Apparatus and method for treating RNA with beads

Abstract

A fluidic apparatus includes a first layer defining a first chamber portion configured to receive pressurized gas. The apparatus also includes a second layer defining a second chamber portion positioned under 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 apparatus further includes an elastic layer disposed between the first layer and the second layer, the elastic layer being deformable into the second chamber portion to thereby drive the liquid out of the second chamber portion through the filter and into the fluid outlet channel. The filter is configured to permit the liquid to flow therethrough, and to prevent the particles from flowing therethrough.

Description

APPARATUS AND METHOD FOR TREATING RNA WITH BEADSPRIORITY

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

[0002] The subject matter discussed in this section should not be assumed to be prior art merely as a result of its mention in this section. Similarly, a problem mentioned in this section or associated with the subject matter provided as background should not be assumed to have been previously recognized in the prior art. The subject matter in this section merely represents different approaches, which in and of themselves may also correspond to implementations of the claimed technology.

[0003] Some currently available technologies for manufacturing and formulating polynucleotide therapeutics (e.g., mRNA therapeutics, etc.) may expose the products to contamination and degradation. Some available centralized production may be too costly, too slow, or susceptible to contamination for use in therapeutic formulations possibly including multiple polynucleotide species.BRIEF DESCRIPTION OF THE DRAWINGS

[0004] The details of one or more implementations are set forth in the accompanying drawings and the description below. Other features, aspects, and advantages will become apparent from the description, the drawings, and the claims, in which:

[0005] FIG. 1 depicts a schematic view of an example of a system including a microfluidic process chip;

[0006] FIG. 2 depicts an exploded perspective view of examples of components of the system of FIG. 1 ;

[0007] FIG. 3 depicts a top plan view of an example of a process chip that may be incorporated into the system of FIG. 1;

[0008] FIG. 4A depicts a cross-sectional side view of the process chip of FIG. 3 in a first state of operation;

[0009] FIG. 4B depicts a cross-sectional side view of the process chip of FIG. 3 in a second state of operation;

[0010] FIG. 4C depicts a cross-sectional side view of the process chip of FIG. 3 in a third state of operation;

[0011] FIG. 4D depicts a cross-sectional side view of the process chip of FIG. 3 in a fourth state of operation;

[0012] FIG. 4E depicts a cross-sectional side view of the process chip of FIG. 3 in a fifth state of operation;

[0013] FIG. 4F depicts a cross-sectional side view of the process chip of FIG. 3 in a sixth state of operation;

[0014] FIG. 5 depicts a top perspective view of another example of a process chip that may be incorporated into the system of FIG. 1;

[0015] FIG. 6 depicts a bottom perspective view of the process chip of FIG. 5;

[0016] FIG. 7 depicts atop plan view of the process chip of FIG. 5, with certain layers being transparent to show internal features;

[0017] FIG. 8 depicts an exploded perspective view of the process chip of FIG. 5;

[0018] FIG. 9 depicts a top perspective view of a first layer of the process chip of FIG. 5;

[0019] FIG. 10 depicts a bottom perspective view of the first layer of FIG. 9;

[0020] FIG. 11 depicts a bottom plan view of the first layer of FIG. 9;

[0021] FIG. 12 depicts a top plan view of an upper membrane of a second layer of the process chip of FIG. 5

[0022] FIG. 13 depicts a top plan view of a lower membrane of the second layer of the process chip of FIG. 5

[0023] FIG. 14 depicts a top perspective view of a third layer of the process chip of FIG. 5;

[0024] FIG. 15 depicts a bottom perspective view of the third layer of FIG. 14;

[0025] FIG. 16 depicts a top plan view of the third layer of FIG. 14;

[0026] FIG. 17 depicts an enlarged top plan view of area 17 of the third layer of FIG. 14 as indicated in FIG. 16;

[0027] FIG. 18 depicts a partial top perspective view of the third layer of FIG. 14, showing a functionalized particle fdter;

[0028] FIG. 19 depicts an enlarged top plan view of area 19 of the process chip of FIG. 5 as indicated in FIG. 7, with certain layers being transparent to show internal features, showing a flow path of functionalized particles through a portion of the process chip;

[0029] FIG. 20 depicts a schematic view of a portion of another example of a process chip that may be incorporated into the system of FIG. 1 ; and

[0030] FIG. 21 depicts a partial perspective view of another example of a process chip that may be incorporated into the system of FIG. 1.DETAILED DESCRIPTION

[0031] In some aspects, apparatuses and methods are disclosed herein for processing therapeutic polynucleotides. In particular, these apparatuses and methods may be closed path apparatuses and methods that are configured to minimize or eliminate manual handling during operation. The closed path apparatuses and methods may provide anearly entirely aseptic environment, and the components may provide a sterile path for processing from initial input (e.g., template) to output (e.g., compounded therapeutic). Material inputs (e.g., nucleotides, and any chemical components) into theapparatus may be sterile: and may be input into the system without requiring virtually any manual interaction.

[0032] The apparatuses and methods described herein may be used to generate therapeutics at rapid cycle times at high degree of reproducibility. The apparatuses described herein may be configured to provide, in a single integrated apparatus, synthesis, purification, dialysis, compounding, and concentration of one or more therapeutic compositions. Alternatively, one or more of these processes may be carried out in two or more apparatuses as described herein. In some scenarios, the therapeutic compositions may include therapeutic polynucleotides, such as, for example, ribonucleic acids or deoxyribonucleic acids. The polynucleotides may include only natural nucleotide units or may include any kind of synthetic, semi-synthetic, or modified nucleotide units. All or some of the processing steps may be performed in an unbroken fluid processing pathway, which may be configured as one or a series of consumable microfluidic path device(s) — in some instances also referred to herein as a process chip or a biochip (though the chip need not necessarily be used in bio-related applications). The process chip in some examples may be removably installed in an instrument that is part of a larger microfluidic system, such as that shown in FIG. 1. The disclosed apparatuses and methods may be used for the synthesis of patient-specific therapeutics, including compounding, at a point of care (e.g., hospital, clinic, pharmacy, etc.).

[0033] I. Terminology7

[0034] Throughout this specification and the claims which follow, unless the context requires otherwise, the word “comprise,” and variations such as “comprises” and “comprising” means various components may be co-jointly employed in the methods and articles (e.g., compositions and apparatuses including device and methods). For example, the term “comprising” will be understood to imply the inclusion of any stated elements or steps but not the exclusion of any other elements or steps. In general, any of the apparatuses and methods described herein should be understood to be inclusive, but all or a sub-set of the components and / or steps may alternatively be exclusive andmay be expressed as ‘‘consisting of’ or alternatively “consisting essentially of’ the various components, steps, sub-components, or sub-steps.

[0035] As used herein, the singular forms “a,” “an,” and “the” are intended to include the plural forms as well, unless the context clearly indicates otherwise. 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 as “ / ”.

[0036] Spatially relative terms, such as “under,” “below,” “lower,” “over,” “upper,” and the like, may be used herein for ease of description to describe one element or feature's relationship to another element(s) or feature(s) as illustrated in the figures. It will be understood that the spatially relative terms are intended to encompass different orientations of the device in use or operation in addition to the orientation depicted in the figures. For example, if a device in the figures is inverted, elements described as “under” or “beneath” other elements or features would then be oriented “over” the other elements or features. Thus, the term “under” may encompass both an orientation of over and under. The device may be otherwise oriented (rotated 90 degrees or at other orientations) and the spatially relative descriptors used herein interpreted accordingly. Similarly, the terms “upwardly,” “downwardly,” “vertical,” “horizontal,” and the like are used herein for the purpose of explanation only unless specifically indicated otherwise.

[0037] When a feature or element is herein referred to as being “on” another feature or element, it may be directly on the other feature or element or intervening features and / or elements may also be present. In contrast, when a feature or element is referred to as being “directly on” another feature or element, there are no intervening features or elements present. When a feature or element is referred to as being “connected,” “attached,” or “coupled” to another feature or element, it may be directly connected, attached, or coupled to the other feature or element or intervening features or elements may be present. In contrast, when a feature or element is referred to as being “directly connected,” “directly attached,” or “directly coupled” to another feature or element, there are no intervening features or elements present. Although described or shown with respect to one embodiment, the features and elements so described or shown mayapply to other embodiments. It will also be appreciated by those skilled in the art that references to a structure or feature that is disposed “adjacent” another feature may have portions that overlap or underlie the adjacent feature.

[0038] As used herein in the specification and claims, including as used in the examples and unless otherwise expressly specified, all numbers may be read as if prefaced by the word “about” or “approximately,” even if the term does not expressly appear. The phrase “about” or “approximately” may be used when describing magnitude and / or position to indicate that the value and / or position described is within a reasonable expected range of values and / or positions. For example, a numeric value may have a value that is ±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 values given herein should also be understood to include about or approximately that value unless the context indicates otherwise. For example, if the value “10” is disclosed, then “about 10” is also disclosed. Any numerical range recited herein is intended to include all sub-ranges subsumed therein.

[0039] It is also understood that when a value is disclosed that “less than or equal to” the value, “greater than or equal to the value,” and possible ranges between values are also disclosed, as appropriately understood by the skilled artisan. For example, if the value “X” is disclosed the “less than or equal to X” as well as “greater than or equal to X” (e.g., where X is a numerical value) is also disclosed. It is also understood that the throughout the application, data is provided in a number of different formats, and that this data, represents endpoints and starting points, and ranges for any combination of the data points. For example, if a particular data point “10” and a particular data point “15” are disclosed, it is understood that greater than, greater than or equal to, less than, less than or equal to, and equal to 10 and 15 are considered disclosed as well as between 10 and 15. It is also understood that each unit between two particular units are also disclosed. For example, if 10 and 15 are disclosed, then 1 1, 12, 13, and 14 are also disclosed.

[0040] Although 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 indicates otherwise. These terms are used to distinguish one feature / element from another feature / element, and unless specifically pointed out, do not denote a certain order. Thus, a first feature / element discussed below could be termed a second feature / element, and similarly, a second feature / element discussed below could be termed a first feature / element without departing from the teachings of the present invention.

[0041] As used herein, the terms “system,” “apparatus,” and “device” may be read as being interchangeable with each other. A system, apparatus, and device may each include a plurality of components having various kinds of structural and / or functional relationships with each other.

[0042] As used herein, “polynucleotide” refers to a nucleic acid molecule containing multiple nucleotides and generally refers both to “oligonucleotides’" (a polynucleotide molecule of 18-25 nucleotides in length) and polynucleotides of 26 or more nucleotides. Aspects of this disclosure include compositions including oligonucleotides having a length of 18-25 nucleotides (e.g., 18-mers, 19-mers, 20-mers, 21-mers, 22-mers, 23- mers. 24-mers, or 25-mers), or medium-length polynucleotides having a length of 26 or more nucleotides (e.g., polynucleotides of 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, about 65, about 70, about 75, about 80, about 85, about 90, about 95, about 100, about 110, about 120, about 130, about 140, about 150, about 160, about 170, about 180, about 190, about 200, about 210, about 220, about 230, about 240. about 250. about 260, about 270, about 280, about 290, or about 300 nucleotides), or long polynucleotides having a length greater than about 300 nucleotides (e.g., polynucleotides of between about 300 to about 400 nucleotides, betw een about 400 to about 500 nucleotides, between about 500 to about 600 nucleotides, between about 600 to about 700 nucleotides, between about 700 to about 800 nucleotides, between about 800 to about 900 nucleotides, between about 900 to about 1000 nucleotides, between about 300 to about 500 nucleotides, between about 300 to about 600 nucleotides, between about 300 to about 700 nucleotides, between about 300 to about 800nucleotides, between about 300 to about 900 nucleotides, or about 1000 nucleotides in length, or even greater than about 1000 nucleotides in length, such as about 1500, about 2000, about 2500, about 5000, or about 7000 nucleotides in length). Where a polynucleotide is double-stranded, its length may be similarly described in terms of base pairs.

[0043] As used herein “amplification” may refer to polynucleotide amplification. Amplification may include any suitable method for amplification of a polynucleotide 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.

[0044] As used herein a “cassette” (e.g., a synthetic in vitro transcription facilitator cassette) refers to a polynucleotide sequence which may include or be operably linked to one or more expression elements such as an enhancer, a promoter, a leader, an intron, a 5' untranslated region (UTR), a 3' UTR, or a transcription termination sequence. In some aspects, a cassette comprises at least a first polynucleotide sequence capable of initiating transcription of an operably linked second polynucleotide sequence (which may comprise a template) and optionally a transcription termination sequence operably linked to the second polynucleotide sequence. The template, as described below, may comprise a sequence of interest, for example, an open reading frame (“ORF”) of interest. The cassette may be provided as a single element or as two or more unlinked elements.

[0045] As used herein, a “template” refers to a nucleic acid sequence that contains a sequence of interest for preparing a therapeutic polynucleotide according to the disclosed methods. Templates may be, but are not limited to, a double stranded DNA (dsDNA), an engineered plasmid construct, a cDNA sequence, or a linear nucleic acid sequence (for example, a linear template generated by PCR or by annealing chemically synthesized oligonucleotides). The template may, in certain aspects, be integrated into a “cassette” as described above.

[0046] As used herein, the term ‘‘sequence of interest” refers to a polynucleotide sequence, the use of which may be deemed desirable for a suitable purpose, in particular, for the manufacture of an mRNA for a therapeutic use, and includes but is not limited to, coding sequences of structural genes, and non-coding regulatory sequences that do not encode and mRNA or protein product.

[0047] As used herein, “in vitro transcription” or “IVT” refer to the process whereby transcription occurs in vitro in a non-cellular system to produce synthetic RNA molecules (e.g., synthetic mRNA) for use in various applications, including for therapeutic delivery to a subject, for example, as a therapeutic polynucleotide, which may be part of, or may be used to form, a therapeutic polynucleotide composition as described below. The therapeutic polynucleotide, (e.g., synthetic RNA molecules (transcription product)) generated may be combined with a delivery vehicle to form a therapeutic polynucleotide composition. Synthetic transcription products include mRNAs, antisense RNA molecules, shRNA, circular RNA molecules, ribozy mes, and the like. An IVT reaction may use a purified linear DNA template comprising a promoter sequence and the sequence of the open reading frame (ORF) of a sequence of interest, ribonucleotide triphosphates or modified ribonucleotide triphosphates, a buffer system that includes DTT and magnesium ions, and a phage RNA polymerase.

[0048] As used herein a ‘'therapeutic polynucleotide” refers to a polynucleotide (e.g., an mRNA) that may be part of a therapeutic polynucleotide composition for delivery to a subject to treat a symptom, disease, or condition in a subject; prevent a symptom, disease, or condition in a subject; or to improve or otherwise modify the subject’s health.

[0049] As used herein a “therapeutic polynucleotide composition” (or “therapeutic composition” for short) may refer to a composition including one or more therapeutic polynucleotides (e.g., mRNA) encapsulated by a deliver}' vehicle, which composition may be administered to a subject in need thereof using any suitable administration routes, such as intratumoral, intramuscular, etc. injection. An example of a therapeutic polynucleotide composition is an mRNA (therapeutic) nanoparticle comprising at leastone mRNA encapsulated by a delivery vehicle molecule. An mRNA vaccine is one example of a therapeutic polynucleotide composition.

[0050] As used herein, ‘"delivery vehicle” refers to any substance that facilitates, at least in part, the in vivo, in vitro, or ex vivo delivery of a polynucleotide (e.g.. therapeutic polynucleotide) to targeted cells or tissues (e.g., tumors, etc ). Referring to something as a delivery vehicle need not exclude the possibility of the delivery vehicle also having therapeutic effects. Some versions of a delivery vehicle may provide additional therapeutic effects. In some versions, a delivery vehicle may be a peptoid molecule, such as an amino-lipidated peptoid molecule, that may be used to at least partially encapsulate mRNA. The term “DV” will also be used herein as a shorthand for “delivery vehicle.”

[0051] As used herein, “joining” refers to methods such as ligation, synthesis, primer extension, annealing, recombination, or hybridization use to couple one component to another.

[0052] As used herein “purifying” refers to physical and / or chemical separation of a component (e.g., particles) of other unwanted components (e.g., contaminating substances, fragments, etc.).

[0053] As used herein, the term “substantially free” as used with respect to a given substance, includes 100% free of a given substance, or which comprises less than about 1.0%, or less than about 0.5%, or less than about 0.1% of the given substance.

[0054] II. Overview of System Including Microfluidic Process Chip

[0055] FIG. 1 depicts examples of various components that may be incorporated into a system (100). System (100) of this example includes a housing (103) enclosing a seating mount (11 ) that may removably hold one or more microfluidic process chips (111). In other words, system (100) includes a chip-receiving component that is configured to removably accommodate a process chip (11 1), where the process chip (111) itself defines one or more microfluidic channels or fluid pathways. Components of system (100) (e.g., within housing (103)) that fluidically interact with process chip (111) may include fluid channels or pathways that are not necessarily considered microfluidic (e.g., with such fluid channels or pathways being larger than themicrofluidic channels or fluid pathways in process chip (111)). In some versions, process chips (111) are provided and utilized as single-use devices, while the rest of system (100) is reusable. Housing (103) may be in the form of a chamber, enclosure, etc., with an opening that may be closed (e.g., via a lid or door, etc.) to thereby seal the interior. Housing (103) may enclose a thermal regulator and / or may be configured to be enclosed in a thermally-regulated environment (e g., a refrigeration unit, etc.). Housing (103) may form an aseptic barrier. In some variations, housing (103) may form a humidified or humidity-controlled environment. In addition, or in the alternative, 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 filtering and air flow management and may promote reagents being kept at a desired temperature through the manufacturing process. In addition, such a cabinet may be equipped with UV lamps for sterilization of process chip (111) and other components of system (100). Other suitable features may be incorporated into a cabinet that houses system (100).

[0056] In some scenarios, the assembly formed by housing (103) and the components of system (100) that are within housing (103), without process chip (111), may be considered as being an “instrument.’’ While controller (121) and user interface (123) are shown in FIG. 1 as being outside of housing (103), controller (121) and user interface (123) may in fact be provided in or on housing (103) and may thus also form part of the instrument. As described in greater detail below, this instrument may removably receive process chip (11 1) via a seating mount (1 15). When process chip (111) is seated in seating mount (115), the instrument and process chip (111) cooperate to together form system (100). When process chip (111) is removed from seating mount (115), the portion of system (100) that is left may be regarded as the “instrument.” The instrument, the system (100). and process chip (111) may each be considered an “apparatus.” The term “apparatus” may thus be read to include the instrument by itself, a process chip (111) by itself, the combination of the instrument and process chip (111), some other combination of components of system (100), or some other permutation of system (100) or components thereof.

