scope. For example, different numbers of given modules or units may be used, one or more different types of given modules or units may be used, given modules or units may be added, or given modules or units may be omitted.

[0340] Some versions of the examples described herein can be implemented using a processor, which may be part of a computer system and communicate with multiple peripheral devices via a bus subsystem. Versions of the examples described herein implemented using a computer system can be implemented using a general-purpose computer programmed to perform the methods described herein. Alternatively, versions of the examples described herein implemented using a computer system can be implemented using a special-purpose computer with hardware configurations arranged to perform the methods described herein. Versions of the examples described herein can also be implemented using a combination of at least one general-purpose computer and at least one special-purpose computer.

[0341] In versions implemented using computer systems, each processor may include a central processing unit (CPU), microprocessor, application-specific integrated circuit (ASIC), other types of hardware components, and combinations thereof. A computer system may include more than one type of processor. Peripheral devices of a computer system may include a storage subsystem, including, for example, memory devices and file storage subsystems, user interface input devices, user interface output devices, and a network interface subsystem. Input and output devices allow users to interact with the computer system. The network interface subsystem provides an interface to external networks, including interfaces to corresponding interface devices in other computer systems. User interface input devices may include keyboards; pointing devices, such as mice, trackballs, touchpads, or graphics tablets; scanners; touchscreens integrated into a display; audio input devices, such as speech recognition systems and microphones; and other types of input devices. Generally, the term "input device" is intended to encompass all possible types of devices and methods for inputting information into a computer system.

[0342] In versions implemented using computer systems, user interface output devices may include a display subsystem, a printer, a fax machine, or a non-visual display such as an audio output device. The display subsystem may include a cathode ray tube (CRT), a flat panel device such as a liquid crystal display (LCD), a projection device, or some other mechanism for creating a visible image. The display subsystem may also provide a non-visual display, such as an audio output device. Generally, the term "output device" is intended to encompass all possible types of devices and methods for outputting information from a computer system to a user or another machine or computer system.

[0343] In versions implemented using computer systems, the storage subsystem may store programming and data structures that provide the functionality of some or all of the modules and methods described herein. These software modules can typically be executed by the computer system's processor alone or in conjunction with other processors. The memory used in the storage subsystem may include multiple memories, including main random access memory (RAM) for storing instructions and data during program execution and read-only memory (ROM) for storing fixed instructions. The file storage subsystem may provide permanent storage for program and data files and may include hard disk drives, floppy disk drives along with associated removable media, CD-ROM drives, optical drives, or removable media cartridges. Modules implementing the functionality of a particular implementation may be stored in the storage subsystem or in a file storage subsystem on another machine accessible to the processor.

[0344] In versions implemented using computer systems, the computer system itself can be of different types, including personal computers, portable computers, workstations, computer terminals, network computers, televisions, mainframes, server farms, a widely distributed group of loosely networked computers, or any other data processing system or user equipment. Due to the constantly evolving nature of computers and networks, the examples of computer systems described herein are intended only as specific examples for illustrating the disclosed techniques. Many other configurations of computer systems with more or fewer components than those described herein are possible.

[0345] As an article of manufacture rather than a method, a non-transitory computer-readable medium (CRM) may be loaded with processor-executable program instructions. When executed, the program instructions implement one or more of the computer-implemented methods described above. Alternatively, the program instructions may be loaded onto a non-transitory CRM and, when combined with suitable hardware, become one or more components of a computer-implemented system practicing the disclosed methods.

[0346] Underlined and / or italicized headings and subheadings are used for convenience only, do not limit the subject matter, and are mentioned without regard to the interpretation of the subject matter description. All structural and functional equivalents of elements of the various embodiments described throughout this disclosure that are known or will be known by one of ordinary skill in the art are expressly incorporated herein by reference and are intended to be included by the subject matter. Furthermore, the disclosure herein is not intended to be made public, whether or not such disclosure is expressly stated in the foregoing description.

[0347] It should be recognized that all combinations of the foregoing concepts and other concepts discussed in more detail below (assuming that such concepts are not inconsistent with each other) are contemplated as part of the inventive subject matter disclosed herein. In particular, all combinations of the claimed subject matter appearing at the end of this disclosure are contemplated as part of the inventive subject matter disclosed herein.

Claims

1. A fluid device, comprising: (a) A first layer, the first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas; (b) The second layer, which is defined as follows: (i) A second chamber portion located below the first chamber portion, the second chamber portion being configured to receive at least one liquid and a plurality of particles. (ii) at least one fluid outlet passage in fluid communication with the second chamber portion, and (iii) a filter located between the second chamber portion and the at least one fluid outlet channel, the filter being formed in the surface of the second layer; and (c) An elastic layer disposed between the first layer and the second layer, the elastic layer being deformable into the second chamber portion to drive the at least one liquid out of the second chamber portion through the filter and into the at least one fluid outlet channel, the filter being configured to allow the at least one liquid to flow through it and to prevent the plurality of particles from flowing through it.