[0057] Seating mount (115) may be configured to secure process chip (111) using one or more pins or other components configured to hold process chip (111) in a fixed and predefined orientation. Seating mount (115) may thus facilitate process chip (111) being held at an appropriate position and orientation in relation to other components of system (100). In the present example, seating mount (115) is configured to hold process chip (111) in a horizontal orientation, such that process chip (111) is parallel with the ground.

[0058] In some variations, a thermal control (113) may be located adjacent to seating mount (1 15), to modulate the temperature of any process chip (111) mounted in seating mount (115). Thermal control (113) may include a thermoelectric component (e.g., Peltier device, etc.) and / or one or more heat sinks for controlling the temperature of all or a portion of any process chip (111) mounted in seating mount (115). In some variations, more than one thermal control (1 13) may be included, such as to separately regulate the temperature of different ones of one or more regions of process chip (111). Thermal control (113) may include one or more thermal sensors (e.g., thermocouples, etc.) that may be used for feedback control of process chip (111) and / or thermal control (113).

[0059] As shown in FIG. 1. a fluid interface assembly (109) couples process chip (11 1) with a pressure source (117), thereby providing one or more paths for fluid (e.g., gas) at a positive or negative pressure to be communicated from pressure source (117) to one or more interior regions of process chip (111) as will be described in greater detail below. While only one pressure source (117) is shown, system (100) may include two or more pressure sources (117). In some scenarios, pressure may be generated by one or more sources other than pressure source (117). For instance, one or more vials or other fluid sources within reagent storage frame (107) may be pressurized. In addition, or in the alternative, reactions and / or other processes carried out on process chip (111) may generate additional fluid pressure. In the present example, fluid interface assembly (109) also couples process chip (1 11) with a reagent storage frame (107), thereby providing one or more paths for liquid reagents, etc., to be communicated from reagent storage frame (107) to one or more interior regions of process chip (111) as will be described in greater detail below.

[0060] In some versions, pressurized fluid (e.g., gas) from at least one pressure source (117) reaches fluid interface assembly (109) via reagent storage frame (107). such that reagent storage frame (107) includes one or more components interposed in the fluid path between pressure source (117) and fluid interface assembly (109). In some versions, one or more pressure sources (117) are directly coupled with fluid interface assembly, such that the positively pressurized fluid (e.g., positively pressurized gas) or negatively pressurized fluid (e.g., suction or other negatively pressunzed gas) bypasses reagent storage frame (107) to reach fluid interface assembly (109). Regardless of whether the fluid interface assembly (109) is interposed in the fluid path between pressure source (117) and fluid interface assembly (109), fluid interface assembly (109) may be removably coupled to the rest of system (100). such that at least a portion of fluid interface assembly (109) may be removed for sterilization between uses. As described in greater detail below, pressure source (117) may selectively pressurize one or more chamber regions on process chip (111). In addition, or in the alternative, pressure source may also selectively pressurize one or more vials or other fluid storage containers held by reagent storage frame (107).

[0061] Reagent storage frame (107) is configured to contain a plurality of fluid sample holders, each of which may hold a fluid vial that is configured to hold a reagent (e.g., nucleotides, solvent, water, etc.) for deliver}' to process chip (111). In some versions, one or more fluid vials or other storage containers in reagent storage frame (107) may be configured to receive a product from the interior of the process chip (111). In addition, or in the alternative, a second process chip (1 11) may receive a product from the interior of a first process chip (111), such that one or more fluids are transferred from one process chip (111) to another process chip (111). In some such scenarios, the first process chip (111) may perform a first dedicated function (e.g., synthesis, etc.) while the second process chip (1 11) performs a second dedicated function (e.g., encapsulation, etc.). Reagent storage frame (107) of the present example includes a plurality of pressure lines and / or a manifold configured to divide one or more pressure sources (117) into a plurality of pressure lines that may be applied to process chip (111). Such pressure lines may be independently or collectively (in subcombinations) controlled.

[0062] Fluid interface assembly (109) may include a plurality of fluid lines and / or pressure lines where each such line includes a biased (e.g., spring-loaded) holder or tip that individually and independently drives each fluid and / or pressure line to process chip (111) when process chip (111) is held in seating mount (115). Any associated tubing (e.g., the fluid lines and / or the pressure lines) may be part of fluid interface assembly (109) and / or may connect to fluid interface assembly (109). In some versions, each fluid line comprises a flexible tubing that connects between reagent storage frame (107), via a connector that couples the vial to the tubing in a locking engagement (e.g., ferrule) and process chip (111). In some versions, the ends of the fluid lines / pressure lines may be configured to seal against process chip (111) (e g., at a corresponding sealing port formed in process chip ( 111)). as described below. In the present example, the connections between pressure source (117) and process chip (1 11), and the connections between vials in reagent storage frame (107) and process chip (111), all form sealed and closed paths that are isolated when process chip (111) is seated in seating mount (115). Such sealed, closed paths may provide protection against contamination when processing therapeutic polynucleotides.

[0063] The vials of reagent storage frame (107) may be pressurized (e.g., > 1 atm pressure, such as 2 atm, 3 atm, 5 atm, or higher). In some versions, the vials may be pressurized by pressure source (117). Negative or positive pressure may thus be applied. For example, the fluid vials may be pressurized to between about 1 and about 20 psig (e.g., 5 psig, 10 psig, etc.). Alternatively, a vacuum (e.g., about -7 psig or about 7 psia) may be applied to draw fluids back into the vials (e.g., vials serving as storage depots) at the end of the process. The fluid vials may be driven at lower pressure than the pneumatic valves as described below, which may prevent or reduce leakage. In some variations, the difference in pressure between the fluid and pneumatic valves may 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 ).

[0064] System (100) of the present example further includes a magnetic field applicator (119), which is configured to create a magnetic field at a region of the process chip (111). Magnetic field applicator (119) may include a movable head that is operable to move the magnetic field to thereby selectively isolate products that are adhered tomagnetic capture beads within vials or other storage containers in reagent storage frame (107).

[0065] System (100) of the present example further includes one or more sensors (105). In some versions, such sensors (105) include one or more cameras and / or other kinds of optical sensors. Such sensors (105) may sense one or more of a barcode, a fluid level within a fluid vial held within reagent storage frame (107), fluidic movement within a process chip (111) that is mounted within seating mount (115). and / or other optically detectable conditions. In versions where a sensor (105) is used to sense barcodes, such barcodes may be included on vials of reagent storage frame (107), such that sensor (105) may be used to identify vials in reagent storage frame (107). In some versions, a single sensor (105) is positioned and configured to simultaneously view such barcodes on vials in reagent storage frame (107), fluid levels in vials in reagent storage frame (107), fluidic movement within a process chip (111) that is mounted within seating mount (115), and / or other optically detectable conditions. In some other versions, more than one sensor (105) is used to view such conditions. In some such versions, different sensors (105) may be positioned and configured to separately view corresponding optically detectable conditions, such that a sensor (105) may be dedicated to a particular corresponding optically detectable condition.

[0066] In versions where sensors (105) include at least one optical sensor, visual / optical markers may be used to estimate yield. For example, fluorescence may be used to detect process yield or residual material by tagging with fluorophores. In addition, or in the alternative, dynamic light scattering (DLS) may be used to measure particle size distributions within a portion of the process chip (111) (e.g., such as a mixing portion of process chip (111)). In some variations, sensor (105) may provide measurements using one or two optical fibers to convey light (e.g., laser light) into process chip (111); and detect an optical signal coming out of process chip (111). In versions where sensor (105) optically detects process yield or residual material, etc., sensor (105) may be configured to detect visible light, fluorescent light, an ultraviolet (UV) absorbance signal, an infrared (IR) absorbance signal, and / or any other suitable kind of optical feedback.

[0067] In versions where sensors (105) include at least one optical sensor that is configured to capture video images, such sensors (105) may record at least some activity on process chip (111). For example, an entire run for synthesizing and / or processing a material (e.g., a therapeutic RNA) may be recorded by one or more video sensors (105), including a video sensor (105) that may visualize process chip (111) (e.g., from above). Processing on process chip (111) may be visually tracked and this video record may be retained for later quality control and / or processing. Thus, the video record of the processing may be saved, stored, and / or transmitted for subsequent review and / or analysis. In addition, as will be described in greater detail below, the video may be used as a real-time feedback input that may affect processing using at least visually observable conditions captured in the video.

[0068] System (100) of the present example may be controlled by a controller (121). 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 controller (121) (e.g., one or more processors, etc.) is / are embedded within system (100) (e.g.. contained within housing (103)). In addition, or in the alternative, one or more components of controller (121) (e.g.. one or more processors, etc.) may be detachably attached or detachably connected with other components of system (100). Thus, at least a portion of controller (121) may be removable. Moreover, at least a portion of controller (121) may be remote from housing (103) in some versions.

[0069] The control by controller (121) may include activating pressure source (117) to apply pressure through process chip (111) to drive fluidic movement, among other tasks. Controller (121) may be completely or partially outside of housing (103); or completely or partially inside of housing (103). Controller (121) may be configured to receive user inputs via a user interface (123) of system (100); and provide outputs to users via user interface (123). In some versions, controller (121) is fully automated to a point where user inputs are not needed. In some such versions, user interface (123) may provide only outputs to users. User interface (123) may include a monitor, a touchscreen, a keyboard, and / or any other suitable features. Controller (121) maycoordinate processing, including moving one or more fluid(s) onto and on process chip(111), mixing one or more fluids on process chip (111), adding one or more components to process chip (111), metering fluid in process chip (111), regulating the temperature of process chip (11 1), applying a magnetic field (e.g., when using magnetic beads), etc. Controller (121) may receive real-time feedback from sensors (105) and execute control algorithms in accordance with such feedback from sensors (105). Such feedback from sensors (105) may include, but need not be limited to, identification of reagents in vials in reagent storage frame (107), detected fluid levels in vials in reagent storage frame (107), detected movement of fluid in process chip (111), fluorescence of fluorophores in fluid in process chip (111), etc. Controller (121) may include software, firmware and / or hardware. Controller (121) may also communicate with a remote server, e.g., to track operation of the apparatus, to re-order materials (e.g., components such as nucleotides, process chips (1 11), etc ), and / or to download protocols, etc.

[0070] FIG. 2 shows examples of certain forms that may be taken by various components of system (100). In particular, FIG. 2 shows a reagent storage frame (150), a fluid interface assembly (152), a seating mount (154), a thermal control (156), and a process chip (200). Reagent storage frame (150), fluid interface assembly (152), seating mount (154), thermal control (156), and process chip (200) of this example may be configured and operable just like reagent storage frame (107), fluid interface assembly (109), seating mount (115), thermal control (113), and process chip (111), respectively, described above. These components are secured relative to a base (180). A set of rods (1 2) support reagent storage frame (150) over fluid interface assembly (152).

[0071] As shown in FIG. 2, a set of optical sensors (160) are positioned at four respective locations along base (180). Optical sensors (160) may be configured and operable like sensors (105) described above. Optical sensors (160) may include off- the-shelf cameras or any other suitable kinds of optical sensors. Optical sensors (160) are positioned such that fluid vials held within reagent storage frame (150) are within the field of view of one or more of optical sensors (160). In addition, process chip (200) is within the field of view of one or more of optical sensors (160). Each optical sensor (160) is movably secured to base (180) via a corresponding rail (184) (e.g., in a gantryarrangement), such that each optical sensor (160) is configured to translate laterallyalong each corresponding rail (184). A linear actuator (186) is secured to each optical sensor (160) and is thereby operable to drive lateral translation of each optical sensor (160) along the corresponding rail (184). Each actuator (186) may be in the form of a drive belt, a drive chain, a drive cable, or any other suitable kind of structure. Controller (121) may drive operation of actuators (186). Optical sensors (160) may be moved along rails (184) during operation of system (100) in order to facilitate viewing of the appropriate regions of vials in reagent storage frame (150) and / or process chip (200). In some scenarios, optical sensors (160) move in unison along corresponding rails (184). In some other scenarios, optical sensors (160) move independently along corresponding rails (184).

[0072] While optical sensors (160) are shown in FIG. 2 as being mounted to base (180), optical sensors (160) may be positioned elsewhere within system (100), in addition to or as an alternative to being mounted to base (180). For instance, some versions of 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 rails, movable cantilever arms, or other structures that allow such optical sensors (160) to be repositioned during operation of system (100). Optical sensors (160) may be positioned in any other suitable locations. While not shown, system (100) may also include one or more sources of light (e.g., electroluminescent panels, etc.) to provide illumination that aids in optical sensing by optical sensors (160).

[0073] In some versions, one or more mirrors are used to facilitate visualization of components of system (100) by optical sensors (160). Such mirrors may allow optical sensors (160) to view components of system (100) that may not otherwise be within the field of view of sensors (160). Such mirrors may be placed directly adjacent to optical sensors (160). In addition, or in the alternative, such mirrors may be placed adjacent to one or more components of system (100) that are to be viewed by optical sensors (160).

[0074] In use of system (100), an operator may select a protocol to run (e.g.. from a library of preset protocols), or the user may enter a new protocol (or modify an existing protocol), via user interface (123). From the protocol, controller (121) may instruct theoperator which kind of process chip (111) to use, what the contents of vials in reagent storage frame (107) should be, and where to place the vials in reagent storage frame (107). The operator may load process chip (111) into seating mount (1 15); and load the desired reagent vials and export vials into reagent storage frame (107). System (100) may confirm the presence of the desired peripherals, identify process chip (111), and scan identifiers (e.g.. barcodes) for each reagent and product vial in reagent storage frame ( 107), facilitating the vials to match the bill-of-reagents for the selected protocol. After confirming the starting materials and equipment, controller (121) may execute the protocol. During execution, valves and pumps are actuated to deliver reagents as described in greater detail below, reagents are blended, temperature is controlled, and reactions occur, measurements are made, and products are pumped to destination vials in reagent storage frame (107).

[0075] III. Example of Process Chip

[0076] FIGS. 3 and 4A-4F depict the example of a process chip (200) in further detail. In combination with the rest of system (100), process chip (200) may be utilized to provide in-vitro synthesis, purification, concentration, formulation, and / or analysis of therapeutic compositions, including but not limited to therapeutic polynucleotides and therapeutic polynucleotide compositions. As shown in FIG. 3, process chip (200) of this example includes a plurality of fluid ports (220). Each fluid port (220) has an associated fluid channel (222) formed in process chip (200), such that fluid communicated into fluid port (220) will flow through the corresponding fluid channel (222). As described in greater detail below, each fluid port (220) is configured to receive fluid from a corresponding fluid line (206) from fluid interface assembly (109). In the present example, each fluid channel (222) leads to a valve chamber (224), which is operable to selectively prevent or permit fluid from the corresponding fluid channel (222) to be further communicated along process chip (200) as will be described in greater detail below.

[0077] As also shown in FIG. 3. process chip (200) of this example includes a plurality of additional chambers (230, 250. 270) that may be used to serve different purposes during the process of producing the therapeutic composition as describedherein. By way of example only, such additional chambers (230, 250, 270) may be used to provide synthesis, purification, dialysis, compounding, and / or concentration of one or more therapeutic compositions; or to perform any other suitable function(s). Fluid may be communicated from one chamber (230) to another chamber (230) via a fluidic connector (232). In some versions, fluidic connector (232) is operable like a valve between an open and closed state (e.g., similar to valve chamber (224)). In some other versions, fluidic connector (232) remains open throughout the process of making the therapeutic composition. In the present example, chambers (230) are used to provide synthesis of polynucleotides, though chambers (230) may alternatively sen e any other suitable purpose(s).

[0078] In the example shown in FIG. 3, another valve chamber (234) is interposed between one of chambers (230) and one of chambers (250), such that fluid may be selectively communicated from chamber (230) to chamber (250). Chambers (250) are provided in a pair and are coupled with each other such that process chip (200) may communicate the fluid back and forth between chambers (250). While a pair of chambers (250) are provided in the present example, any other suitable number of chambers (250) may be used, including just one chamber (250) or more than two chambers (250). Chambers (250) may be used to provide purification of the fluid and / or may serve any of the other various purposes described herein; and may have any suitable configuration. In versions where a chamber (250) is used for purification, chamber (250) may include a material that is configured to absorb selected moieties from a fluidic mixture in chamber (250). In some such versions, the material may include a cellulose material, which may selectively absorb double-stranded mRNA from a mixture. In some such versions, the cellulose material may be inserted in only- one chamber (250) of a pair of chambers (250), such that upon mixing the fluid from the first chamber (250) of the pair to the second chamber (250). mRNA and / or some other component may be effectively removed from the fluidic mixture, which may then be transferred to another pair of chambers (270) further downstream for further processing or export. Alternatively, chambers (250) may be used for any other suitable purpose.

[0079] Additional valve chambers (252) are interposed between each chamber (250) and a corresponding chamber (270). such that fluid may be selectively communicated from chambers (250) to chambers (270) via valve chambers (252). Chambers (270) are also coupled with each other such that process chip (200) may communicate the fluid back and forth between chambers (270). Chambers (270) may be used to provide mixing of the fluid and / or may serve any of the other various purposes described herein; and may have any suitable configuration.

[0080] As shown in FIG. 3. chambers (270) are also coupled with additional fluid ports (221) via corresponding fluid channels (223) and valve chambers (225). Fluid ports (221), fluid channels (223), and valve chambers (225) may be configured and operable like fluid ports (220), fluid channels (222), and valve chambers (224) described above. In some versions, fluid ports (221) are used to communicate additional fluids to chambers (270). In addition, or in the alternative, fluid ports (221) may be used to communicate fluid from process chip (200) to another device. For instance, fluid from chambers (270) may be communicated via fluid ports (221) directly to another process chip (200), to one or more vials in reagent storage frame (107), or elsewhere.