2. The fluid device according to claim 1, wherein the filter comprises a plurality of filter channels.

3. The fluid apparatus of claim 2, wherein each of the plurality of filter channels is shallower than the at least one fluid outlet channel.

4. The fluid apparatus according to any one of claims 2 to 3, wherein each of the plurality of filter channels has a depth substantially less than about 50 µm.

5. The fluid device according to claim 4, wherein each of the plurality of filter channels has a depth of approximately 25 µm.

6. The fluid apparatus of claim 4, wherein the depth of each of the plurality of filter channels ranges from about 10 µm to about 23 µm.

7. The fluid apparatus according to any one of claims 2 to 6, wherein each of the plurality of filter channels extends between a corresponding inner end located near the second chamber portion and a corresponding outer end located away from the second chamber portion.

8. The fluid apparatus of claim 7, wherein the filter further comprises a filter inlet extending between the inner ends of the plurality of filter channels.

9. The fluid apparatus according to any one of claims 7 to 8, wherein the filter further comprises a filter outlet extending between the outer ends of the plurality of filter channels.

10. The fluid apparatus according to any one of claims 1 to 9, wherein the second chamber portion has a sidewall, and the second layer defines at least one sidewall passage extending along the sidewall.

11. The fluid device of claim 10, wherein the at least one sidewall channel extends along the sidewall to the filter.

12. The fluid device of claim 11, wherein the at least one sidewall channel comprises a plurality of sidewall channels extending along the sidewall to the filter.

13. The fluid apparatus according to any one of claims 10 to 12, wherein the second chamber portion has a base plate, and the second layer defines a base plate channel extending along the base plate.

14. The fluid device of claim 13, wherein the at least one sidewall channel extends along the sidewall to the bottom plate channel.

15. The fluid device according to any one of claims 10 to 14, wherein the at least one sidewall channel has a width of about 500 µm.

16. The fluid device according to any one of claims 10 to 15, wherein the at least one sidewall channel has a depth of about 100 µm.

17. The fluid apparatus according to any one of claims 1 to 16, wherein the second layer further defines a particle inlet channel in fluid communication with the second chamber portion, the particle inlet channel being configured to deliver the plurality of particles to the second chamber portion.

18. The fluid apparatus of claim 17, wherein at least one cross-sectional dimension of the particle inlet channel is larger than the corresponding cross-sectional dimension of the at least one fluid outlet channel.

19. The fluid device according to any one of claims 17 to 18, wherein the particle inlet channel has a width of about 500 µm.

20. The fluid device according to any one of claims 17 to 19, wherein the particle inlet channel has a depth of about 250 µm.

21. The fluid apparatus according to any one of claims 1 to 20, wherein the first layer further defines a third chamber portion configured to receive pressurized gas, and the second layer further defines: (i) a fourth chamber located below the third chamber portion, the fourth chamber portion being configured to receive the at least one liquid and the plurality of particles, and (ii) at least one bridging channel extending between the second chamber portion and the fourth chamber portion, the at least one bridging channel being configured to provide a path for conveying the at least one liquid and the plurality of particles between the second chamber portion and the fourth chamber portion.

22. The fluid device of claim 21, wherein the at least one bridging channel has a width of about 500 µm.

23. The fluid device according to any one of claims 21 to 22, wherein the at least one bridging channel has a depth of about 500 µm.

24. The fluid apparatus according to any one of claims 1 to 23, wherein the second layer further defines a particle outlet channel in fluid communication with the second chamber portion, the particle outlet channel being configured to remove the plurality of particles from the second chamber portion.

25. The fluid apparatus of claim 24, wherein at least one cross-sectional dimension of the particle outlet channel is larger than the corresponding cross-sectional dimension of the at least one fluid outlet channel.

26. A fluid device, comprising: (a) A first layer, the first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas; (b) The second layer, which is defined as follows: (i) A second chamber portion located below the first chamber portion, the second chamber portion being configured to receive at least one liquid and a plurality of particles. (ii) a fluid outlet passage, which is in partial fluid communication with the second chamber, and (iii) a plurality of filter channels located between the second chamber portion and the fluid outlet channel; and (c) An elastic layer disposed between the first layer and the second layer, the elastic layer being deformable into the second chamber portion to drive the at least one liquid out of the second chamber portion through the plurality of filter channels and into the fluid outlet channel, the plurality of filter channels being configured to allow the at least one liquid to flow through therethrough and the plurality of filter channels being configured to prevent the plurality of particles from flowing through therethrough.

27. The fluid apparatus of claim 26, wherein each of the plurality of filter channels extends between a corresponding inner end located near the second chamber portion and a corresponding outer end located away from the second chamber portion.

28. The fluid apparatus of claim 27, wherein the second layer further defines a filter inlet extending between the inner ends of the plurality of filter channels.

29. The fluid device according to any one of claims 26 to 27, wherein the second layer further defines a filter outlet extending between the outer ends of the plurality of filter channels.

30. A fluid device, comprising: (a) A first layer, the first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas; (b) The second layer, which is defined as follows: (i) A second chamber portion located below the first chamber portion, the second chamber portion having sidewalls, the second chamber portion being configured to receive at least one liquid and a plurality of particles. (ii) A fluid outlet channel in partial fluid communication with the second chamber. (iii) A filter, located between the second chamber portion and the at least one fluid outlet channel, the filter being formed in the surface of the second layer, and (iv) A plurality of sidewall channels extending along the sidewall to the filter; and (c) An elastic layer disposed between the first layer and the second layer, the elastic layer being deformable into the second chamber portion to expel the at least one liquid from the second chamber portion via the plurality of sidewall channels, through the filter and into the fluid outlet channel, the filter being configured to allow the at least one liquid to flow through it and to prevent the plurality of particles from flowing through it.

31. A method of using an apparatus, the apparatus comprising: (a) A first layer, the first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas; (b) The second layer, which is defined as follows: (i) A second chamber portion located below the first chamber portion, the second chamber portion being configured to receive at least one liquid and a plurality of particles. (ii) at least one fluid outlet passage in fluid communication with the second chamber portion, and (iii) a filter located between the second chamber portion and the at least one fluid outlet passage; and (c) An elastic layer disposed between the first layer and the second layer; The method includes treating the at least one liquid with the plurality of particles within the second chamber portion.

32. The method of claim 31, wherein the at least one liquid comprises an RNA therapeutic agent.

33. The method according to any one of claims 31 to 32, wherein the plurality of particles comprises a plurality of beads.

34. The method according to any one of claims 31 to 33, wherein the plurality of particles comprises a plurality of purified particles, and the step of treating the at least one liquid comprises purifying the at least one liquid using the plurality of purified particles.

35. The method of claim 34, wherein each of the plurality of purified particles is configured to separate mRNA from one or more components of the transcription reaction process.

36. The method of claim 35, wherein the one or more components comprise at least one of plasmid DNA or an enzyme.

37. The method according to any one of claims 35 to 36, wherein each of the plurality of purified particles is configured to separate mRNA from the components of the transcription reaction process by selectively capturing mRNA via a polyadenylate (polyA) tail using a salt and water purification step.

38. The method according to any one of claims 34 to 37, wherein each of the plurality of purified particles comprises a rigid polymer resin support matrix.

39. The method of claim 38, wherein the rigid polymer resin support matrix comprises crosslinked poly(styrene-divinylbenzene).

40. The method according to any one of claims 34 to 39, wherein each of the plurality of purified particles has a multi-hydroxyl surface coating.

41. The method according to any one of claims 34 to 40, wherein each of the plurality of purified particles has a surface functionalized with poly(dT).

42. The method according to any one of claims 31 to 41, wherein each of the plurality of particles has a predetermined particle size.

43. The method according to claim 42, wherein the predetermined particle size is about 50 µm.

44. The method according to any one of claims 31 to 43, wherein each of the plurality of particles is porous.

45. The method according to any one of claims 31 to 44, further comprising deforming the elastic layer into the second chamber portion to drive the at least one liquid out of the second chamber portion through the filter and into the at least one fluid outlet channel.

46. ​​The method of claim 45, wherein the filter allows the at least one liquid to flow through it during the deformation step, and the filter prevents the plurality of particles from flowing through it during the deformation step.

47. The method of any one of claims 31 to 46, wherein the second layer further defines a particle inlet channel in fluid communication with the second chamber portion, and the method further includes delivering the plurality of particles to the second chamber portion via the particle inlet channel.

48. The method according to any one of claims 31 to 47, wherein the first layer further defines a third chamber portion configured to receive pressurized gas, and the second layer further defines: (i) a fourth chamber located below the third chamber portion, the fourth chamber portion being configured to receive the at least one liquid and the plurality of particles, and (ii) at least one bridging channel extending between the second chamber portion and the fourth chamber portion; The method further includes conveying the at least one liquid and the plurality of particles between the second chamber portion and the fourth chamber portion via the at least one bridging channel.

49. The method of any one of claims 31 to 48, wherein the second layer further defines a particle outlet channel in fluid communication with the second chamber portion, and the method further includes removing the plurality of particles from the second chamber portion via the particle outlet channel.

50. A method of using an apparatus, the apparatus comprising: (a) A first layer, the first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas; (b) The second layer, which is defined as follows: (i) A second chamber portion located below the first chamber portion, the second chamber portion being configured to receive at least one liquid and a plurality of particles. (ii) a fluid outlet passage, which is in partial fluid communication with the second chamber, and (iii) a plurality of filter channels located between the second chamber portion and the fluid outlet channel; and (c) An elastic layer disposed between the first layer and the second layer; The method includes treating the at least one liquid with the plurality of particles within the second chamber portion.

51. The method of claim 50, wherein the at least one liquid comprises an RNA therapeutic agent.

52. The method according to any one of claims 50 to 51, wherein the plurality of particles comprises a plurality of beads.

53. The method according to any one of claims 50 to 52, wherein the plurality of particles comprises a plurality of purified particles, and the step of treating the at least one liquid comprises purifying the at least one liquid using the plurality of purified particles.

54. The method of claim 53, wherein each of the plurality of purified particles is configured to separate mRNA from one or more components of the transcription reaction process.