[0081] Process chip (200) further includes several reservoir chambers (260). In this example, each reservoir chamber (260) is configured to receive and store fluid that is being communicated to or from a corresponding chamber (250, 270). Each reservoir chamber (260) has a corresponding inlet valve chamber (262) and outlet valve chamber (264). Each inlet valve chamber (262) is interposed between reservoir chamber (260) and the corresponding chamber (250, 270) and is thereby operable to permit or prevent the flow of fluid between reservoir chamber (260) and the corresponding chamber (250, 270). Each outlet valve chamber (264) is operable to meter the flow7of fluid between reservoir chamber (260) and a corresponding fluid port (266). In some versions, each fluid port (266) is configured to communicate fluid from a corresponding vial in reagent storage frame (107) to a corresponding reservoir chamber (260). In addition, or in the alternative, each fluid port (266) may be configured to communicate fluid from a corresponding reservoir chamber (260) to a corresponding vial in reagent storage frame (107). In the present example, reservoir chambers (260) are used to provide meteringof fluid communicated to and / or from process chip (200). Alternatively, reservoir chambers (260) may be utilized for any other suitable purposes, including but not limited to pressurizing fluid that is communicated to and / or from process chip (200).

[0082] As also shown in FIG. 3, process chip (200) of this example includes a plurality of pressure ports (240). Each pressure port (240) has an associated pressure channel (244) formed in process chip (200), such that pressurized gas communicated through pressure port (240) will be further communicated through the corresponding pressure channel (244). As described in greater detail below, each pressure port (240) is configured to receive pressurized gas from a corresponding pressure line (208) from fluid interface assembly (109). In the present example, each pressure channel (244) leads to a corresponding chamber (224, 225, 230, 234, 250, 252, 260, 262, 264, 270) to thereby provide valving or peristaltic pumping via such chambers (224. 225, 230, 234, 250, 252, 260, 262, 264, 270) as described in greater detail below.

[0083] Process chip (200) may also include electrical contacts, pins, pin sockets, capacitive coils, inductive coils, or other features that are configured to provide electrical communication with other components of system (100). In the example shown in FIG. 3, process chip (200) includes an electrically active region (212) that includes such electrical communication features. Electrically active region (212) may further include electrical circuits and other electrical components. In some versions, electrically active region (212) may provide communication of power, data, etc. While electrically active region (212) is shown in one particular location on process chip, electrically active region (212) may alternatively be positioned at any other suitable location or locations. In some versions, electrically active region (212) is omitted.

[0084] As shown in FIGS. 4A-4F, process chip (200) further includes a first plate (300), an elastic layer (302), a second plate (304), and a third plate (306). As described in greater detail below, some versions of elastic layer (302) are in the form of a flexible membrane. First plate (300) has an upper surface (210) and a lower surface (310), with lower surface (310) apposing elastic layer (302). Second plate (304) has an upper surface (312) and a lower surface (314), with upper surface (312) apposing elastic layer (302); and with lower surface (314) apposing third plate (306). Elastic layer (302) isthus interposed between first and second plates (300, 304). In the present example, another elastic layer (316) is also interposed between second and third plates (304. 306). though this elastic layer (316) is optional.

[0085] Plates (300. 304, 306) of the present example are substantially translucent to visible light and / or ultraviolet light. By ‘'substantially translucent” is meant that at least 90% (including in some instances 100%) of light is transmitted through the material compared to a translucent material. In some variations, the one or more of plates (300, 304, 306) may comprise materials that are substantially transparent to visible light and / or ultraviolet light. By ‘'substantially transparent” is meant that at least 90% (including in some instances 100%) of light is transmitted through the material compared to a completely transparent material. As another example, one or more of plates (300, 304, 306) may provide transmission of ultraviolet light at a wavelength of approximately 260 nm at a transmission rate ranging from approximately 0.2% to approximately 20%, including from approximately 0.4% to approximately 15%, or including from approximately 0.5% to approximately 10%.

[0086] Plates (300, 304, 306) of the present example are also rigid. In some other versions, one or more of plates (300, 304, 306) are semi-rigid. Plates (300, 304. 306) may comprise glass, plastic, silicone, and / or any other suitable material(s). In some versions, one or more of plates (300, 304, 306) is formed as a lamination of two or more layers of material, such that each plate (300, 304, 306) does not necessarily need to be formed as a single homogenous continuum of material. The material(s) comprising one of plates (300, 304, 306) may also differ from the material(s) comprising other plates (300. 304, 306).

[0087] Elastic layer (302) of the present example is formed as a liquid-impermeable flexible membrane. In some versions, elastic layer (302) is gas-permeable despite being liquid-impermeable. In some such versions, certain regions of elastic layer (302) are treated to be gas-permeable while the non-treated regions of elastic layer (302) are gas- impermeable. As described below, elastic layer (302) may be used to drive fluids across process chip (200) via peristaltic pumping action. As also described below, elastic layer (302) may be used to provide valves at various locations along process chip (200). Insome versions, a single sheet of elastic material spans across the width of process chip (200) to form elastic layer (302). In some other versions, two or more discrete pieces of elastic material are used to form elastic layer (302), with such discrete pieces of elastic material being positioned at different locations across the width of process chip (200). By way of example only, elastic layer (302) may include a membrane comprising poly dimethylsilicone (PDMS) elastomer fdm.

[0088] As best seen in FIGS. 4A-4F, first and second plates (300, 304) cooperate to define a plurality of chambers (320. 322, 324, 326), with elastic layer (302) bisecting each chamber (320, 322, 324, 326) into a corresponding upper chamber region (330) and lower chamber region (332). Chambers (224, 225, 230, 234, 250, 252, 260, 262, 264, 270) shown in FIG. 3 may be configured and operable just like chambers (320, 322, 324. 326) shown in FIGS. 4A-4F. For instance, chamber (320) may be analogous to chamber (264), chamber (322) may be analogous to chamber (260), chamber (324) may be analogous to chamber (262), and chamber (326) may be analogous to chamber (250).

[0089] As shown in FIGS. 4A-4F, fluid port (220) is formed through first plate (300). A corresponding opening (342) is formed through the region of elastic layer (302) underlying fluid port (220). Fluid channel (222) extends from opening (342) to lower chamber region (332) of first chamber (320). As noted above, fluid port (220) is configured to receive a fluid line (206) from fluid interface assembly (109). The distal end of fluid line (206) is configured to seal against the region of elastic layer (302) that is exposed by fluid port (220) and communicate fluid (207) through opening (342). In some versions, a spring or other resilient member provides a resilient bias to fluid line (206), urging the distal end of fluid line (206) against the region of elastic layer (302) that is exposed by fluid port (220) to thereby maintain the seal. Fluid (207) from fluid line (206) reaches lower chamber region (332) of first chamber (320) via fluid channel (222). As described in greater detail below, this fluid (207) may be further communicated from first chamber (320) to other chambers (322, 324, 326) through a peristaltic pumping action that is provided via elastic layer (302). After reaching fourth chamber (326), the fluid (207) may be further communicated to other chambers or other features in process chip (200), may be communicated to a storage vial in reagent storageframe (107), or may be otherwise processed. The path for fluid (207) thus does not necessarily terminate at fourth chamber (326). It should also be understood that any of the other fluid ports (221, 266) shown in FIG. 3 may be configured and operable like fluid port (220) shown in FIGS. 4A-4F.

[0090] Pressure port (240) is formed through first plate (300). A corresponding opening (344) is formed through the region of elastic layer (302) underlying pressure port (240). Pressure channel (244) extends from opening (344) to upper chamber region (330) of first chamber (320). As noted above, pressure port (240) is configured to receive a pressure line (208) from fluid interface assembly (109), to thereby receive pressurized gas from pressure source (117). The distal end of pressure line (208) is configured to seal against the region of elastic layer (302) that is exposed by pressure port (240) and communicate either positively pressurized gas or negatively pressurized gas through opening (344). In some versions, a spring or other resilient member provides a resilient bias to pressure line (208), urging the distal end of pressure line (208) against the region of elastic layer (302) that is exposed by pressure port (240) to thereby maintain the seal. Positively pressurized gas or negatively pressurized gas from pressure line (208) reaches upper chamber region (330) of fourth chamber (326) via pressure channel (244).

[0091] While FIGS. 4A-4F depict just one pressure line (208) being coupled with process chip (200), process chip (200) may have several coupled pressure lines (208), with such pressure lines (208) independently applying positive or negative pressure to corresponding chambers (320, 322, 324, 326) of process chip (200). In some versions, one or more of chambers (320. 322, 324. 326) has its own dedicated pressure line (208) and corresponding pressure channel (244). In addition, or in the alternative, one or more of chambers (320, 322, 324, 326) may share a common pressure line (208), via the same pressure channel (244) or via separate pressure channels (244). While FIGS. 4A-4F depict pressure channel (244) formed through second plate (304), some pressure channels (244) (or regions of pressure channels (244)) may be formed by first plate (300). For instance, some pressure channels (244) (or regions of pressure channels (244)) may be formed between a recess in the lower surface of first plate (300) and the top surface of elastic layer (302).

[0092] IV. Example of Valving and Peristaltic Pumping Driven via Elastic Layer

[0093] As noted above, elastic layer (302) may be operated to drive fluid through process chip (200) through a peristaltic pumping action; and to arrest movement of fluid through process chip (200) by providing a valving action. An example of such operation is illustrated in the sequence depicted through FIGS. 4A-4F. In this example, chambers (320, 324) serve as valve chambers, while chamber (322) serves as a metering chamber. Chamber (326) serves as a working chamber, such that synthesis, purification, dialysis, compounding, concentration, or some other process is performed in chamber (326). This configuration, arrangement, and usage 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.

[0094] FIG. 4A shows process chip (200) in a state where fluid is not yet being communicated to process chip (200); and pressurized gas is not yet being communicated to process chip (200). In FIG. 4B, positively pressurized gas is communicated to upper chamber region (330) of chamber (324), negatively pressurized gas is communicated to upper regions (330) of chambers (320, 322), and fluid (207) is communicated to chambers (320, 322). In this state, the positively pressurized gas deforms the portion of elastic layer (302) in chamber (324) such that elastic layer (302) seats against the surface of lower chamber region (332) of chamber (324). This seating of elastic layer (302) against the surface of lower chamber region (332) of chamber (324) prevents fluid (207) from entering chamber (324), such that chamber (324) is operating like a closed valve in the state shown in FIG. 4B. The negatively pressurized gas in upper chamber regions (330) of chambers (320, 322) causes the corresponding portion of elastic layer (302) in chambers (320, 322) to deform and seat against upper chamber regions (330) of chambers (320, 322). This allows fluid (207) to occupy the full capacity of chambers (320, 322).

[0095] After reaching the state shown in FIG. 4B, positively pressurized gas is communicated to upper chamber region (330) of chamber (320) while the pneumatic state of chambers (322, 324) may remain unchanged. This results in the state shown in FIG. 4C. As shown, the positively pressurized gas deforms the portion of elastic layer(302) in chamber (320) such that elastic layer (302) seats against the surface of lower chamber region (332) of chamber (320). This seating of elastic layer (302) against the surface of lower chamber region (332) of chamber (320) drives the fluid (207) out from chamber (320) and results in chamber (320) operating like a closed valve in the state show n in FIG. 4C. However, the volume of fluid (207) in chamber (322) is unaffected in the state shown in FIG. 4C. Chamber (322) may thus be used to provide metering of fluid (207), such that only a precise, predetermined volume of fluid (207) is communicated further along process chip (200). By way of example only, such metered volumes may be on the order of approximately 10 nL, 20 nL, 25 nL, 50 nL, 75 nL, 100 nL, 1 microliter, 5 microliters, etc.

[0096] Once the appropriate metering volume has been achieved, negatively pressurized gas is communicated to upper chamber regions (330) of chambers (324, 326) while the pneumatic state of chambers (320, 322) may remain unchanged. This results in the state shown in FIG. 4D. As show n, the negatively pressurized gas in upper chamber regions (330) of chambers (324, 326) causes the corresponding portion of elastic layer (302) in chamber (324, 326) to deform and seat against the surface of upper chamber regions (330) of chambers (324, 326). This effectively opens the valve formed by chamber (324) and puts chamber (326) in a state to receive fluid (207). This also produces a negative pressure in chamber (324) that draws fluid (207) from chamber (322) into chamber (324).

[0097] With the valve formed by chamber (324) being in the open state, positively pressurized gas is communicated to upper chamber region (330) of chamber (322) while the pneumatic state of chambers (320. 324, 326) may remain unchanged. This results in the state shown in FIG. 4E. As shown, the positively pressurized gas in upper chamber region (330) of chamber (322) causes the corresponding portion of elastic layer (302) in chamber (322) to deform and seat against the surface of lower chamber region (332) of chamber (322). This deformation of elastic layer (302) drives fluid (207) out of chamber (322). Since the valve formed by chamber (320) is in a closed state and the valve formed by chamber (324) is in an open state, fluid (207) travels from chamber (322) into chamber (324). In the present example, the capacity of chamber(322) is greater than the capacity of chamber (324), such that fluid (207) from chamber (322) overflows from chamber (324) into chamber (326).

[0098] Once fluid (207) has been communicated from chamber (322) to chambers (324. 326). positively pressurized gas is communicated to upper chamber region (330) of chamber (324) while the pneumatic state of chambers (320, 322, 326) may remain unchanged. This results in the state shown in FIG. 4F. As shown, the positively pressurized gas in upper chamber region (330) of chamber (324) causes the corresponding portion of elastic layer (302) in chamber (324) to deform and seat against the surface of lower chamber region (332) of chamber (324). This deformation of elastic layer (302) drives fluid (207) out of chamber (324). Since the deformed portion of elastic layer (302) in chamber (324) is effectively sealing off chamber (324) from chamber (322) (e.g., such that chamber (324) is operating like a valve in a closed state), fluid (207) travels from chamber (324) into chamber (326).

[0099] At the stage shown in FIG. 4F, fluid (207) has been evacuated from chambers (320, 332, 324), and chamber (326) contains the volume of fluid (207) that was precisely metered in chamber (322). Fluid (207) in chamber (326) may be further processed within chamber (326) in accordance with the teachings herein. In addition, or in the alternative, fluid (207) in chamber (326) may be communicated to one or more other chambers in process chip (200), may be communicated to a vial in reagent storage frame (107), or may be otherwise handled. Regardless of what is done with fluid (207) after fluid (207) has reached chamber (326), it should be understood that fluid (207) was communicated along chambers (320, 322, 324), in a sequence, to reach chamber (326) via a peristaltic action created through elastic layer (302) in response to positively pressurized gas or negatively pressurized gas being communicated to upper chamber regions (330) of chambers (320, 322, 324, 326) in a particular sequence. Such peristaltic pumping may have particular advantage for moving fluid that may be viscous or contain suspended particles such as purification or capture beads. Such peristaltic pumping through selective deformation of elastic layer (302) may also be referred to as pneumatic barrier deflection or “pneumodeflection.”

[0100] In some scenarios, it may be desirable to remove air or other gas from one or more fluid pathways in process chip (200). To accomplish this, process chip (200) may include one or more chambers that are configured to provide ventilation of a fluid pathway or otherwise evacuate gas from the fluid pathway. For instance, such ventilation or evacuation may be performed as part of a priming process as fluid is initially introduced to process chip (200). In addition, or in the alternative, such ventilation or evacuation may be performed to relieve gas that is generated in the fluid during the process of forming the therapeutic composition. Such ventilation or gas relief chambers may be referred to as “vacuum caps.” In some versions, at least the region of elastic layer (302) that is positioned in the vacuum cap (if not the entirety of elastic layer (302)) is gas permeable (while still being liquid impermeable). Negatively pressurized gas may be applied to the upper chamber region (330) of the chamber that is being used as a vacuum cap, and this negatively pressurized gas may draw the air or gas from the fluid pathway out through the corresponding region of elastic layer (302). In some versions, the upper chamber region (330) of the chamber that is being used as a vacuum cap includes one or more projections or stand-off features that prevent the corresponding region of elastic layer (302) from fully seating against the surface of the upper chamber region (330) of the chamber that is being used as a vacuum cap. This may further promote evacuation of air or other gas via the vacuum cap.

[0101] V. Example of Process Chip with Bead-Containing Chambers

[0102] In some scenarios, it may be desirable to provide a version of process chip (200) that provides one or more process chambers in which the fluid may be treated with functionalized solid particles (e.g., purification beads, etc.); and that prevents such particles from impeding the flow of the fluid through process chip (200) and / or that prevents such particles from traveling to regions of process chip (200) where the presence of such particles may not be desired. For instance, in a version of process chip (200) that is configured to provide RNA purification with one or more purification chambers, it may be desirable to provide purification beads within those purification chambers to facilitate the purification process. In this regard, it will be appreciated that various types of purification beads may be used to purify and / or isolate RNA (e.g., mRNA) from IVT manufacturing processes. In some cases, such beads may include aresin that effectively separates mRNA from components of the transcription reaction process, such as plasmid DNA, enzymes, and other IVT components, by selectively capturing the mRNA via the poly adenylated (poly A) tail using salt and water purification steps. More particularly, such beads may include 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 may be functionalized with poly(dT). The beads may each have a predetermined particle size (e.g., diameter), such as ranging from about 10 pm to about 200 pm, such as about 50 pm, for example; and / or may each be porous to increase the surface area of each bead and thereby effectively increase the contact area between the beads and the fluid. One example of such beads is POROS Oligo (dT)25 Affinity Resin by Thermo Fisher Scientific of Waltham, Massachusetts. Other examples of particles that may be used for purification include urea granules, which may also each have a predetermined particle size (e.g., diameter), such as ranging from about 10 pm to about 200 pm, such as about 50 pm, for example.

[0103] It may therefore be desirable to provide a variation of process chip (200) with channels and valves that are sized and configured to direct such particles to at least one process chamber, such as at least one purification chamber, and to filter such particles out of the treated fluid to prevent the particles from exiting the at least one process chamber with the treated fluid.

[0104] Also in some scenarios, when elastic layer (302) is pneumatically deformed against a surface of a chamber (320. 322, 324, 326) to drive fluid out of that chamber (320. 322, 324. 326). there may be a tendency for elastic layer (302) to trap pockets of fluid against the floor and / or sidewall of chamber (320, 322, 324, 326), such that not all of the fluid exits chamber (320, 322, 324, 326) when elastic layer (302) is pneumatically deformed to drive fluid out of that chamber (320, 322, 324, 326). It may therefore be desirable to provide a feature in a chamber that prevents an elastic layer from trapping pockets of fluid within a chamber when the elastic layer is pneumatically deformed to drive fluid out of that chamber.

[0105] FIGS. 5-8 show an example of a process chip (400) that may provide at least some, if not all. of the features and functionalities described above. Process chip (400) is similar to process chip (200) described above, except as otherwise described below. In this regard, in combination with the rest of system (100), process chip (400) may be utilized to provide in-vitro synthesis, purification, concentration, formulation (e.g., encapsulation of a therapeutic composition in a delivery vehicle through a mixing process, etc.), and / or analysis of therapeutic compositions and / or templates, including but not limited to therapeutic polynucleotides templates, therapeutic polynucleotides, and / or therapeutic polynucleotide compositions. Alternatively, process chip (400) may be utilized in other processes.

[0106] As shown in FIGS. 5 and 7, process chip (400) of this example includes a plurality of fluid ports (410) and a plurality’ of pressure ports (420). Each fluid port (410) is configured to receive fluid from a corresponding fluid line (206) from fluid interface assembly (109). Each pressure port (420) is configured to receive pressurized gas from a corresponding pressure line (208) from fluid interface assembly (109). As also shown in FIGS. 5 and 7, the plurality of fluid ports (410) of this example includes a pair of purification bead ports (410a, 410b), each of which is configured to receive purification beads or similar particles from a corresponding fluid line (206) and / or to deliver beads or similar particles to the corresponding fluid line (206). For example, first purification bead port (410a) may be configured to receive a slurry containing beads suspended in a liquid solution (e.g., 0. 1 M sodium chloride) from a first fluid line (206), and second purification bead port (410b) may be configured to subsequently deliver the slurry' to a second fluid line (206), as discussed in greater detail below. In some cases, it may be desirable to permanently retain the beads within process chip (400). For example, one or both purification bead ports (410a, 410b) may only be used during the initial manufacture or initial setup of process chip (400) for introducing the beads into process chip (400).

[0107] As shown in FIG. 7, process chip (400) of this example further includes a plurality of process chambers (450, 460, 470a, 470b) that may be used to serve different purposes during the process of producing the therapeutic composition as described herein. By way of example only, process chambers (450, 460, 470a, 470b) may be usedto provide synthesis, purification, dialysis, compounding, and / or concentration of one or more therapeutic compositions; or to perform any other suitable function(s). In the present example, process chambers (450) are configured to be used for IVT reaction, and thus may also be referred to as “reaction chambers;” process chambers (460) are configured to be used for DNA digestion, and thus may also be referred to as “digestion chambers;” and process chambers (470a, 470b) are configured to be used for RNA purification, and thus may also be referred to as "purification chambers.” Process chip (400) may have any other suitable number and / or type(s) of process chambers (450, 460, 470a, 470b). As shown in FIG. 8, process chip (400) of this example further 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 greater detail below.

[0108] In the present example, fluid ports (410) and pressure ports (420) are all positioned on a top region of process chip (400), at least within the frame of reference of FIG. 5. In some other versions, fluid ports (410) are all positioned on a top region of process chip (400) while pressure ports (420) are all positioned on a bottom region of process chip (400). In some other versions, fluid ports (410) are all positioned on a bottom region of process chip (400) while pressure ports (420) are all positioned on a top region of process chip (400). As yet another variation, some fluid ports (410) may be positioned on a top region of process chip (400) while other fluid ports (410) are positioned on a bottom region of process chip (400). Similarly, some pressure ports (420) may be positioned on atop region of process chip (400) while other pressure ports (420) are positioned on a bottom region of process chip (400).

[0109] FIGS. 9-11 show first layer (500) in greater detail. First layer (500) is in the form of a rigid plate in this example. In some (but not necessarily all) versions, first layer (500) may be substantially translucent to visible light and / or ultraviolet light. By “substantially translucent” is meant that at least 90% (including in some instances 100%) of light is transmitted through the material compared to a translucent material. First layer (500) may comprise glass, plastic, silicone, and / or any other suitable material(s). In some versions, first layer (500) is formed as a lamination of two or morelayers of material, such that first layer (500) does not necessarily need to be formed as a single homogenous continuum of material.

[0110] As shown in FIGS. 9-11, first layer (500) includes an upper surface (502), a lower surface (504). a first plurality of openings (510) formed through both surfaces (502, 504), and a second plurality of openings (520) formed through both surfaces (502, 504). Openings (510) form part of fluid ports (410), such that each fluid port (410) has a respective opening (510), and such that openings (510) are configured to accommodate fluid lines (206). Openings (520) form part of pressure ports (420), such that each pressure port (420) has a respective opening (520), and such that openings (520) are configured to accommodate pressure lines (208). A plurality of process chamber portions (552) are formed as recesses in lower surface (504), collectively- providing a protruding region (550) in upper surface (502). Each process chamber portion (552) forms a "‘dry” region of a respective process chamber (450, 460, 470a, 470b) as described herein.

[0111] In the example shown, the plurality of openings (510) includes a pair of purification bead openings (510a, 510b) that are configured to allow communication of purification beads (e.g., together with a liquid solution in which the beads may be suspended) through first layer (500). First purification bead opening (510a) of the present example forms part of first purification bead port (510a), while second purification bead opening (510b) of the present example forms part of second purification bead port (510b), such that each purification bead port (410a, 410b) has a respective opening (510a, 510b).

[0112] As best seen in FIGS. 10-11, lower surface (504) of first layer (500) further defines a plurality of pneumatic channels (522), which are formed as recesses in lower surface (504). Each pneumatic channel (522) is in pneumatic communication with a corresponding opening (520). Each pneumatic channel (522) is further configured to define a space between first layer (500) and second layer (600), such that pressurized gas may be communicated along pneumatic channels (522).

[0113] Lower surface (504) of first layer (500) further defines a plurality of valve chamber portions (530) and a plurality of pump chamber portions (540). Valvechamber portions (530) and pump chamber portions (540) are formed as recesses in lower surface (504); and are in pneumatic communication with corresponding pneumatic channels (522). Pressurized gas may thus be communicated from pressure ports (420) to valve chamber portions (530) and pump chamber portions (540) via respective pneumatic channels (522).

[0114] Valve chamber portions (530) form “dry"’ regions of valve chambers as described herein; while pump chamber portions (540) form "dry" regions of pump chambers as described herein. In the present example, valve chamber portions (530) have 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, though other shapes may be used. By way of further example only, valve chambers associated with valve chamber portions (530) may be operated like chambers (320. 322) described above; while pump chambers associated with pump chamber portions (540) may be operated like chambers (322, 326) described above. While each chamber portion (530, 540) has its own associated pneumatic channel (522) and pressure port (420) in this example, each process chamber portion (552) also has its own associated pneumatic channel (522) and pressure port (420) in this example. Thus, each chamber portion (530, 540, 552) may be pneumatically pressurized independently of the other chamber portions (530, 540, 552).

[0115] Second layer (600) is interposed between first layer (500) and third layer (700). Second layer (600) of this example is in the form of first and second liquid- impermeable flexible membranes (600a, 600b) laminated to each other in a vertically stacked arrangement. As described below, second layer (600) may be used to drive fluids across process chip (400) via peristaltic pumping action. As also described below, second layer (600) may be used to provide valves at various locations along process chip (400). By way of example only, one or both flexible membranes (600a, 600b) of second layer (600) may comprise polydimethylsilicone (PDMS) elastomer film. Alternatively, any other suitable material(s) may be used to form second layer (600). While first flexible membrane (600a) is shown disposed below second flexible membrane (600b) such that first flexible membrane (600a) may be referred to as lower flexible membrane (600a) and second flexible membrane (600b) may be referred to asupper flexible membrane (600b), first flexible membrane (600a) may alternatively be disposed above second flexible membrane (600b).

[0116] Second layer (600) may be configured and operable like elastic layer (302) described above. Second layer (600) may thus pass through chambers collectively defined by first and third layers (500, 700) to separate such chambers into an upper chamber region (which receives pneumatic pressure) and a lower chamber region (which receives fluid). Such chambers include process chambers (450, 460, 470a, 470b), valve chambers as described herein, and pump chambers as described herein. Second layer (600) may further thus be pneumatically deformed to provide valving, peristaltic pumping, etc. within process chip (400).

[0117] As shown in FIGS. 12 and 13, second layer (600) includes a plurality of openings (602a, 602b) formed through flexible membranes (600a, 600b). Openings (602a, 602b) may be positioned at fluid ports (410) and / or pressure ports (420), under corresponding openings (510, 520), to allow fluid and / or pressurized gas to be communicated through second layer (600) at fluid ports (410) and / or pressure ports (420), respectively. For example, as noted above, each fluid port (410) is configured to receive a fluid line (206) from fluid interface assembly (109). The distal end of fluid line (206) may be configured to seal against the region of second layer (600) that is exposed by fluid port (410) and communicate fluid through the respective openings (602a, 602b) in second layer (600). In some versions, a spring or other resilient member provides a resilient bias to fluid line (206), urging the distal end of fluid line (206) against the region of second layer (600) that is exposed by fluid port (410) to thereby maintain the seal. Fluid from fluid line (206) may be further communicated through process chip (400) as described in greater detail below.

[0118] As shown in FIG. 12, second flexible membrane (600b) further includes a plurality of apertures (e.g., cutouts) (604) that are each configured to vertically align with a respective process chamber (450, 460, 470a, 470b), valve chamber, or pump chamber. For example, each aperture (604) may be disposed directly below the corresponding upper chamber region and / or directly above the corresponding lower chamber region. Each aperture (604) of second flexible membrane (600b) may directlyoverlie or directly underlie a corresponding portion of first flexible membrane (600a) that is configured to deflect within the respective process chamber (450, 460, 470a. 470b), valve chamber, or pump chamber. Apertures (604) may thereby accommodate deflection of first flexible membrane (600a) by preventing second flexible membrane (600b) of second layer (600) from forming wrinkles or otherwise interfering with the deflection of first flexible membrane (600a) of second layer (600) within process chambers (450, 460, 470a, 470b), valve chambers, or pump chambers.

[0119] By way of further example only, second layer (600) may be configured and operable in accordance with at least some of the teachings of US Pat. App. No. 63 / 453,206, entitled ‘'Microfluidic Apparatus with Elastic Layers and Contoured Surface,’" filed on March 20, 2023, the disclosure of which is incorporated by reference herein in its entirety.

[0120] In the present example, the region of each chamber in process chip (400) that is above second layer (600) constitutes a ‘"dry” chamber region since this region receives pressurized gas to pneumatically deflect second layer (600). Conversely, the region of each chamber in process chip (400) that is below second layer (600) constitutes a “wet” chamber region since this region receives fluid. However, these “wet” and “dry” roles may be reversed in some variations. In other words, some variations of process chip (400) may provide 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 upper chamber constitutes a “wet” chamber region.

[0121] FIGS. 14-18 show third layer (700) in greater detail. Third layer (700) is in the form of a rigid plate in this example. In some (but not necessarily all) versions, third layer (700) is substantially translucent to visible light and / or ultraviolet light. Third layer (700) may comprise glass, plastic, silicone, and / or any other suitable material(s). In some versions, third layer (700) is formed as a lamination of two or more layers of material, such that third layer (700) does not necessarily need to be formed as a single homogenous continuum of material.

[0122] As shown in FIGS. 12-14, third layer (700) includes an upper surface (702), a lower surface (704), and a plurality of openings (710) formed through both surfaces (702, 704). A plurality of process chamber portions (752) are formed as recesses in upper surface (702), collectively providing a protruding region (750) in lower surface (704). Each process chamber portion (752) forms a ‘"wet” region of a respective process chamber (450, 460, 470a, 470b) as described herein. Some of openings (710) form part of pressure ports (420), such that each pressure port (420) has a respective opening (710), and such that openings (710) are configured to allow communication of pressurized gas through third layer (700). As noted above, corresponding openings (602a, 602b) may be formed through second layer (600), such that at least some of openings (602a, 602b) in second layer (600) are between openings (520) of first layer (500) and at least some of openings (710) of third layer (700).

[0123] In the example shown, the plurality of openings (710) includes a plurality of purification bead openings (710a, 710b, 710c, 710c) that are configured to allow communication of purification beads (e.g., together with a liquid solution in which the beads may be suspended) through third layer (700). First purification bead opening (710a) of the present example forms part of first purification bead port (410a), while fourth purification bead opening (71 Od) of the present example forms part of second purification bead port (410b), such that each purification bead port (410a, 410b) has a respective opening (710a, 710d).

[0124] Each process chamber portion (752) has a sidewall (753) that extends downwardly from upper surface (702) tow ard a corresponding floor (755). As seen in FIGS. 14 and 16. upper surface (702) of third layer (700) further defines a plurality of fluid channels (712), which are formed as recesses in upper surface (702). Each fluid channel (712) is in fluid communication with a corresponding opening (710). Each fluid channel (712) is further configured to define a space betw een third layer (700) and second layer (600). such that fluid may be communicated along fluid channels (712).

[0125] In the example shown, the plurality of fluid channels (712) includes a plurality of purification bead channels (712a, 712b. 712c, 712d, 712e) that are configured to provide a path for communication of purification beads (e.g., togetherwith a liquid solution in which the beads may be suspended) to / from the “wet"’ region of first purification chamber (470a). In this regard, purification bead channels (712a. 712b, 712c, 712d, 712e) may each be sized and configured to accommodate flow of beads having a predetermined particle size (e.g., diameter) therethrough while mitigating against any risk of such beads becoming clogged therein or otherwise impeding fluid flow therethrough. To that end, purification bead channels (712a, 712b, 712c, 712d, 712e) may each have a cross dimension (e.g., width, depth, diameter, etc.) that is substantially greater than the corresponding cross dimension of some or all of the other fluid channels (712) formed in upper surface (702). In some cases, purification bead channels (712a, 712b, 712c, 712d, 712e) may each have a minimum cross dimension that is at least five times larger than the predetermined particle size of each bead. In scenarios where each bead has a predetermined particle size of about 50 pm, the minimum cross dimension of each purification bead channel (712a, 712b, 712c, 712d, 712e) may be at least about 250 pm. Each purification bead channel (712a, 712b, 712c. 712d, 712e) may have a width ranging from about 200 pm to about 800 pm, such as a width ranging from about 200 pm to about 750 pm, and may have a depth (e.g., relative to upper surface (702)) ranging from about 200 pm to about 750 pm; while some or all of the other fluid channels (712) formed in upper surface (702) may have a width ranging from about 100 pm to about 400 pm, and may have a depth (e.g., relative to upper surface (702)) ranging from about 100 pm to about 500 pm. For example, each purification bead channel (712a, 712b, 712c, 712d, 712e) may have a width of about 500 pm and a depth (e.g., relative to upper surface (702)) of about 250 pm; while some or all of the other fluid channels (712) formed in upper surface (702) may have a width of about 200 pm and a depth (e.g., relative to upper surface (702)) of about 100 pm. As another example, each purification bead channel (712a, 712b, 712c, 712d, 712e) may have a width of about 500 pm and a depth of about 500 pm. As yet another example, each purification bead channel (712a, 712b, 712c, 712d, 712e) may have a width of about 800 pm and a depth of about 400 pm. Alternatively, each purification bead channel (712a, 712b, 712c, 712d, 712e) may have any other suitable cross dimension(s) selected to accommodate beads of any predetermined particle size.

[0126] Upper surface (702) of third layer (700) further defines a plurality of valve chamber portions (730) and a plurality of pump chamber portions (740). Valve chamber portions (730) and pump chamber portions (740) are formed as recesses in upper surface (702); and are in fluid communication with corresponding fluid channels (712). Fluid may thus be communicated from fluid ports (410) to valve chamber portions (730) and pump chamber portions (740) via respective openings (710) and fluid channels (712).

[0127] Valve chamber portions (730) form “wet” regions of valve chambers as described herein (opposite to a corresponding valve chamber portion (530)); while pump chamber portions (740) form “wet” regions of pump chambers (opposite to a corresponding pump chamber portion (540)) as described herein. In the present example, valve chamber portions (730) have 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, though other shapes may be used. By way of further example only, valve chambers associated with valve chamber portions (730) may be operated like chambers (320, 322) described above; while pump chambers associated with pump chamber portions (740) may be operated like chambers (322. 326) described above. Each chamber portion (730, 740) has its own associated fluid channel (712) and opening (710) in this example. Thus, each chamber portion (730, 740) may receive fluid independently of the other chamber portions (730. 740).

[0128] Each process chamber portion (752) also has at least one respective fluid channel (712) leading thereto, such that fluid may be communicated to / from each process chamber (450, 460. 470a, 470b) via the fluid channel (712) leading to the process chamber portion (752) associated with the process chamber (450, 460, 470a, 470b). Also in this example, a pump chamber portion (740) and set of valve chamber portions (730) is interposed between an opening (710) and each fluid channel (712) leading to each respective process chamber portion (752). The pumps and valves defined by these portions (740, 730) may thus drive and regulate the communication of fluid to / from each process chamber (450, 460, 470a, 470b).

[0129] In the example shown, purification bead channels (712c, 712d) lead to the process chamber portion (752) associated with first purification chamber (470a). such that purification beads may be communicated to / from first purification chamber (470a) via 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 into first purification chamber (470a) for communicating purification beads to first purification chamber (470a), and purification bead channel (712d) may define a purification bead outlet channel (also referred to as a “particle outlet channef’) leading out of first purification chamber (470a) for communicating purification beads from first purification chamber (470a).

[0130] Also in this example, the plurality of valve chamber portions (730) includes a plurality of purification bead valve chamber portions (730a, 730b). and the plurality of pump chamber portions (740) includes a purification bead pump chamber portion (740a), that are each interposed between a respective opening (710) and a respective one of the purification bead channels (712c, 712d) leading to the process chamber portion (752) associated with first purification chamber (470a). The pump and valves defined by these portions (740a, 730a. 730b) may thus drive and regulate the communication of purification beads to / from first purification chamber (470a). In this regard, each purification bead valve chamber portion (730a, 730b) is disposed opposite to a corresponding purification bead valve chamber portion (530a, 530b) of the plurality of valve chamber portions (530). while purification bead pump chamber portion (740a) is disposed opposite to a purification bead pump chamber portion (540a) of the plurality of pump chamber portions (540).