55. The method of claim 54, wherein the one or more components comprise at least one of plasmid DNA or an enzyme.

56. The method according to any one of claims 54 to 55, wherein each of the plurality of purified particles is configured to separate mRNA from the components of the transcription reaction process by selectively capturing mRNA via a polyadenylate (polyA) tail using a salt and water purification step.

57. The method according to any one of claims 53 to 56, wherein each of the plurality of purified particles comprises a rigid polymer resin support matrix.

58. The method of claim 57, wherein the rigid polymer resin support matrix comprises crosslinked poly(styrene-divinylbenzene).

59. The method according to any one of claims 53 to 58, wherein each of the plurality of purified particles has a multi-hydroxyl surface coating.

60. The method according to any one of claims 53 to 59, wherein each of the plurality of purified particles has a surface functionalized with poly(dT).

61. The method according to any one of claims 50 to 60, wherein each of the plurality of particles has a predetermined particle size.

62. The method according to claim 61, wherein the predetermined particle size is about 50 µm.

63. The method according to any one of claims 50 to 62, wherein each of the plurality of particles is porous.

64. The method according to any one of claims 50 to 63, further comprising deforming the elastic layer into the second chamber portion to expel the at least one liquid from the second chamber portion through the plurality of filter channels and into the fluid outlet channel.

65. The method of claim 64, wherein the plurality of filter channels allow the at least one liquid to flow through during the deformation step, and the plurality of filter channels prevent the plurality of particles from flowing through during the deformation step.

66. The method of any one of claims 50 to 65, wherein the second layer further defines a particle inlet channel in fluid communication with the second chamber portion, and the method further includes delivering the plurality of particles to the second chamber portion via the particle inlet channel.

67. The method according to any one of claims 50 to 66, wherein the first layer further defines a third chamber portion configured to receive pressurized gas, and the second layer further defines: (i) a fourth chamber located below the third chamber portion, the fourth chamber portion being configured to receive the at least one liquid and the plurality of particles, and (ii) at least one bridging channel extending between the second chamber portion and the fourth chamber portion; The method further includes conveying the at least one liquid and the plurality of particles between the second chamber portion and the fourth chamber portion via the at least one bridging channel.

68. The method of any one of claims 50 to 67, wherein the second layer further defines a particle outlet channel in fluid communication with the second chamber portion, and the method further includes removing the plurality of particles from the second chamber portion via the particle outlet channel.

69. A method of using an apparatus, the apparatus comprising: (a) A first layer, the first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas; (b) The second layer, which is defined as follows: (i) A second chamber portion located below the first chamber portion, the second chamber portion having sidewalls, the second chamber portion being configured to receive at least one liquid and a plurality of particles. (ii) A fluid outlet channel in partial fluid communication with the second chamber. (iii) A filter, located between the second chamber portion and the at least one fluid outlet passage, and (iv) A plurality of sidewall channels extending along the sidewall to the filter; and (c) An elastic layer disposed between the first layer and the second layer; The method includes treating the at least one liquid with the plurality of particles within the second chamber portion.

70. The method of claim 69, wherein the at least one liquid comprises an RNA therapeutic agent.

71. The method according to any one of claims 69 to 70, wherein the plurality of particles comprises a plurality of beads.

72. The method according to any one of claims 69 to 71, wherein the plurality of particles comprises a plurality of purified particles, and the step of treating the at least one liquid comprises purifying the at least one liquid using the plurality of purified particles.

73. The method of claim 72, wherein each of the plurality of purified particles is configured to isolate mRNA from components of the transcription reaction process.

74. The method of claim 73, wherein the component comprises at least one of plasmid DNA or an enzyme.

75. The method according to any one of claims 73 to 74, wherein each of the plurality of purified particles is configured to separate mRNA from the components of the transcription reaction process by selectively capturing mRNA via a polyadenylate (polyA) tail using a salt and water purification step.

76. The method according to any one of claims 72 to 75, wherein each of the plurality of purified particles comprises a rigid polymer resin support matrix.

77. The method of claim 76, wherein the rigid polymer resin support matrix comprises crosslinked poly(styrene-divinylbenzene).

78. The method according to any one of claims 72 to 77, wherein each of the plurality of purified particles has a multi-hydroxyl surface coating.

79. The method according to any one of claims 72 to 78, wherein each of the plurality of purified particles has a surface functionalized with poly(dT).

80. The method according to any one of claims 69 to 79, wherein each of the plurality of particles has a predetermined particle size.

81. The method according to claim 80, wherein the predetermined particle size is about 50 µm.

82. The method according to any one of claims 69 to 81, wherein each of the plurality of particles is porous.

83. The method according to any one of claims 69 to 82, further comprising deforming the elastic layer into the second chamber portion to expel the at least one liquid from the second chamber portion, through the filter, and into the fluid outlet channel via the plurality of sidewall channels.

84. The method of claim 83, wherein the filter allows the at least one liquid to flow through it during the deformation step, and the filter prevents the plurality of particles from flowing through it during the deformation step.