[0131] The valves defined by purification bead valve chamber portions (530a, 530b, 730a, 730b) and the pump defined by purification bead pump chamber portions (540a, 740a) may each be sized and configured to accommodate flow of beads (e.g., together with a liquid solution in which the beads may be suspended) therethrough while mitigating against any risk of such beads becoming clogged therein or otherwise impeding fluid flow therethrough. For example, purification bead valve chamber portions (530a, 530b, 730a, 730b) may each have a cross dimension (e g., diameter) greater than that of some or all of the other valve chamber portions (530, 730). suchthat the valves defined by purification bead valve chamber portions (530a, 530b, 730a, 730b) may be configured to hold larger volumes of fluid than the valves defined by some or all of the other valve chamber portions (530, 730). In some cases, purification bead valve chamber portions (530a, 530b, 730a, 730b) may each have a diameter ranging from about 500 pm to about 6 mm, such as a diameter of about 2. 1 mm.

[0132] As shown, rather than having an oblong or stadium shape like that of some or all of the other pump chamber portions (540, 740), purification bead pump chamber portions (540a. 740a) of this example each have a circular shape, such that purification bead pump chamber portions (540a, 740a) are each shaped and / or sized similarly to valve chamber portions (530, 730). In addition, or alternatively, purification bead pump chamber portions (540a, 740a) may each have a depth (e.g., relative to the respective upper surface (502, 702)) greater than that of some or all of the other pump chamber portions (540, 740). In some cases, the relatively large depths of purification bead pump chamber portions (540a, 740a) may allow the pump defined by purification bead pump chamber portions (540a, 740a) to hold larger volumes of fluid than the pumps defined by some or all of the other pump chamber portions (540, 740). In some other cases, the relatively large depths of purification bead pump chamber portions (540a. 740a) in combination with the relatively small cross dimensions of purification bead pump chamber portions (540a, 740a) (e.g., due to purification bead pump chamber portions (540a. 740a) each having a circular shape rather than an oblong or stadium shape) may result in the pump defined by purification bead pump chamber portions (540a, 740a) being configured to hold substantially the same volume of fluid as the pumps defined by some or all of the other pump chamber portions (540, 740). For example, the pump defined by purification bead pump chamber portions (540a, 740a) and the pumps defined by some or all of the other pump chamber portions (540. 740) may each be configured to hold a volume of fluid ranging from about 50 nL to about 140 pL, such as about 3pL of fluid. In such cases, the relatively large depths of purification bead pump chamber portions (540a, 740a) may nevertheless assist with mitigating against any risk of beads becoming clogged in the pump defined by purification bead pump chamber portions (540a, 740a).

[0133] As also seen in FIGS. 14 and 16, third layer (700) includes a plurality of bridging channels (760). sidewall channels (754). and floor channels (756). Sidewall channels (754) may also be referred to as “sippers.” Each bridging channel (760) is formed as a recess in upper surface (702); each floor channel (756) is formed as a recess in a corresponding floor (755); and each sidewall channel (754) is formed as a recess in a corresponding sidewall (753) that extends upwardly from the corresponding floor channel (756) toward upper surface (702). The presence of sidewall channels (754) and / or floor channels (756) may provide a pathway for fluid to escape the respective process chamber (450, 460, 470a, 470b) when second layer (600) is deformed downwardly toward the respective floor (755), such that sidewall channels (754) and / or floor channels (756) may prevent pockets of fluid from being trapped between localized regions of deformed second layer (600) and the respective floor (755) and / or sidewall (753) (or otherwise reduce the risk of such pockets being formed).

[0134] As shown, at least some sidewall channels (754) extend from the corresponding floor channel (756) to a corresponding bridging channel (7 0), 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 communication of fluid from one process chamber (450, 460, 470a, 470b) to an adjacent process chamber (450, 460, 470a, 470b).

[0135] In the example shown, the plurality of bridging channels (760) includes a pair of purification bridging channels (760a, 760b) extending between the process chamber portions (752) associated with purification chambers (470a, 470b). In some versions, purification bridging channels (760a, 760b) are configured to provide a path for communication of purification beads (e g., together with a liquid solution in which the beads may be suspended) between purification chambers (470a, 470b). In this regard, purification bridging channels (760a, 760b) may each be sized and configured to accommodate flow of beads therethrough while mitigating against any risk of such beads becoming clogged therein or otherwise impeding fluid flow therethrough. For example, each purification bridging channel (760a, 760b) may have a width ranging from about 250 pm to about 750 pm, such as a width of about 500 pm, and a depth (e.g., relative to upper surface (702)) ranging from about 250 pm to about 750 pm, suchas a depth of about 500 pm. Alternatively, each purification bridging channel (760a, 760b) may have any other suitable cross dimension(s) selected to accommodate beads of any predetermined particle size.

[0136] As shown in FIG. 15. third layer (700) further includes a plurality of fluid channels (714) on lower surface (704). Fluid channels (714) are formed as recesses on lower surface (704). Each fluid channel (714) is further configured to define a space between third layer (700) and fourth layer (800), such that fluid may be communicated along fluid channels (714). An opening (710) is positioned at each end of each fluid channel (714). Openings (710) and fluid channels (714) thus cooperate to allow' fluid to be communicated from one region at upper surface (702) of third layer (700) to another region at upper surface (702) of third layer (700) via lower surface (704) of third layer (700), which may facilitate routing of fluid more easily than could otherwise be achieved if the fluid were only routed via fluid channels (712) in upper surface (702). In other words, the inclusion of fluid channels (714) on low er surface (704) may allow' fluid to be routed along a path that would otherwise intersect the fluid path provided via a fluid channel (712) if such path were moved from lower surface (704) to upper surface (702).

[0137] In the example shown, the plurality of fluid channels (714) includes a plurality of purification bead channels (714a, 714b) that are sized and configured similarly to purification bead channels (712a, 712b, 712c, 712d, 712e), to accommodate flow' of beads having a predetermined particle size (e.g., diameter) therethrough while mitigating against any risk of such beads becoming clogged therein or otherwise impeding fluid flow therethrough. First purification bead channel (714a) of the present example extends from first purification bead opening (710a) to second purification bead opening (710b) (which communicates with first purification bead channel (712a) on upper surface (702)), while second purification bead channel (714b) of the present example extends from fourth purification bead opening (71 Od) to third purification bead opening (710c) (which communicates with fifth purification bead channel (712e) on upper surface (702)).

[0138] Referring now to FIGS. 17 and 18, with continuing reference to FIGS. 14 and 16. third layer (700) of the present example also includes a plurality of purification bead filters (770a, 770b, 770c) on upper surface (702). Purification bead filters (770a, 770b, 770c) may also be referred to as “fritting structures.” Purification bead filters (770a, 770b, 770c) are each interposed between a respective one of the process chamber portions (752) associated with purification chambers (470a. 470b) and one or more corresponding fluid channels (712) (e.g., one or more corresponding fluid outlet channels). More particularly, a first purification bead filter (770a) is interposed between the process chamber portion (752) associated with first purification chamber (470a) and a plurality of corresponding fluid channels (712) that define fluid outlet channels for communicating a fluid output from first purification chamber (470a); a second purification bead filter (770b) is interposed between the process chamber portion (752) associated with second purification chamber (470b) and a corresponding fluid channel (712) that defines a waste outlet channel for communicating a fluid waste from second purification chamber (470b); and a third purification bead filter (770c) is interposed between the process chamber portion (752) associated with second purification chamber (470b) and a corresponding fluid channel (712) that defines a UV sampling outlet channel for communicating a fluid sample from second purification chamber (470b). Purification bead filters (770a, 770b, 770c) are each configured to provide a path for communication of fluid from the respective purification chamber (470a, 470b) to the one or more corresponding fluid channels (712), while inhibiting the passage of beads from the respective purification chamber (470a, 470b) to the one or more corresponding fluid channels (712) to thereby retain such beads within purification chambers (470a. 470b).

[0139] In this regard, purification bead filters (770a, 770b, 770c) each include an array of filter channels (772) formed as recesses in upper surface (702), a filter entrance (774) formed as a recess in a corresponding sidewall (753) and extending between the inner ends of the respective filter channels (772), and a filter exit (776) formed as a recess in 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 entrance (774) and filter exit (776). In the example shown, filter entrance (774)of each purification bead filter (770a, 770b. 770c) extends along a peripheral edge of the respective process chamber portion (752); filter channels (772) of each purification bead filter (770a, 770b, 770c) extend substantially parallel to each other and substantially perpendicular to the corresponding filter entrance (774); and filter exit (776) of each purification bead filter (770a, 770b, 770c) extends substantially perpendicular to the corresponding filter channels (772) and substantially parallel to the corresponding filter entrance (774). Each filter channel (772) is further configured to define a space between third layer (700) and second layer (600), and each filter exit (776) is likewise configured to define a space between third layer (700) and second layer (600), such that fluid may be communicated along filter channels (772) and filter exits (776) to the corresponding one or more fluid channels (712).

[0140] Filter channels (772) may each be sized and configured to accommodate flow of fluid therethrough while preventing the flow of beads having a predetermined particle size (e.g., diameter) therethrough. In other words, filter channels (772) of each purification bead filter (770a, 770b, 770c) may be configured to cooperate with each other to function as a screen or “frit’' to filter out such beads from fluid flowing therethrough. To that end. filter channels (772) may each have a cross dimension (e.g.. width, depth, diameter, etc.) that is substantially less than the corresponding cross dimension of some or all of the fluid channels (712) formed in upper surface (702). For example, filter channels (772) may each be relatively shallow, at least by comparison to some or all of the fluid channels (712) formed in upper surface (702). In some cases, filter channels (772) may each have at least one cross dimension that is substantially less than the predetermined particle size of each bead. In scenarios where each bead has a predetermined particle size of about 50 pm, at least one cross dimension of each filter channel (772) may be substantially less than about 50 pm. For example, each filter channel (772) may have a depth (e.g., relative to upper surface (702)) ranging from about 10 pm to about 30 pm, such as a depth of about 25 pm. As another example, each filter channel (772) may have a depth ranging from about 10 pm to about 23 pm. Since one cross dimension of each filter channel (772), such as its depth, may be sufficiently small to prevent the flow of beads therethrough, the other cross dimension of each filter channel (772), such as its width, may in some cases be substantially equalto or even greater than the predetermined particle size of each bead. In scenarios where each bead has a predetermined particle size of about 50 pm, the other cross dimension of each filter channel (772) may be substantially equal to or greater than about 50 pm. For example, each filter channel (772) may have a width ranging from about 100 pm to about 400 pm, such as a width of about 200 pm. Alternatively, each filter channel (772) may have any other suitable cross dimension(s). such as any other suitable depth(s), selected to filter out beads of any predetermined particle size.

[0141] The presence of a filter entrance (774) for each purification bead filter (770a, 770b, 770c) may assist with mitigating against any risk of beads blocking fluid from reaching the respective filter channels (772) by providing space for beads to collect without covering the respective filter channels (772). In addition, or alternatively, the presence of multiple filter channels (772) for each purification bead filter (770a, 770b, 770c) may reduce the impact on fluid flow of any blockages that might occur over a particular filter channel (772) by providing one or more alternative paths for fluid to reach the respective filter exit (776) (e.g., via one or more of the other, unblocked filter channels (772)). It will be appreciated that purification bead filters (770a, 770b, 770c) may each include any suitable number(s) of filter channels (772).

[0142] As shown, at least some sidewall channels (754) extend from the floor channel (756) of a process chamber portion (752) associated with one of purification chambers (470a, 470b) to a corresponding purification bead filter (770a, 770b, 770c). Such sidewall channels (754) may be configured to provide a path for communication of purification beads (e.g.. together with a liquid solution in which the beads may be suspended) from the respective purification chamber (470a, 470b) to the corresponding purification bead filter (770a, 770b, 770c). In this regard, such sidewall channels (754) may each be sized and configured to accommodate flow of beads therethrough while mitigating against any risk of such beads becoming clogged therein or otherwise impeding fluid flow therethrough. For example, each such sidewall channel (754) may have a width ranging from about 200 pm to about 800 pm, such as a width of about 500 pm, and a depth (e.g., relative to the corresponding sidewall (753)) ranging from about 100 pm to about 300 pm, such as a depth of about 100 pm. Alternatively, each sidewall channel (754) may have any other suitable cross dimension(s).

[0143] In the example shown, first purification bead filter (770a) is equipped with a plurality of (e.g., three) such sidewall channels (754). while 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 first purification bead filter (770a) may assist with mitigating against any risk of beads becoming clogged therein by providing multiple paths for such beads to first purification bead filter (770a). In addition, or alternatively, the presence of multiple sidewall channels (754) leading to first purification bead filter (770a) may reduce the impact on fluid flow of any clogs that might occur within a particular sidewall channel (754) by providing one or more alternative paths for fluid to reach first purification bead filter (770a) (e.g., via one or more of the other, unclogged sidewall channels (754)). It will be appreciated that purification bead filters (770a, 770b, 770c) may each be equipped with any other suitable number(s) of sidewall channels (754).

[0144] In the example shown, another sidewall channel (754) extends from the floor channel (756) of process chamber portion (752) associated with first purification chamber (470a) to purification bead outlet channel (712d). This sidewall channel (754) may be configured to provide a path for communication of fluid from first purification chamber (470a) to purification bead outlet channel (712d).

[0145] An example of a flow path of particles through a portion of process chip (400) is shown in FIG. 19. For example, such particles may include purification beads such as POROS Oligo (dT)25 Affinity' Resin by Thermo Fisher Scientific of Waltham, Massachusetts. The purification beads may each have a predetermined particle size (e.g.. diameter), such as ranging from about 10 pm to about 200 pm. such as about 50 pm, and / or may be suspended in a liquid solution (e g., 0.1 M sodium chloride) to define a slurry'. It will be appreciated that any other suitable purification beads or other ty pes of functionalized particles, such as urea granules, may follow' the flow path shown in FIG. 19.

[0146] As indicated by arrows (Al), purification beads may initially be communicated from first purification bead port (410a) (e.g., in cooperation with a first fluid line (206)) through first purification bead opening (710a), along first purificationbead channel (714a) on lower surface (704), through second purification bead opening (710b). along first purification bead channel (712a) on upper surface (702), through purification bead pump chamber portion (740a), along second purification bead channel (712b) on upper surface (702), through first purification bead valve chamber portion (730a), along third purification bead channel (712c) on upper surface (702), and into the “wet” region of first purification chamber (470a). Purification beads may treat fluid that is simultaneously delivered, that has been previously delivered, or that is subsequently delivered to the “wet” region of first purification chamber (470a).

[0147] As indicated by arrows (A2), purification beads may then be communicated from the “wet” region of first purification chamber (470a) to one or both purification bridging channels (760a. 760b) (e.g., via the respective sidewall channels (754)), and into the “wet” region of second purification chamber (470b). Purification beads may treat fluid that is simultaneously delivered, that has been previously delivered, or that is subsequently delivered to the “wet” region of second purification chamber (470b). As also indicated by arrows (A2), purification beads may also be communicated back- and-forth between the “wet" regions of first and second purification chambers (470a, 470b). In some instances, purification beads may be communicated back-and-forth between the “wet” regions of first and second purification chambers (470a, 470b) together with the fluid being treated.

[0148] As indicated by arrows (A3), purification beads may then be communicated from the “wet” region of first purification chamber (470a) to fourth purification bead channel (712d) on upper surface (702) (e.g., via the respective sidewall channel (754)), through second purification bead valve chamber portion (730b). along fifth purification bead channel (712e) on upper surface (702), through third purification bead opening (710c), along second purification bead channel (714b) on lower surface (704), through fourth purification bead opening (710d), and into second purification bead port (410b). Purification beads may be removed from process chip (400) via second purification bead port (410b) (e.g., in cooperation with a second fluid line (206)), and / or may be returned to first purification bead port (410a) for communication back to the “wet” region of first purification chamber (470a). It will be appreciated that second layer (600) may be pneumatically deformed in a manner similar to that discussed above toprovide appropriate valving and / or peristaltic pumping for communicating purification beads along each portion of the flow path indicated by arrows (Al, A2. A3).

[0149] Due to the presence of purification bead filters (770a. 770b, 770c) at each fluid outlet of purification chambers (470a, 470b), other than purification bridging channels (760a, 760b) and purification bead outlet channel (712d), purification beads may be substantially constrained to the flow path indicated by arrows (Al, A2, A3). Thus, any purification beads that might be suspended within fluid that is communicated from first purification chamber (470a) through first purification bead filter (770a) may be filtered out of such fluid and left behind in first purification chamber (470a); such purification beads may be subsequently removed from first purification chamber (470a) via purification bead outlet channel (712d). Similarly, any purification beads that might be suspended within fluid that is communicated from second purification chamber (470b) through second or third purification bead filters (770b, 770c) may be filtered out of such fluid and left behind in second purification chamber (470b); such purification beads may be subsequently communicated from second purification chamber (470b) to first purification chamber (470a) via purification bridging channels (760a, 760b), and then removed from first purification chamber (470a) via purification bead outlet channel (712d). In this manner, purification beads may be removed from any purified fluid exiting purification chambers (470a, 470b), and thus may be prevented from interfering with fluid flow and / or processing downstream of purification bead filters (770a, 770b, 770c).

[0150] While purification beads are shown only being routed to / from purification chambers (470a. 470b) for treating fluid therein in the present example, it will be appreciated that purification beads or other types of functionalized particles may additionally or alternatively be routed to / from any one or more of the other process chambers (450, 460) for treating fluid therein, as may be desired. For example, any one or more of process chambers (450, 460) may be equipped with corresponding bead channels similar to purification bead channels (712a, 712b, 712c, 712d, 712e) that are configured to provide a path for communication of corresponding beads (e.g., together with a liquid solution in which the beads may be suspended) to / from the “wet” region of the respective process chamber (450, 460); and with corresponding valves and / orpumps as described herein. In some cases, the any one or more of process chambers (450. 460) may also be equipped with corresponding bead filters similar to purification bead filters (770a, 770b, 770c) that are configured to provide a path for communication of fluid from the respective process chamber (450, 460) to one or more corresponding fluid channels (712), while inhibiting the passage of beads from the respective process chamber (450, 460) to the one or more corresponding fluid channels (712) to thereby retain such beads within the respective process chamber (450, 460).

[0151] While purification bead filters (770a, 770b, 770c) of the present example are each interposed between a respective one of the process chamber portions (752) associated with purification chambers (470a, 470b) and one or more corresponding fluid outlet channels (712) such that purification bead filters (770a, 770b, 770d) are each at or near a corresponding fluid outlet of the respective purification chamber (470a, 470b), it will be appreciated that one or more purification bead filters (770a, 770b, 770c) may additionally or alternatively be at or near a corresponding fluid inlet of the respective purification chamber (470a, 470b) or other process chamber (450, 460). For example, one or more purification bead filters (770a, 770b, 770c) may be interposed between a respective one of the process chamber portions (752) associated with process chambers (450, 460, 470a, 470b) and one or more corresponding fluid inlet channels (712). Such purification bead filter(s) (770a, 770b, 770c) may be configured to provide a path for communication of fluid from the one or more corresponding fluid channels (712) to the respective process chamber (450. 460, 470a, 470b), while inhibiting the passage of beads from the respective process chamber (450, 460, 470a, 470b) to the one or more corresponding fluid channels (712) to thereby retain such beads within the respective process chamber (450, 460, 470a, 470b). The presence of such purification bead filter(s) (770a, 770b, 770c) at or near a fluid inlet may assist with mitigating against any nsk of beads blocking fluid from reaching the respective one of the process chamber portions (752) associated with the respective process chamber (450, 460, 470a, 470b).

[0152] Referring again to FIG. 8, fourth layer (800) may include an elastomeric membrane, such as silicone and / or any other suitable material(s). As shown, fourth layer (800) of this example includes a central opening (802) that is sized and configuredto receive protruding region (750) of third layer (700). In the present example, fourth layer (800) provides a gasket forming a seal between adjacent regions of third layer (700) and fifth layer (900). In this regard, fifth layer (900) is in the form of a rigid plate in this example. As shown, fifth layer (900) of this example includes a central opening (902) that is sized and configured to receive protruding region (750) of third layer (700). In the example shown, sixth layer (1000) is sized and configured to directly underlie protruding region (750) of third layer (700), and seventh layer (1100) is sized and configured to directly underlie sixth layer (1000). In this regard, sixth layer (1000) is in the form of a gap thermal pad in this example, and seventh layer (1100) is in the form of a graphite thermal pad in this example.

[0153] It will be appreciated that any suitable structures or techniques may be used to secure layers (500, 600, 700, 800. 900, 1000, 1100) together.

[0154] While purification bead filters (770a, 770b, 770c) of the present example are each formed on upper surface (702) of third layer (700), it may be desirable to at least partially incorporate one or more purification bead filters (770a, 770b, 770c) into one or more other suitable layers of process chip (400), such as second layer (600). For example, doing so may reduce the cost and / or complexity of manufacturing third layer (700), such as in instances where the predetermined particle size of the purification beads is smaller than about 50 pm, which might otherwise entail the machining of very small (e.g., about 25 pm deep or about 10 pm deep) filter channels (772) into upper surface (702) of third layer (700).

[0155] FIG. 20 show s a portion of another example of a process chip (1400) that may provide at least some, if not all, of the features and functionalities described above. Process chip (1400) is similar to process chip (400) described above, except as otherwise described below. In this regard, process chip (1400) of this example includes a first layer (1500), a second layer (1600), and a third layer (1700), which may be similar to first layer (500), second layer (600), and third layer (700) described above, respectively, except as otherwise described below. While not shown, process chip (1400) may also include any one or more of a fourth layer similar to fourth layer (800); a fifth layer similar to fifth layer (900); a sixth layer similar to sixth layer (1000); aseventh layer similar to seventh layer (1100); and / or any other suitable features shown and / or described herein in connection with process chip (400).

[0156] As shown, first layer (1500) includes a lower surface (1504) similar to lower surface (504). While not shown, first layer (1500) may also include any one or more of an upper surface similar to upper surface (502); openings similar to openings (510, 520); pneumatic channels similar to pneumatic channels (522); valve chamber portions similar to valve chamber portions (530); pump chamber portions similar to pump chamber portions (540); a protruding region similar to protruding region (550); process chamber portions similar to process chamber portions (552); and / or any other suitable features shown and / or described herein in connection with first layer (500).

[0157] Second layer (1600) includes a lower flexible membrane (1600a) and an upper flexible membrane (1600b) similar to flexible membranes (600a, 600b), respectively. While not shown, second layer (1600) may include any one or more of openings similar to openings (602a, 602b); apertures similar to apertures (604); and / or any other suitable features shown and / or described herein in connection with second layer (600).

[0158] Third layer (1700) includes an upper surface (1702) similar to upper surface (702); a lower surface (1704) similar to lower surface (704); a plurality' of fluid channels (1712) (one shown) similar to fluid channels (712), that are formed as recesses in upper surface (1702); and a plurality of process chamber portions (1752) (one shown) similar to process chamber portions (752), that are formed as recesses in upper surface (1702). Each process chamber portion (1752) has a si de wall (1753) that extends downwardly from upper surface (1702) toward a corresponding floor (1755). In the example shown, a filter entrance (1774) is formed as a recess in a corresponding sidewall (1753). While not shown, third layer (1700) may include any one or more of openings similar to openings (710); fluid channels similar to fluid channels (714); valve chamber portions similar to valve chamber portions (730); pump chamber portions similar to pump chamber portions (740); a protruding region similar to protruding region (750); sidewall channels similar to sidewall channels (754); floor channels similar to floor channels(756); bridging channels similar to bridging channels (760); and / or any other suitable features shown and / or described herein in connection with third layer (700).

[0159] Second layer (1600) of the present example also includes a plurality of purification bead filters (1670) (one shown). Purification bead filters (1670) may also be referred to as “fritting structures." Purification bead filters (1670) are each interposed between a respective one of the process chamber portions (1752) and one or more corresponding fluid channels (1712) (e.g., one or more corresponding fluid inlet or outlet channels). Purification bead filters (1670) are each configured to provide a path for communication of fluid from the respective process chamber portion (1752) to the one or more corresponding fluid channels (1712), while inhibiting the passage of beads from the respective process chamber portion (1752) to the one or more corresponding fluid channels (1712) to thereby retain such beads within the process chamber portions (1752).

[0160] In this regard, purification bead filters (1670) each include at least one filter channel (1672) formed as an aperture (e.g., cutout) through upper flexible membrane (1600b), a filter entrance (1674) formed as a first slot through lower flexible membrane (1600a) and fluidically coupling an inner end of the at least one filter channel (1672) to filter entrance (1774), and a filter exit (1676) formed as a second slot through lower flexible membrane (1600a) and fluidically coupling an outer end of the at least one filter channel (1672) to the one or more corresponding fluid channels (1712), such that each filter channel (1672) extends along an upper surface of lower flexible membrane (1600a) between the respective filter entrance (1674) and filter exit (1676). Each filter channel (1672) is further configured to define a space between second layer (1600) (e.g., lower flexible membrane (1600a) thereol) and first layer (1500), such that fluid may be communicated along filter channels (1672) and filter exits (1676) to the corresponding one or more fluid channels (1712).

[0161] Filter channels (1672) may each be sized and configured to accommodate flow of fluid therethrough while preventing the flow of beads having a predetermined particle size (e.g., diameter) therethrough. In other words, filter channels (1672) of each purification bead filter (1670) may be configured to cooperate with each other tofunction as a “frit” to filter out such beads from fluid flowing therethrough. To that end, filter channels (1672) may each have a cross dimension (e.g., width, depth, diameter, etc.) that is substantially less than the corresponding cross dimension of some or all of the fluid channels (1712) formed in upper surface (1702). For example, filter channels (1672) may each be relatively shallow, at least by comparison to some or all of the fluid channels (1712) formed in upper surface (1702). In some cases, filter channels (1672) may each have at least one cross dimension that is substantially less than the predetermined particle size of each bead. In scenarios where each bead has a predetermined particle size of about 50 pm, at least one cross dimension of each filter channel (1672) may be substantially less than about 50 pm. For example, each filter channel (1672) may have a depth (e.g., relative to lower surface (1504)) ranging from about 10 pm to about 30 pm, such as a depth of about 1 pm or a depth of about 25 pm. As shown, the depth of each filter channel (1672) may be substantially equal to a thickness of upper flexible membrane (1600b). Thus, in cases where upper flexible membrane (1600b) has athickness of about 10 pm, each filter channel (1672) may have a depth of about 10 pm. Since one cross dimension of each filter channel (1672), such as its depth, may be sufficiently small to prevent the flow of beads therethrough, the other cross dimension of each filter channel (1672), such as 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 dimension(s), such as any other suitable depth(s), selected to filter out beads of any predetermined particle size.

[0162] The presence of a filter entrance (1774) for each purification bead filter (1670) may assist with mitigating against any risk of beads blocking fluid from reaching the respective filter channel (1672) by providing space for beads to collect without covering the respective filter channel (1672). In addition, or alternatively, the filter channel (1672) for each purification bead filter (1670) may have a sufficiently large width to reduce the impact on fluid flow of any blockages that might occur over the filter channel (1672) by providing one or more alternative paths for fluid to reach the respective filter exit (1676) (e.g., by circumnavigating the blockage within the filterchannel (1672)). It will be appreciated that purification bead filters (1670) may each include any suitable number(s) of filter channels (1672).

[0163] Particles may flow through process chip (1400) along a flow path similar to that shown in FIG. 19. It will be appreciated that second layer (1600) may be pneumatically deformed in a manner similar to that discussed above to provide appropriate valving and / or peristaltic pumping for communicating purification beads along each portion of the flow path.

[0164] Due to the presence of purification bead filter (1670) at a fluid outlet of the illustrated process chamber portion (1752), any purification beads that might be suspended within fluid that is communicated from the illustrated process chamber portion (1752) through the illustrated purification bead filter (1670) may be filtered out of such fluid and left behind in the illustrated process chamber portion (1752). In this manner, purification beads may be removed from any purified fluid exiting the illustrated process chamber portion (1752), and thus may be prevented from interfering with fluid flow and / or processing downstream of the illustrated purification bead filter (1670).

[0165] FIG. 21 shows a portion of another example of a process chip (2400) that may provide at least some, if not all, of the features and functionalities described above. Process chip (2400) is similar to process chip (400) described above, except as otherwise described below. In this regard, process chip (2400) of this example includes a first layer (not shown) similar to first layer (500), a second layer (not shown) similar to second layer (600), and a third layer (2700), which may be similar to third layer (700) described above, except as otherwise described below. While not shown, process chip (2400) may also include any one or more of a fourth layer similar to fourth layer (800); a fifth layer similar to fifth layer (900); a sixth layer similar to sixth layer (1000); a seventh layer similar to seventh layer (1100); and / or any other suitable features shown and / or described herein in connection with process chip (400).

[0166] As shown, third layer (2700) includes an upper surface (2702) similar to upper surface (702); a plurality of fluid channels (2712) similar to fluid channels (712), that are formed as recesses in upper surface (2702); a plurality of valve chamberportions (2730) similar to valve chamber portions (730), that are formed as recesses in upper surface (2702); a plurality of process chamber portions (2752) similar to process chamber portions (752), that are formed as recesses in upper surface (2702). Each process chamber portion (2752) has a sidewall (2753) that extends downwardly from upper surface (2702) toward a corresponding floor (2755). As also seen in FIG. 21, third layer (2700) includes sidewall channels (2754) similar to sidewall channels (754), and floor channels (2756) similar to floor channels (756). While not shown, third layer (2700) may include any one or more of openings similar to openings (710); fluid channels similar to fluid channels (714); pump chamber portions similar to pump chamber portions (740); a protruding region similar to protruding region (750); bridging channels similar to bridging channels (760); and / or any other suitable features shown and / or described herein in connection with third layer (700).

[0167] In the example show n, the plurality of fluid channels (2712) includes at least one fluid inlet channel (2712a) leading into a corresponding process chamber portion (2752) for communicating fluid to the corresponding process chamber portion (2752), at least one fluid outlet channel (2712b) for communicating fluid from the corresponding process chamber portion (2752), and at least one backflush channel (2712c), the purpose of which will be described below. Also in this example, the plurality of valve chamber portions (2730) includes at least one fluid inlet valve chamber portion (2730a) upstream of the at least one fluid inlet channel (2712a), at least one fluid outlet valve chamber portion (2730b) downstream of the at least one fluid outlet channel (2712b), and at least one backflush valve chamber portion (2730c) positioned along the at least one backflush channel (2712c).

[0168] Third layer (2700) of the present example also includes a plurality’ of purification bead filters (2770a, 2770b) on upper surface (2702). Purification bead filters (2770a, 2770b) may also be referred to as “fritting structures.” Purification bead filters (2770a, 2770b) are each interposed between a respective one of the process chamber portions (2752) and one or more corresponding fluid outlet channels (2712b). Purification bead filters (2770a, 2770b) are each configured to provide a path for communication of fluid from the respective process chamber portion (2752) to the one or more corresponding fluid outlet channels (2712b), while inhibiting the passage ofbeads from the respective process chamber portion (2752) to the one or more corresponding fluid outlet channels (2712b) to thereby retain such beads within the process chamber portions (2752).

[0169] In this regard, purification bead filters (2770a, 2770b) each include an array of filter channels (2772) formed as recesses in upper surface (2702), a filter entrance (2774) formed as a recess in a corresponding sidewall (2753) and extending between the inner ends of the respective filter channels (2772), and a filter exit (2776) formed as a recess in 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 entrance (2774) and filter exit (2776). Each filter channel (2772) is further configured to define a space between third layer (2700) and second layer (600), and each filter exit (2776) is likewise configured to define a space between third layer (2700) and second layer (600), such that fluid may be communicated along filter channels (2772) and filter exits (2776) to the corresponding one or more fluid outlet channels (2712b).

[0170] Filter channels (2772) may each be sized and configured to accommodate flow of fluid therethrough while inhibiting the flow of beads having a predetermined particle size (e.g., diameter) therethrough. In other words, filter channels (2772) of each purification bead filter (2770a, 2770b) may be configured to cooperate with each other and / or with the beads themselves to function as a screen or “frit” to filter out such beads from fluid flowing therethrough. To that end, filter channels (2772) may each have a cross dimension (e.g.. width, depth, diameter, etc.) that is substantially less than the corresponding cross dimension of some or all of the fluid channels (2712) formed in upper surface (2702). While each individual filter channel (2772) may have a cross- sectional area that is smaller than that some or all of the fluid channels (2712), the plurality of filter channels (2772) of a particular purification bead filter (2770a, 2770b) may have a collective cross-sectional area that is substantially equal to or greater than the cross-sectional area of some or all of the fluid channels (2712). For example, filter channels (2772) may each be relatively shallow, at least by comparison to some or all of the fluid channels (2712) formed in upper surface (2702). In some cases, filter channels (2772) may each have at least one cross dimension that is substantially greaterthan the predetermined particle size of each bead. For example, fdter channels (2772) may each have at least one cross dimension that is greater than or equal to about twice the predetermined particle size of each bead and / or that is less than about five times the predetermined particle size of each bead. In scenarios where each bead has a predetermined particle size of about 50 pm, at least one cross dimension of each filter channel (2772) may be substantially greater than about 50 pm. For example, each filter channel (2772) may have a depth (e.g., relative to upper surface (2702)) of greater than or equal to about 100 pm. Even with one cross dimension of each filter channel (2772), such as its depth, being substantially greater than the predetermined particle size of each bead, the other cross dimension of each filter channel (2772), such as its width, may also be substantially greater than the predetermined particle size of each bead. In scenarios where each bead has a predetermined particle size of about 50 pm, the other cross dimension of each filter channel (2772) may be substantially equal to or greater than about 50 pm. For example, each filter channel (2772) may have a width ranging from about 100 pm to about 400 pm, such as a width of about 200 pm.

[0171] Thus, filter channels (2772) may be substantially larger than the beads. Nevertheless, filter channels (2772) may be configured to inhibit the flow of beads therethrough via jamming of at least some beads therein. This jamming may assist with inhibiting the flow of beads while still allowing the flow of fluid through filter channels (2772), such as in cases where there is some variation in sizes among 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 50 pm ± 10 pm. In such scenarios, larger beads may cooperate with smaller beads to jam within filter channels (2772) while defining openings between the beads that are sufficiently large to allow the flow of fluid therethrough and that are sufficiently small to prevent the flow of other beads therethrough. Thus, the beads may be retained within the process chamber portions (2752) despite each individual bead being substantially smaller than filter channels (2772). In some instances, a substantially minimal amount of beads may be capable of escaping a filter channel (2772). For example a substantially minimal amount of beads may be capable of escaping a filter channel (2772) before a jam is formed therein. In addition, or alternatively, a substantiallyminimal amount of beads may be capable of escaping a filter channel (2772) after a jam is formed therein by escaping through one or more openings defined by the jam. But due to the jamming of beads within filter channels (2772), only a single-digit percentage (e.g., only about 1%) of the beads contained within a process chamber portions (2752) may be capable of escaping out of the process chamber portions (2752) through purification bead filters (2770a, 2770b). It will also be appreciated that the increased size of filter channels (2772) relative to beads may assist with mitigating against any risk of beads blocking fluid from flowing through each filter channel (2772).

[0172] In the example shown, the fluid pathways that are downstream of filter channels (2772) are substantially free of potential blockage areas to avoid jamming of any beads that might escape from the corresponding filter channels (2772). For example, process chip (2400) of the present example lacks any vacuum caps between each purification bead filter (2770a, 2770b) and the corresponding fluid outlet valve chamber portion (2730b). In addition, or alternatively, each filter exit (2776) and / or each fluid outlet channel (2712b) may be sized to avoid jamming of any beads that might escape thereto from the corresponding filter channels (2772). For example, each cross dimension (e.g., width and depth) of each filter exit (2776) and / or of each fluid outlet channel (2712b) may be greater than about five times the predetermined particle size of each bead. In scenarios where each bead has a predetermined particle size of about 50 pm, each filter exit (2776) and / or each fluid outlet channel (2712b) may have a width of about 500 pm and / or a depth of about 250 pm.

[0173] Similarly, each fluid inlet channel (2712a) may be sized to avoid jamming of any beads that might flow upstream thereto from the corresponding process chamber portion (2752). For example, each cross dimension (e.g., width and depth) of each fluid inlet channel (2712a) may be greater than about five times the predetermined particle size of each bead. In scenarios where each bead has a predetermined particle size of about 50 pm. each fluid inlet channel (2712a) may have a width of about 500 pm and / or a depth of about 250 pm. Thus, fluid inlet channels (2712a) may have an increased size relative to at least some of the other fluid channels (2712) on third layer (2700), such as the fluid channel (2712) that is upstream of the corresponding fluid inlet valve chamber portion (2730a).