85. The method of any one of claims 69 to 84, wherein the second layer further defines a particle inlet channel in fluid communication with the second chamber portion, and the method further includes delivering the plurality of particles to the second chamber portion via the particle inlet channel.

86. The method according to any one of claims 69 to 85, wherein the first layer further defines a third chamber portion configured to receive pressurized gas, and the second layer further defines: (i) a fourth chamber located below the third chamber portion, the fourth chamber portion being configured to receive the at least one liquid and the plurality of particles, and (ii) at least one bridging channel extending between the second chamber portion and the fourth chamber portion; The method further includes conveying the at least one liquid and the plurality of particles between the second chamber portion and the fourth chamber portion via the at least one bridging channel.

87. The method of any one of claims 69 to 86, wherein the second layer further defines a particle outlet channel in fluid communication with the second chamber portion, and the method further includes removing the plurality of particles from the second chamber portion via the particle outlet channel.

88. A fluid device, comprising: (a) A first layer, the first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas; (b) The second layer, which is defined as follows: (i) a second chamber portion located below the first chamber portion, the second chamber portion being configured to receive at least one liquid and a plurality of particles, and (ii) at least one fluid passage in fluid communication with the second chamber portion; (c) An elastic layer disposed between the first layer and the second layer, the elastic layer being deformable into the second chamber portion to expel the at least one liquid from the second chamber portion; and (d) A filter located between the second chamber portion and the at least one fluid channel, the filter being formed in the surface of at least one of the second layer or the elastic layer, the filter being configured to allow the at least one liquid to flow through it, and the filter being configured to prevent the plurality of particles from flowing through it.

89. The fluid apparatus of claim 88, wherein the filter comprises at least one filter channel.

90. The fluid apparatus of claim 89, wherein the at least one filter channel is shallower than the at least one fluid channel.

91. The fluid apparatus according to any one of claims 89 to 90, wherein the depth of the at least one filter channel is substantially less than about 50 µm.

92. The fluid device according to claim 91, wherein the at least one filter channel has a depth of about 25 µm.

93. The fluid apparatus of claim 91, wherein the at least one filter channel has a depth of about 10 µm.

94. The fluid device according to any one of claims 88 to 93, wherein the filter is formed in at least one surface of the second layer.

95. The fluid device according to any one of claims 88 to 93, wherein the filter is formed in at least one surface of the elastic layer.

96. The fluid device according to claim 95, wherein the elastic layer comprises an upper membrane and a lower membrane.

97. The fluid device of claim 96, wherein the filter includes a pore extending through the upper membrane.

98. The fluid device of claim 97, wherein the orifice defines a filter channel between the first layer and the lower membrane.

99. The fluid apparatus according to any one of claims 96 to 98, wherein the filter comprises a first groove and a second groove extending through the lower membrane.

100. The fluid apparatus of claim 99, wherein the first tank and the second tank define a filter inlet and a filter outlet, respectively.

101. The fluid apparatus of claim 100, wherein the filter inlet is in partial fluid communication with the second chamber and the filter outlet is in fluid communication with the at least one fluid channel.

102. The fluid device according to any one of claims 96 to 101, wherein the upper membrane has a thickness of about 25 µm.

103. The fluid device according to any one of claims 96 to 101, wherein the upper membrane has a thickness of about 10 µm.

104. The fluid device according to any one of claims 88 to 103, wherein the at least one fluid passage comprises at least one fluid inlet passage.

105. The fluid device according to any one of claims 88 to 104, wherein the at least one fluid passage includes at least one fluid outlet passage.

106. A fluid device, comprising: (a) A first layer, the first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas; (b) The second layer, which is defined as follows: (i) a second chamber portion located below the first chamber portion, the second chamber portion being configured to receive at least one liquid and a plurality of particles, and (ii) A fluid passage in partial fluid communication with the second chamber; (c) An elastic layer disposed between the first layer and the second layer, the elastic layer being deformable into the second chamber portion to expel the at least one liquid from the second chamber portion; and (d) At least one filter channel located between the second chamber portion and the fluid channel, the at least one filter channel being formed in the surface of at least one of the second layer or the elastic layer, the at least one filter channel being configured to allow the at least one liquid to flow through it, and the at least one filter channel being configured to prevent the plurality of particles from flowing through it.

107. The fluid device according to claim 106, wherein the at least one filter channel is formed in at least one surface of the elastic layer.

108. The fluid device according to claim 107, wherein the elastic layer comprises an upper membrane and a lower membrane.

109. The fluid device of claim 108, wherein the at least one filter channel is defined by a pore extending through the upper membrane.

110. A fluid device, comprising: (a) A first layer, the first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas; (b) The second layer, which is defined as follows: (i) A second chamber portion located below the first chamber portion, the second chamber portion having sidewalls, the second chamber portion being configured to receive at least one liquid and a plurality of particles. (ii) A fluid passage in partial fluid communication with the second chamber, and (iii) A plurality of sidewall channels extending along the sidewall; (c) An elastic layer disposed between the first layer and the second layer, the elastic layer being deformable into the second chamber portion to expel the at least one liquid from the second chamber portion via the plurality of sidewall channels; and (d) A filter between the second chamber portion and the at least one fluid channel, the filter being formed in the surface of at least one of the second layer or the elastic layer, the filter being configured to allow the at least one liquid to flow through it, the filter being configured to prevent the plurality of particles from flowing through it, the plurality of sidewall channels extending along the sidewall to the filter.