[0174] In the example shown, each backflush channel (2712c) is downstream of a corresponding filter exit (2776) such that each backflush channel (2712c) is configured to facilitate backflushing of the corresponding filter channels (2772), such as in cases where one or more of the corresponding filter channels (2772) has been substantially clogged by beads. For example, with the valve associated with backflush valve chamber portion (2730c) in an open state and with the valve associated with fluid outlet valve chamber portion (2730b) in a closed state, a backflush fluid (e.g., water) may flow in an upstream direction from backflush channel (2712c) into the corresponding filter exit (2776) and through the corresponding filter channels (2772) to push any beads that might be clogged within the corresponding filter channels (2772) back into the corresponding process chamber portion (2752). Such backflushing of filter channels (2772) may be performed in accordance with a predetermined routine based on certain criteria. For example, such backflushing may be performed in response to a duration of fluid flowing through the corresponding filter (2770a, 2770b) reaching a predetermined threshold duration; and / or in response to a cumulative volume of fluid flowed through the corresponding filter (2770a, 2770b) reaching a predetermined threshold volume; and / or in response to a number of cycles of fluid flowing through the corresponding filter (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 a flow blockage within the corresponding filter (2770a, 2770b).

[0175] In addition, or alternatively, each backflush channel (2712c) may be configured to facilitate flushing of the entire fluid path from the corresponding process chamber portion (2752) to the corresponding destination vial or other fluid storage container (e.g., in reagent storage frame (107)). For example, with the valve associated with backflush valve chamber portion (2730c) in an open state, and with the valve associated with fluid outlet valve chamber portion (2730b) in an open state, and with the process chamber associated with process chamber portion (2752) in a closed state, the backflush fluid (e.g.. water) may flow in a downstream direction from backflush channel (2712c) into the corresponding fluid outlet channel (2712b) and through the corresponding fluid outlet valve chamber portion (2730b) all the way to thecorresponding destination vial / container to push any beads or fluid disposed along the fluid path to the corresponding destination vial / container.

[0176] VI. Examples of Combinations

[0177] The following examples relate to various non-exhaustive ways in which the teachings herein may be combined or applied. It should be understood that the following examples are not intended to restrict the coverage of any claims that may be presented at any time in this application or in subsequent filings of this application. No disclaimer is intended. The following examples are being provided for nothing more than merely illustrative purposes. It is contemplated that the various teachings herein may be arranged and applied in numerous other ways. It is also contemplated that some variations may omit certain features referred to in the below examples. Therefore, none of the aspects or features referred to below should be deemed critical unless otherwise explicitly indicated as such at a later date by the inventors or by a successor in interest to the inventors. If any claims are presented in this application or in subsequent filings related to this application that include additional features beyond those referred to below, those additional features shall not be presumed to have been added for any reason relating to patentability.

[0178] Example 1

[0179] A fluidic apparatus comprising: (a) a first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas; (b) a second layer defining: (i) a second chamber portion positioned under 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 interposed between the second chamber portion and the at least one fluid outlet channel, the filter being formed in a 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 thereby 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 permit the at leastone liquid to flow therethrough, the filter being configured to prevent the plurality of particles from flowing therethrough.

[0180] Example 2

[0181] The fluidic apparatus of Example 1, the filter comprising a plurality of filter channels.

[0182] Example 3

[0183] The fluidic apparatus of Example 2, each filter channel of the plurality of filter channels being shallower than the at least one fluid outlet channel.

[0184] Example 4

[0185] The fluidic apparatus of any of Examples 2 through 3, each filter channel of the plurality of filter channels having a depth substantially less than about 50 pm.

[0186] Example 5

[0187] The fluidic apparatus of Example 4, each filter channel of the plurality of filter channels having a depth of about 25 pm.

[0188] Example 6

[0189] The fluidic apparatus of Example 4, each filter channel of the plurality of filter channels having a depth ranging from about 10 pm to about 23 pm.

[0190] Example 7

[0191] The fluidic apparatus of any of Examples 2 through 6, each filter channel of the plurality of filter channels extending between a respective inner end positioned proximate to the second chamber portion, and a respective outer end positioned distal from the second chamber portion.

[0192] Example 8

[0193] The fluidic apparatus of Example 7, the filter further comprising a filter entrance extending between the inner ends of the plurality of filter channels.

[0194] Example 9

[0195] The fluidic apparatus of any of Examples 7 through 8, the filter further comprising a filter exit extending between the outer ends of the plurality of filter channels.

[0196] Example 10

[0197] The fluidic apparatus of any of Examples 1 through 9. the second chamber portion having a sidewall, the second layer defining at least one sidewall channel extending along the sidewall.

[0198] Example 11

[0199] The fluidic apparatus of Example 10, the at least one sidewall channel extending along the sidewall to the filter.

[0200] Example 12

[0201] The fluidic apparatus of Example 11, the at least one sidewall channel including a plurality of sidewall channels extending along the sidewall to the filter.

[0202] Example 13

[0203] The fluidic apparatus of any of Examples 10 through 12, the second chamber portion having a floor, the second layer defining a floor channel extending along the floor.

[0204] Example 14

[0205] The fluidic apparatus of Example 13, the at least one sidewall channel extending along the sidewall to the floor channel.

[0206] Example 15

[0207] The fluidic apparatus of any of Examples 10 through 14, the at least one sidewall channel having a width of about 500 pm.

[0208] Example 16

[0209] The fluidic apparatus of any of Examples 10 through 15, the at least one sidewall channel having a depth of about 100 pm.

[0210] Example 17

[0211] The fluidic apparatus of any of Examples 1 through 16, the second layer further defining 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.

[0212] Example 18

[0213] The fluidic apparatus of Example 17, the particle inlet channel having at least one cross dimension greater than a corresponding cross dimension of the at least one fluid outlet channel.

[0214] Example 19

[0215] The fluidic apparatus of any of Examples 17 through 18, the particle inlet channel having a width of about 500 pm.

[0216] Example 20

[0217] The fluidic apparatus of any of Examples 17 through 19, the particle inlet channel having a depth of about 250 pm.

[0218] Example 21

[0219] The fluidic apparatus of any of Examples 1 through 20, the first layer further defining a third chamber portion, the third chamber portion being configured to receive pressurized gas, the second layer further defining: (i) a fourth chamber portion positioned under 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 and fourth chamber portions, the at least one bridging channel being configured to provide a path for communication of the at least one liquid and the plurality’ of particles between the second and fourth chamber portions.

[0220] Example 22

[0221] The fluidic apparatus of Example 21, the at least one bridging channel having a width of about 500 pm.

[0222] Example 23

[0223] The fluidic apparatus of any of Examples 21 through 22, the at least one bridging channel having a depth of about 500 pm.

[0224] Example 24

[0225] The fluidic apparatus of any of Examples 1 through 23, the second layer further defining 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.

[0226] Example 25

[0227] The fluidic apparatus of Example 24, the particle outlet channel having at least one cross dimension greater than a corresponding cross dimension of the at least one fluid outlet channel.

[0228] Example 26

[0229] A fluidic apparatus comprising: (a) a first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas; (b) a second layer defining: (i) a second chamber portion positioned under 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 channel in fluid communication with the second chamber portion, and (iii) a plurality of filter channels interposed 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 thereby 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 permit the at least one liquid to flow therethrough, the plurality of filter channels being configured to prevent the plurality of particles from flowing therethrough.

[0230] Example 27

[0231] The fluidic apparatus of Example 26, each filter channel of the plurality of filter channels extending between a respective inner end positioned proximate to thesecond chamber portion, and a respective outer end positioned distal from the second chamber portion.

[0232] Example 28

[0233] The fluidic apparatus of Example 27, the second layer further defining a filter entrance extending between the inner ends of the plurality of filter channels.

[0234] Example 29

[0235] The fluidic apparatus of any of Examples 26 through 27, the second layer further defining a filter exit extending between the outer ends of the plurality of filter channels.

[0236] Example 30

[0237] A fluidic apparatus comprising: (a) a first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas; (b) a second layer defining: (i) a second chamber portion positioned under the first chamber portion, the second chamber portion having a sidewall, the second chamber portion being configured to receive at least one liquid and a plurality of particles, (ii) a fluid outlet channel in fluid communication with the second chamber portion, (iii) a filter interposed between the second chamber portion and the at least one fluid outlet channel, the filter being formed in a 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 thereby drive the at least one liquid out of the second chamber portion via the plurality of sidewall channels, through the filter and into the fluid outlet channel, the filter being configured to permit the at least one liquid to flow therethrough, the filter being configured to prevent the plurality of particles from flowing therethrough.

[0238] Example 31

[0239] A method of using an apparatus, the apparatus comprising: (a) a first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas; (b) a second layer defining: (i) a second chamber portion positionedunder 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 interposed 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.

[0240] Example 32

[0241] The method of Example 31, the at least one liquid comprising an RNA therapeutic.

[0242] Example 33

[0243] The method of any of Examples 31 through 32, the plurality’ of particles comprising a plurality of beads.

[0244] Example 34

[0245] The method of any of Examples 31 through 33, the plurality’ of particles comprising a plurality of purification particles, the act of treating the at least one liquid comprising purifying the at least one liquid with the plurality of purification particles.

[0246] Example 35

[0247] The method of Example 34, each of the plurality of purification particles being configured to separate mRNA from one or more components of a transcription reaction process.

[0248] Example 36

[0249] The method of Example 35, the one or more components including at least one of plasmid DNA or enzy mes.

[0250] Example 37

[0251] The method of any of Examples 35 through 36, each of the plurality' of purification particles being configured to separate the mRNA from the components ofthe transcription reaction process by selectively capturing the mRNA via a polyadenylated (poly A) tail using salt and water purification steps.

[0252] Example 38

[0253] The method of any of Examples 34 through 37, each of the plurality of purification particles comprising a rigid, polymeric resin support matrix.

[0254] Example 39

[0255] The method of Example 38, the rigid, polymeric resin support matrix comprising cross-linked poly(styrene-divinylbenzene).

[0256] Example 40

[0257] The method of any of Examples 34 through 39, each of the plurality' of purification particles having a polyhydroxyl surface coating.

[0258] Example 41

[0259] The method of any of Examples 34 through 40, each of the plurality' of purification particles having a surface functionalized with poly(dT).

[0260] Example 42

[0261] The method of any of Examples 31 through 41, each of the plurality' of particles having a predetermined particle size.

[0262] Example 43

[0263] The method of Example 42, the predetermined particle size being about 50 pm.

[0264] Example 44

[0265] The method of any of Examples 31 through 43, each of the plurality' of particles being porous.

[0266] Example 45

[0267] The method of any of Examples 31 through 44, further comprising deforming the elastic layer into the second chamber portion to thereby drive the at least one liquidout of the second chamber portion through the filter and into the at least one fluid outlet channel.

[0268] Example 46

[0269] The method of Example 45, the filter permitting the at least one liquid to flow therethrough during the act of deforming, the filter preventing the plurality of particles from flowing therethrough during the act of deforming.

[0270] Example 47

[0271] The method of any of Examples 31 through 46, the second layer further defining a particle inlet channel in fluid communication with the second chamber portion, the method further comprising delivering the plurality of particles to the second chamber portion via the particle inlet channel.

[0272] Example 48

[0273] The method of any of Examples 31 through 47, the first layer further defining a third chamber portion, the third chamber portion being configured to receive pressurized gas, the second layer further defining: (i) a fourth chamber portion positioned under 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 and fourth chamber portions; the method further comprising communicating the at least one liquid and the plurality of particles between the second and fourth chamber portions via the at least one bridging channel.

[0274] Example 49

[0275] The method of any of Examples 31 through 48, the second layer further defining a particle outlet channel in fluid communication with the second chamber portion, the method further comprising removing the plurality of particles from the second chamber portion via the particle outlet channel.

[0276] Example 50

[0277] A method of using an apparatus, the apparatus comprising: (a) a first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas; (b) a second layer defining: (i) a second chamber portion positioned under 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 channel in fluid communication with the second chamber portion, and (iii) a plurality of filter channels interposed 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 comprising treating the at least one liquid with the plurality of particles within the second chamber portion.

[0278] Example 51

[0279] The method of Example 50, the at least one liquid comprising an RNA therapeutic.

[0280] Example 52

[0281] The method of any of Examples 50 through 51, the plurality of particles comprising a plurality of beads.

[0282] Example 53

[0283] The method of any of Examples 50 through 52, the plurality of particles comprising a plurality' of purification particles, the act of treating the at least one liquid comprising purifying the at least one liquid with the plurality of purification particles.

[0284] Example 54

[0285] The method of Example 53, each of the plurality of purification particles being configured to separate mRNA from one or more components of a transcription reaction process.

[0286] Example 55

[0287] The method of Example 54, the one or more components including at least one of plasmid DNA or enzymes.

[0288] Example 56

[0289] The method of any of Examples 54 through 55, each of the plurality of purification particles being configured to separate the mRNA from the components of the transcription reaction process by selectively capturing the mRNA via a polyadenylated (poly A) tail using salt and water purification steps.

[0290] Example 57

[0291] The method of any of Examples 53 through 56, each of the plurality' of purification particles comprising a rigid, polymeric resin support matrix.

[0292] Example 58

[0293] The method of Example 57, the rigid, polymeric resin support matrix comprising cross-linked poly(styrene-divinylbenzene).

[0294] Example 59

[0295] The method of any of Examples 53 through 58, each of the plurality' of purification particles having a polyhydroxyl surface coating.

[0296] Example 60

[0297] The method of any of Examples 53 through 59, each of the plurality' of purification particles having a surface functionalized with poly(dT).

[0298] Example 61

[0299] The method of any of Examples 50 through 60, each of the plurality’ of particles having a predetermined particle size.

[0300] Example 62

[0301] The method of Example 61, the predetermined particle size being about 50 pm.

[0302] Example 63

[0303] The method of any of Examples 50 through 62, each of the plurality’ of particles being porous.

[0304] Example 64

[0305] The method of any of Examples 50 through 63, further comprising deforming the elastic layer into the second chamber portion to thereby drive the at least one liquid out of the second chamber portion through the plurality of filter channels and into the fluid outlet channel.

[0306] Example 65

[0307] The method of Example 64, the plurality of filter channels permitting the at least one liquid to flow therethrough during the act of deforming, the plurality of filter channels preventing the plurality of particles from flowing therethrough during the act of deforming.

[0308] Example 66

[0309] The method of any of Examples 50 through 65, the second layer further defining a particle inlet channel in fluid communication with the second chamber portion, the method further comprising delivering the plurality of particles to the second chamber portion via the particle inlet channel.

[0310] Example 67

[0311] The method of any of Examples 50 through 66, the first layer further defining a third chamber portion, the third chamber portion being configured to receive pressurized gas, the second layer further defining: (i) a fourth chamber portion positioned under 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 and fourth chamber portions; the method further comprising communicating the at least one liquid and the plurality of particles between the second and fourth chamber portions via the at least one bridging channel.

[0312] Example 68

[0313] The method of any of Examples 50 through 67, the second layer further defining a particle outlet channel in fluid communication with the second chamber portion, the method further comprising removing the plurality of particles from the second chamber portion via the particle outlet channel.

[0314] Example 69

[0315] A method of using an apparatus, the apparatus comprising: (a) a first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas; (b) a second layer defining: (i) a second chamber portion positioned under the first chamber portion, the second chamber portion having a sidewall, the second chamber portion being configured to receive at least one liquid and a plurality of particles, (ii) a fluid outlet channel in fluid communication with the second chamber portion, (iii) a filter interposed between the second chamber portion and the at least one fluid outlet channel, 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 comprising treating the at least one liquid with the plurality of particles within the second chamber portion.

[0316] Example 70

[0317] The method of Example 69, the at least one liquid comprising an RNA therapeutic.

[0318] Example 71

[0319] The method of any of Examples 69 through 70, the plurality of particles comprising a plurality of beads.

[0320] Example 72

[0321] The method of any of Examples 69 through 71, the plurality of particles comprising a plurality of purification particles, the act of treating the at least one liquid comprising purifying the at least one liquid with the plurality of purification particles.

[0322] Example 73

[0323] The method of Example 72, each of the plurality of purification particles being configured to separate mRNA from components of a transcription reaction process.

[0324] Example 74

[0325] The method of Example 73, the components including at least one of plasmid DNA or enzymes.

[0326] Example 75

[0327] The method of any of Examples 73 through 74, each of the plurality of purification particles being configured to separate the mRNA from the components of the transcription reaction process by selectively capturing the mRNA via a polyadenylated (poly A) tail using salt and water purification steps.

[0328] Example 76

[0329] The method of any of Examples 72 through 75, each of the plurality' of purification particles comprising a rigid, polymeric resin support matrix.

[0330] Example 77

[0331] The method of Example 76, the rigid, polymeric resin support matrix comprising cross-linked poly(styrene-divinylbenzene).

[0332] Example 78

[0333] The method of any of Examples 72 through 77, each of the plurality' of purification particles having a polyhydroxyl surface coating.

[0334] Example 79

[0335] The method of any of Examples 72 through 78, each of the plurality’ of purification particles having a surface functionalized with poly(dT).

[0336] Example 80

[0337] The method of any of Examples 69 through 79, each of the plurality’ of particles having a predetermined particle size.

[0338] Example 81

[0339] The method of Example 80, the predetermined particle size being about 50 pm.

[0340] Example 82

[0341] The method of any of Examples 69 through 81, each of the plurality of particles being porous.

[0342] Example 83

[0343] The method of any of Examples 69 through 82, further comprising deforming the elastic layer into the second chamber portion to thereby drive the at least one liquid out of the second chamber portion via the plurality of sidewall channels, through the filter and into the fluid outlet channel.

[0344] Example 84

[0345] The method of Example 83, the filter permitting the at least one liquid to flow therethrough during the act of deforming, the filter preventing the plurality of particles from flowing therethrough during the act of deforming.

[0346] Example 85

[0347] The method of any of Examples 69 through 84, the second layer further defining a particle inlet channel in fluid communication with the second chamber portion, the method further comprising delivering the plurality of particles to the second chamber portion via the particle inlet channel.

[0348] Example 86

[0349] The method of any of Examples 69 through 85, the first layer further defining a third chamber portion, the third chamber portion being configured to receive pressurized gas, the second layer further defining: (i) a fourth chamber portion positioned under 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 and fourth chamber portions; the method further comprising communicating the at least one liquid and the plurality of particles between the second and fourth chamber portions via the at least one bridging channel.