111. A method of using an apparatus, the apparatus comprising: (a) A first layer, the first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas; (b) The second layer, which is defined as follows: (i) A second chamber portion located below the first chamber portion, the second chamber portion being configured to receive at least one liquid and a plurality of particles. (ii) at least one fluid passage in fluid communication with the second chamber portion; (c) an elastic layer disposed between the first layer and the second layer; and (d) A filter located between the second chamber portion and the at least one fluid channel, the filter being defined by at least one of the second layer or the elastic layer; The method includes treating the at least one liquid with the plurality of particles within the second chamber portion.

112. The method of claim 111, wherein the at least one liquid comprises an RNA therapeutic agent.

113. The method according to any one of claims 111 to 112, wherein the plurality of particles comprises a plurality of beads.

114. The method according to any one of claims 111 to 113, wherein the plurality of particles comprises a plurality of purified particles, and the step of treating the at least one liquid comprises purifying the at least one liquid using the plurality of purified particles.

115. The method of claim 114, wherein each of the plurality of purified particles is configured to separate mRNA from one or more components of the transcription reaction process.

116. The method of claim 115, wherein the one or more components comprise at least one of plasmid DNA or an enzyme.

117. The method according to any one of claims 115 to 116, wherein each of the plurality of purified particles is configured to separate mRNA from the components of the transcription reaction process by selectively capturing mRNA via a polyadenylate (polyA) tail through a salt and water purification step.

118. The method according to any one of claims 114 to 117, wherein each of the plurality of purified particles comprises a rigid polymer resin support matrix.

119. The method of claim 118, wherein the rigid polymer resin support matrix comprises crosslinked poly(styrene-divinylbenzene).

120. The method according to any one of claims 114 to 119, wherein each of the plurality of purified particles has a multi-hydroxyl surface coating.

121. The method according to any one of claims 114 to 120, wherein each of the plurality of purified particles has a surface functionalized with poly(dT).

122. The method according to any one of claims 111 to 121, wherein each of the plurality of particles has a predetermined particle size.

123. The method according to claim 122, wherein the predetermined particle size is about 50 µm.

124. The method according to any one of claims 111 to 123, wherein each of the plurality of particles is porous.

125. The method according to any one of claims 111 to 124, further comprising deforming the elastic layer into the second chamber portion to drive the at least one liquid out of the second chamber portion through the filter and into the at least one fluid channel.

126. The method of claim 125, wherein the filter allows the at least one liquid to flow through it during the deformation step, and the filter prevents the plurality of particles from flowing through it during the deformation step.

127. The method of any one of claims 111 to 126, wherein the second layer further defines a particle inlet channel in fluid communication with the second chamber portion, and the method further includes delivering the plurality of particles to the second chamber portion via the particle inlet channel.

128. The method according to any one of claims 111 to 127, wherein the first layer further defines a third chamber portion configured to receive pressurized gas, and the second layer further defines: (i) a fourth chamber located below the third chamber portion, the fourth chamber portion being configured to receive the at least one liquid and the plurality of particles, and (ii) at least one bridging channel extending between the second chamber portion and the fourth chamber portion; The method further includes conveying the at least one liquid and the plurality of particles between the second chamber portion and the fourth chamber portion via the at least one bridging channel.

129. The method of any one of claims 111 to 128, wherein the second layer further defines a particle outlet channel in fluid communication with the second chamber portion, and the method further includes removing the plurality of particles from the second chamber portion via the particle outlet channel.

130. A method of using an apparatus, the apparatus comprising: (a) A first layer, the first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas; (b) The second layer, which is defined as follows: (i) a second chamber portion located below the first chamber portion, the second chamber portion being configured to receive at least one liquid and a plurality of particles, and (ii) A fluid passage in partial fluid communication with the second chamber; (c) an elastic layer disposed between the first layer and the second layer; and (d) At least one filter channel, the at least one filter channel being located between the second chamber portion and the fluid channel, the at least one filter channel being defined by at least one of the second layer or the elastic layer; The method includes treating the at least one liquid with the plurality of particles within the second chamber portion.

131. The method of claim 130, wherein the at least one liquid comprises an RNA therapeutic agent.

132. The method according to any one of claims 130 to 131, wherein the plurality of particles comprises a plurality of beads.

133. The method according to any one of claims 130 to 132, wherein the plurality of particles comprises a plurality of purified particles, and the step of treating the at least one liquid comprises purifying the at least one liquid using the plurality of purified particles.

134. The method of claim 133, wherein each of the plurality of purified particles is configured to separate mRNA from one or more components of the transcription reaction process.

135. The method of claim 134, wherein the one or more components comprise at least one of plasmid DNA or an enzyme.

136. The method according to any one of claims 134 to 135, wherein each of the plurality of purified particles is configured to separate mRNA from the components of the transcription reaction process by selectively capturing mRNA via a polyadenylate (polyA) tail through a purification step using salt and water.

137. The method according to any one of claims 133 to 136, wherein each of the plurality of purified particles comprises a rigid polymer resin support matrix.

138. The method of claim 137, wherein the rigid polymer resin support matrix comprises crosslinked poly(styrene-divinylbenzene).

139. The method according to any one of claims 133 to 138, wherein each of the plurality of purified particles has a multi-hydroxyl surface coating.

140. The method according to any one of claims 133 to 139, wherein each of the plurality of purified particles has a surface functionalized with poly(dT).

141. The method according to any one of claims 130 to 140, wherein each of the plurality of particles has a predetermined particle size.

142. The method according to claim 141, wherein the predetermined particle size is about 50 µm.

143. The method according to any one of claims 130 to 142, wherein each of the plurality of particles is porous.

144. The method according to any one of claims 130 to 143, further comprising deforming the elastic layer into the second chamber portion to drive the at least one liquid out of the second chamber portion through the at least one filter channel and into the fluid channel.

145. The method of claim 144, wherein the at least one filter channel allows the at least one liquid to flow through it during the deformation step, and the at least one filter channel prevents the plurality of particles from flowing through it during the deformation step.

146. The method of any one of claims 130 to 145, wherein the second layer further defines a particle inlet channel in fluid communication with the second chamber portion, and the method further includes delivering the plurality of particles to the second chamber portion via the particle inlet channel.

147. The method according to any one of claims 130 to 146, wherein the first layer further defines a third chamber portion configured to receive pressurized gas, and the second layer further defines: (i) a fourth chamber located below the third chamber portion, the fourth chamber portion being configured to receive the at least one liquid and the plurality of particles, and (ii) at least one bridging channel extending between the second chamber portion and the fourth chamber portion; The method further includes conveying the at least one liquid and the plurality of particles between the second chamber portion and the fourth chamber portion via the at least one bridging channel.

148. The method of any one of claims 130 to 147, wherein the second layer further defines a particle outlet channel in fluid communication with the second chamber portion, and the method further includes removing the plurality of particles from the second chamber portion via the particle outlet channel.

149. A method of using an apparatus, the apparatus comprising: (a) A first layer, the first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas; (b) The second layer, which is defined as follows: (i) A second chamber portion located below the first chamber portion, the second chamber portion having sidewalls, the second chamber portion being configured to receive at least one liquid and a plurality of particles. (ii) A fluid passage in partial fluid communication with the second chamber, and (iii) A plurality of sidewall channels extending along the sidewall; (c) an elastic layer disposed between the first layer and the second layer; and (d) A filter located between the second chamber portion and the at least one fluid channel, the filter being defined by at least one of the second layer or the elastic layer, the plurality of sidewall channels extending along the sidewall to the filter; The method includes treating the at least one liquid with the plurality of particles within the second chamber portion.

150. The method of claim 149, wherein the at least one liquid comprises an RNA therapeutic agent.

151. The method according to any one of claims 149 to 150, wherein the plurality of particles comprises a plurality of beads.

152. The method according to any one of claims 149 to 151, wherein the plurality of particles comprises a plurality of purified particles, and the step of treating the at least one liquid comprises purifying the at least one liquid using the plurality of purified particles.

153. The method of claim 152, wherein each of the plurality of purified particles is configured to separate mRNA from components of the transcription reaction process.

154. The method of claim 153, wherein the component comprises at least one of plasmid DNA or an enzyme.

155. The method according to any one of claims 153 to 154, wherein each of the plurality of purified particles is configured to separate mRNA from the components of the transcription reaction process by selectively capturing mRNA via a polyadenylate (polyA) tail using a salt and water purification step.

156. The method according to any one of claims 152 to 155, wherein each of the plurality of purified particles comprises a rigid polymer resin support matrix.

157. The method of claim 156, wherein the rigid polymer resin support matrix comprises crosslinked poly(styrene-divinylbenzene).

158. The method according to any one of claims 152 to 157, wherein each of the plurality of purified particles has a multi-hydroxyl surface coating.

159. The method according to any one of claims 152 to 158, wherein each of the plurality of purified particles has a surface functionalized with poly(dT).

160. The method according to any one of claims 149 to 159, wherein each of the plurality of particles has a predetermined particle size.

161. The method of claim 160, wherein the predetermined particle size is about 50 µm.

162. The method according to any one of claims 149 to 161, wherein each of the plurality of particles is porous.

163. The method according to any one of claims 149 to 162, further comprising deforming the elastic layer into the second chamber portion to expel the at least one liquid from the second chamber portion via the plurality of sidewall channels, through the filter, and into the fluid channel.

164. The method of claim 163, wherein the filter allows the at least one liquid to flow through it during the deformation step, and the filter prevents the plurality of particles from flowing through it during the deformation step.

165. The method of any one of claims 149 to 164, wherein the second layer further defines a particle inlet channel in fluid communication with the second chamber portion, and the method further includes delivering the plurality of particles to the second chamber portion via the particle inlet channel.

166. The method according to any one of claims 149 to 165, wherein the first layer further defines a third chamber portion configured to receive pressurized gas, and the second layer further defines: (i) a fourth chamber located below the third chamber portion, the fourth chamber portion being configured to receive the at least one liquid and the plurality of particles, and (ii) at least one bridging channel extending between the second chamber portion and the fourth chamber portion; The method further includes conveying the at least one liquid and the plurality of particles between the second chamber portion and the fourth chamber portion via the at least one bridging channel.

167. The method of any one of claims 149 to 166, wherein the second layer further defines a particle outlet channel in fluid communication with the second chamber portion, and the method further includes removing the plurality of particles from the second chamber portion via the particle outlet channel.

168. The fluid apparatus according to any one of claims 2-25, wherein each of the plurality of filter channels has at least one cross-sectional dimension substantially greater than about 50 µm.

169. The fluid device according to claim 168, wherein the at least one cross-sectional dimension is substantially less than about 250 µm.

170. The fluid apparatus according to any one of claims 168-169, wherein the at least one cross-sectional dimension includes at least one of width or depth.

171. The fluid device according to claim 170, wherein the at least one cross-sectional dimension includes both width and depth.

172. The fluid device according to any one of claims 170-171, wherein the depth is greater than or equal to about 100 µm.

173. The fluid device according to any one of claims 170-172, wherein the width ranges from about 100µm to about 400µm.

174. The fluid device according to claim 173, wherein the width is about 200µm.

175. The fluid apparatus according to any one of claims 1-25 or 168-174, wherein the second layer defines at least one backflush passage in fluid communication with the at least one fluid outlet passage, the at least one backflush passage being configured to guide at least one backflush fluid through the filter and into the second chamber portion.

176. The fluid apparatus according to any one of claims 26-29, wherein the second layer defines at least one backflush passage in fluid communication with the plurality of filter passages, the at least one backflush passage being configured to guide at least one backflush fluid through the plurality of filter passages and into the second chamber portion.

177. The method of any one of claims 31 to 49, wherein the second layer further defines at least one backflush passage in fluid communication with the at least one fluid outlet passage, the method further comprising guiding at least one backflush fluid from the at least one backflush passage through the filter and into the second chamber portion.

178. The method of any one of claims 50-68, wherein the second layer defines at least one backflush channel in fluid communication with the plurality of filter channels, the method further comprising guiding at least one backflush fluid from the at least one backflush channel through the plurality of filter channels and into the second chamber portion.

179. The method of any one of claims 69 to 87, wherein the second layer defines at least one backflush passage in fluid communication with the at least one fluid outlet passage, the method further comprising guiding at least one backflush fluid from the at least one backflush passage through the filter and into the second chamber portion.

180. The fluid device according to any one of claims 89-105, wherein the at least one filter channel has at least one cross-sectional dimension substantially greater than about 50 µm.

181. The fluid device according to claim 180, wherein the at least one cross-sectional dimension is substantially less than about 250 µm.

182. The fluid apparatus according to any one of claims 180-181, wherein the at least one cross-sectional dimension includes at least one of width or depth.

183. The fluid device according to claim 182, wherein the at least one cross-sectional dimension includes both width and depth.

184. The fluid device according to any one of claims 182-183, wherein the depth is greater than or equal to about 100 µm.

185. The fluid device according to any one of claims 182-184, wherein the width ranges from about 100 µm to about 400 µm.

186. The fluid device according to claim 185, wherein the width is about 200 µm.

187. The fluid apparatus according to any one of claims 88-105 or 180-186, wherein the second layer defines at least one backflush passage in fluid communication with the at least one fluid outlet passage, the at least one backflush passage being configured to guide at least one backflush fluid through the filter and into the second chamber portion.

188. The fluid apparatus according to any one of claims 106-109, wherein the second layer defines at least one backflush passage in fluid communication with the at least one filter passage, the at least one backflush passage being configured to guide at least one backflush fluid through the at least one filter passage and into the second chamber portion.

189. The fluid apparatus of claim 110, wherein the second layer defines at least one backflush passage in fluid communication with the at least one fluid passage, the at least one backflush passage being configured to guide at least one backflush fluid through the filter and into the second chamber portion.

190. The method of any one of claims 111-129, wherein the second layer defines at least one backflush channel in fluid communication with the at least one fluid channel, the method further comprising guiding at least one backflush fluid from the at least one backflush channel through the filter and into the second chamber portion.

191. The method of any one of claims 130-148, wherein the second layer defines at least one backflush passage in fluid communication with the at least one filter passage, the method further comprising guiding at least one backflush fluid from the at least one backflush passage through the at least one filter passage and into the second chamber portion.

192. The method of any one of claims 149-167, wherein the second layer defines at least one backflush channel in fluid communication with the at least one fluid channel, the method further comprising guiding at least one backflush fluid from the at least one backflush channel through the filter and into the second chamber portion.

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