[0350] Example 87

[0351] The method of any of Examples 69 through 86, the second layer further defining a particle outlet channel in fluid communication with the second chamber portion, the method further comprising removing the plurality of particles from the second chamber portion via the particle outlet channel.

[0352] Example 88

[0353] A fluidic apparatus comprising: (a) a first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas; (b) a second layer defining: (i) a second chamber portion positioned under 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 thereby drive the at least one liquid out of the second chamber portion; and (d) a filter interposed 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 permit the at least one liquid to flow therethrough, the filter being configured to prevent the plurality of particles from flowing therethrough.

[0354] Example 89

[0355] The fluidic apparatus of Example 88, the filter comprising at least one filter channel.

[0356] Example 90

[0357] The fluidic apparatus of Example 89, the at least one filter channel being shallower than the at least one fluid channel.

[0358] Example 91

[0359] The fluidic apparatus of any of Examples 89 through 90, the at least one filter channel having a depth substantially less than about 50 pm.

[0360] Example 92

[0361] The fluidic apparatus of Example 91, the at least one filter channel having a depth of about 25 pm.

[0362] Example 93

[0363] The fluidic apparatus of Example 91, the at least one filter channel having a depth of about 10 pm.

[0364] Example 94

[0365] The fluidic apparatus of any of Examples 88 through 93, the filter being formed in at least one surface of the second layer.

[0366] Example 95

[0367] The fluidic apparatus of any of Examples 88 through 93, the filter being formed in at least one surface of the elastic layer.

[0368] Example 96

[0369] The fluidic apparatus of Example 95, the elastic layer including an upper membrane and a lower membrane.

[0370] Example 97

[0371] The fluidic apparatus of Example 96, the filter including an aperture extending through the upper membrane.

[0372] Example 98

[0373] The fluidic apparatus of Example 97, the aperture defining a filter channel between the first layer and the lower membrane.

[0374] Example 99

[0375] The fluidic apparatus of any of Examples 96 through 98, the filter including first and second slots extending through the lower membrane.

[0376] Example 100

[0377] The fluidic apparatus of Example 99, the first and second slots defining a filter entrance and a filter exit, respectively.

[0378] Example 101

[0379] The fluidic apparatus of Example 100, the filter entrance being in fluid communication with the second chamber portion, the filter exit being in fluid communication with the at least one fluid channel.

[0380] Example 102

[0381] The fluidic apparatus of any of Examples 96 through 101, the upper membrane having a thickness of about 25 pm.

[0382] Example 103

[0383] The fluidic apparatus of any of Examples 96 through 101, the upper membrane having a thickness of about 10 pm.

[0384] Example 104

[0385] The fluidic apparatus of any of Examples 88 through 103, the at least one fluid channel including at least one fluid inlet channel.

[0386] Example 105

[0387] The fluidic apparatus of any of Examples 88 through 104, the at least one fluid channel including at least one fluid outlet channel.

[0388] Example 106

[0389] A fluidic apparatus comprising: (a) a first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas; (b) a second layer defining: (i) a second chamber portion positioned under 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 thereby drive the at least one liquid out of the second chamber portion; and (d) at least one filter channel interposed 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 permit the at leastone liquid to flow therethrough, the at least one filter channel being configured to prevent the plurality of particles from flowing therethrough.

[0390] Example 107

[0391] The fluidic apparatus of Example 106, the at least one filter channel being formed in at least one surface of the elastic layer.

[0392] Example 108

[0393] The fluidic apparatus of Example 107, the elastic layer including an upper membrane and a lower membrane.

[0394] Example 109

[0395] The fluidic apparatus of Example 108, the at least one filter channel being defined by an aperture extending through the upper membrane.

[0396] Example 110

[0397] A fluidic apparatus comprising: (a) a first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas; (b) a second layer defining: (i) a second chamber portion positioned under the first chamber portion, the second chamber portion having a sidewall, the second chamber portion being configured to receive at least one liquid and a plurality7of particles, (ii) a fluid channel in fluid communication with the second chamber portion, 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 thereby drive the at least one liquid out of the second chamber portion via the plurality of sidewall channels; and (d) a filter interposed 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 permit the at least one liquid to flow therethrough, the filter being configured to prevent the plurality of particles from flowing therethrough, the plurality of sidewall channels extending along the sidewall to the filter.

[0398] Example 111

[0399] A method of using an apparatus, the apparatus comprising: (a) a first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas; (b) a second layer defining: (i) a second chamber portion positioned under 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 interposed 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.

[0400] Example 112

[0401] The method of Example 111, the at least one liquid comprising an RNA therapeutic.

[0402] Example 113

[0403] The method of any of Examples 111 through 112, the plurality of particles comprising a plurality of beads.

[0404] Example 114

[0405] The method of any of Examples 111 through 113, the plurality of particles comprising a plurality' of purification particles, the act of treating the at least one liquid comprising purifying the at least one liquid with the plurality of purification particles.

[0406] Example 115

[0407] The method of Example 114, each of the plurality of purification particles being configured to separate mRNA from one or more components of a transcription reaction process.

[0408] Example 116

[0409] The method of Example 115, the one or more components including at least one of plasmid DNA or enzymes.

[0410] Example 117

[0411] The method of any of Examples 115 through 116, each of the plurality of purification particles being configured to separate the mRNA from the components of the transcription reaction process by selectively capturing the mRNA via a polyadenylated (poly A) tail using salt and water purification steps.

[0412] Example 118

[0413] The method of any of Examples 114 through 117, each of the plurality of purification particles comprising a rigid, polymeric resin support matrix.

[0414] Example 119

[0415] The method of Example 118, the rigid, polymeric resin support matrix comprising cross-linked poly(styrene-divinylbenzene).

[0416] Example 120

[0417] The method of any of Examples 114 through 119, each of the plurality of purification particles having a polyhydroxyl surface coating.

[0418] Example 121

[0419] The method of any of Examples 114 through 120, each of the plurality of purification particles having a surface functionalized with poly(dT).

[0420] Example 122

[0421] The method of any of Examples 111 through 121, each of the plurality of particles having a predetermined particle size.

[0422] Example 123

[0423] The method of Example 122, the predetermined particle size being about 50 pm.

[0424] Example 124

[0425] The method of any of Examples 111 through 123, each of the plurality of particles being porous.

[0426] Example 125

[0427] The method of any of Examples 111 through 124, further comprising deforming the elastic layer into the second chamber portion to thereby drive the at least one liquid out of the second chamber portion through the filter and into the at least one fluid channel.

[0428] Example 126

[0429] The method of Example 125, the filter permitting the at least one liquid to flow therethrough during the act of deforming, the filter preventing the plurality of particles from flowing therethrough during the act of deforming.

[0430] Example 127

[0431] The method of any of Examples 111 through 126, the second layer further defining a particle inlet channel in fluid communication with the second chamber portion, the method further comprising delivering the plurality of particles to the second chamber portion via the particle inlet channel.

[0432] Example 128

[0433] The method of any of Examples 111 through 127, the first layer further defining a third chamber portion, the third chamber portion being configured to receive pressurized gas, the second layer further defining: (i) a fourth chamber portion positioned under 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 and fourth chamber portions; the method further comprising communicating the at least one liquid and the plurality of particles between the second and fourth chamber portions via the at least one bridging channel.

[0434] Example 129

[0435] The method of any of Examples 111 through 128, the second layer further defining a particle outlet channel in fluid communication with the second chamber portion, the method further comprising removing the plurality of particles from the second chamber portion via the particle outlet channel.

[0436] Example 130

[0437] A method of using an apparatus, the apparatus comprising: (a) a first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas; (b) a second layer defining: (i) a second chamber portion positioned under 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 interposed 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.

[0438] Example 131

[0439] The method of Example 130, the at least one liquid comprising an RNA therapeutic.

[0440] Example 132

[0441] The method of any of Examples 130 through 131, the plurality of particles comprising a plurality of beads.

[0442] Example 133

[0443] The method of any of Examples 130 through 132, the plurality of particles comprising a plurality of purification particles, the act of treating the at least one liquid comprising purifying the at least one liquid with the plurality of purification particles.

[0444] Example 134

[0445] The method of Example 133. each of the plurality of purification particles being configured to separate mRNA from one or more components of a transcription reaction process.

[0446] Example 135

[0447] The method of Example 134, the one or more components including at least one of plasmid DNA or enzy mes.

[0448] Example 136

[0449] The method of any of Examples 134 through 135, each of the plurality of purification particles being configured to separate the mRNA from the components of the transcription reaction process by selectively capturing the mRNA via a polyadenylated (poly A) tail using salt and water purification steps.

[0450] Example 137

[0451] The method of any of Examples 133 through 136, each of the plurality of purification particles comprising a rigid, polymeric resin support matrix.

[0452] Example 138

[0453] The method of Example 137. the rigid, polymeric resin support matrix comprising cross-linked poly(styrene-divinylbenzene).

[0454] Example 139

[0455] The method of any of Examples 133 through 138, each of the plurality of purification particles having a polyhydroxyl surface coating.

[0456] Example 140

[0457] The method of any of Examples 133 through 139, each of the plurality of purification particles having a surface functionalized with poly(dT).

[0458] Example 141

[0459] The method of any of Examples 130 through 140, each of the plurality of particles having a predetermined particle size.

[0460] Example 142

[0461] The method of Example 141, the predetermined particle size being about 50 pm.

[0462] Example 143

[0463] The method of any of Examples 130 through 142, each of the plurality’ of particles being porous.

[0464] Example 144

[0465] The method of any of Examples 130 through 143, further comprising deforming the elastic layer into the second chamber portion to thereby drive the at least one liquid out of the second chamber portion through the at least one filter channel and into the fluid channel.

[0466] Example 145

[0467] The method of Example 144, the at least one filter channel permitting the at least one liquid to flow therethrough during the act of deforming, the at least one filter channel preventing the plurality of particles from flowing therethrough during the act of deforming.

[0468] Example 146

[0469] The method of any of Examples 130 through 145, the second layer further defining a particle inlet channel in fluid communication with the second chamber portion, the method further comprising delivering the plurality of particles to the second chamber portion via the particle inlet channel.

[0470] Example 147

[0471] The method of any of Examples 130 through 146. the first layer further defining a third chamber portion, the third chamber portion being configured to receive pressurized gas, the second layer further defining: (i) a fourth chamber portion positioned under 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 and fourth chamber portions; the method further comprising communicating the at least one liquid and the plurality of particles between the second and fourth chamber portions via the at least one bridging channel.

[0472] Example 148

[0473] The method of any of Examples 130 through 147, the second layer further defining a particle outlet channel in fluid communication with the second chamber portion, the method further comprising removing the plurality of particles from the second chamber portion via the particle outlet channel.

[0474] Example 149

[0475] A method of using an apparatus, the apparatus comprising: (a) a first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas; (b) a second layer defining: (i) a second chamber portion positioned under the first chamber portion, the second chamber portion having a sidewall, the second chamber portion being 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 sidewall; (c) an elastic layer disposed between the first layer and the second layer; and (d) a filter interposed 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 comprising treating the at least one liquid with the plurality of particles within the second chamber portion.

[0476] Example 150

[0477] The method of Example 149, the at least one liquid comprising an RNA therapeutic.

[0478] Example 151

[0479] The method of any of Examples 149 through 150. the plurality of particles comprising a plurality of beads.

[0480] Example 152

[0481] The method of any of Examples 149 through 151. the plurality of particles comprising a plurality of purification particles, the act of treating the at least one liquid comprising purifying the at least one liquid with the plurality of purification particles.

[0482] Example 153

[0483] The method of Example 152, each of the plurality of purification particles being configured to separate mRNA from components of a transcription reaction process.

[0484] Example 154

[0485] The method of Example 153, the components including at least one of plasmid DNA or enzymes.

[0486] Example 155

[0487] The method of any of Examples 153 through 154, each of the plurality of purification particles being configured to separate the mRNA from the components of the transcription reaction process by selectively capturing the mRNA via a polyadenylated (poly A) tail using salt and water purification steps.

[0488] Example 156

[0489] The method of any of Examples 152 through 155, each of the plurality of purification particles comprising a rigid, polymeric resin support matrix.

[0490] Example 157

[0491] The method of Example 156, the rigid, polymeric resin support matrix comprising cross-linked poly(styrene-divinylbenzene).

[0492] Example 158

[0493] The method of any of Examples 152 through 157, each of the plurality of purification particles having a polyhydroxyl surface coating.

[0494] Example 159

[0495] The method of any of Examples 152 through 158, each of the plurality of purification particles having a surface functionalized with poly(dT).

[0496] Example 160

[0497] The method of any of Examples 149 through 159, each of the plurality’ of particles having a predetermined particle size.

[0498] Example 161

[0499] The method of Example 160, the predetermined particle size being about 50 pm.

[0500] Example 162

[0501] The method of any of Examples 149 through 161, each of the plurality of particles being porous.

[0502] Example 163

[0503] The method of any of Examples 149 through 162, further comprising deforming the elastic layer into the second chamber portion to thereby drive the at least one liquid out of the second chamber portion via the plurality of sidewall channels, through the filter and into the fluid channel.

[0504] Example 164

[0505] The method of Example 163, the filter permitting the at least one liquid to flow therethrough during the act of deforming, the filter preventing the plurality of particles from flowing therethrough during the act of deforming.

[0506] Example 165

[0507] The method of any of Examples 149 through 164, the second layer further defining a particle inlet channel in fluid communication with the second chamber portion, the method further comprising delivering the plurality of particles to the second chamber portion via the particle inlet channel.

[0508] Example 166

[0509] The method of any of Examples 149 through 165, the first layer further defining a third chamber portion, the third chamber portion being configured to receive pressurized gas, the second layer further defining: (i) a fourth chamber portion positioned under 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 and fourth chamber portions; the method further comprising communicating the at least one liquid and the plurality of particles between the second and fourth chamber portions via the at least one bridging channel.

[0510] Example 167

[0511] The method of any of Examples 149 through 166, the second layer further defining a particle outlet channel in fluid communication with the second chamber portion, the method further comprising removing the plurality of particles from the second chamber portion via the particle outlet channel.

[0512] Example 168

[0513] The fluidic apparatus of any of Examples 2-25, each filter channel of the plurality of filter channels having at least one cross dimension that is substantially greater than about 50 pm.

[0514] Example 169

[0515] The fluidic apparatus of Example 168, the at least one cross dimension being substantially less than about 250 pm.

[0516] Example 170

[0517] The fluidic apparatus of any of Examples 168-169, the at least one cross dimension including at least one of a width or a depth.

[0518] Example 171

[0519] The fluidic apparatus of Example 170, the at least one cross dimension including both the width and the depth.

[0520] Example 172

[0521] The fluidic apparatus of any of Examples 170-171, the depth being greater than or equal to about 100 pm.

[0522] Example 173

[0523] The fluidic apparatus of any of Examples 170-172, the width ranging from about 100 pm to about 400 pm.

[0524] Example 174

[0525] The fluidic apparatus of Example 173, the width being about 200 pm.

[0526] Example 175

[0527] The fluidic apparatus of any of Examples 1-25 or 168-174. the second layer defining at least one backflush channel in fluid communication with the at least one fluid outlet channel, the at least one backflush channel being configured to direct at least one backflush fluid through the filter and into the second chamber portion.

[0528] Example 176

[0529] The fluidic apparatus of any of Examples 26-29, the second layer defining at least one backflush channel in fluid communication with the plurality of filter channels, the at least one backflush channel being configured to direct at least one backflush fluid through the plurality of filter channels and into the second chamber portion.

[0530] Example 177

[0531] The method of any of Examples 31-49, the second layer further defining at least one backflush channel in fluid communication with the at least one fluid outlet channel, the method further comprising directing at least one backflush fluid from the at least one backflush channel through the filter and into the second chamber portion.

[0532] Example 178

[0533] The method of any of Examples 50-68, the second layer defining at least one backflush channel in fluid communication with the plurality of filter channels, the method further comprising directing at least one backflush fluid from the at least one backflush channel through the plurality of filter channels and into the second chamber portion.

[0534] Example 179

[0535] The method of any of Examples 69-87, the second layer defining at least one backflush channel in fluid communication with the at least one fluid outlet channel, the method further comprising directing at least one backflush fluid from the at least one backflush channel through the filter and into the second chamber portion.

[0536] Example 180

[0537] The fluidic apparatus of any of Examples 89-105, the at least one filter channel having at least one cross dimension that is substantially greater than about 50 pm.

[0538] Example 181

[0539] The fluidic apparatus of Example 180, the at least one cross dimension being substantially less than about 250 pm.

[0540] Example 182

[0541] The fluidic apparatus of any of Examples 180-181. the at least one cross dimension including at least one of a width or a depth.

[0542] Example 183

[0543] The fluidic apparatus of Example 182, the at least one cross dimension including both the width and the depth.

[0544] Example 184

[0545] The fluidic apparatus of any of Examples 182-183, the depth being greater than or equal to about 100 pm.

[0546] Example 185

[0547] The fluidic apparatus of any of Examples 182-184, the width ranging from about 100 pm to about 400 pm.

[0548] Example 186

[0549] The fluidic apparatus of Example 185, the width being about 200 pm.

[0550] Example 187

[0551] The fluidic apparatus of any of Examples 88-105 or 180-186, the second layer defining at least one backflush channel in fluid communication with the at least one fluid outlet channel, the at least one backflush channel being configured to direct at least one backflush fluid through the filter and into the second chamber portion.

[0552] Example 188

[0553] The fluidic apparatus of any of Examples 106-109, the second layer defining at least one backflush channel in fluid communication with the at least one filter channel, the at least one backflush channel being configured to direct at least one backflush fluid through the at least one filter channel and into the second chamber portion.

[0554] Example 189

[0555] The fluidic apparatus of Example 110, the second layer defining at least one backflush channel in fluid communication with the at least one fluid channel, the at least one backflush channel being configured to direct at least one backflush fluid through the filter and into the second chamber portion.

[0556] Example 190

[0557] The method of any of Examples 111-129, the second layer defining at least one backflush channel in fluid communication with the at least one fluid channel, the method further comprising directing at least one backflush fluid from the at least one backflush channel through the filter and into the second chamber portion.

[0558] Example 191

[0559] The method of any of Examples 130-148, the second layer defining at least one backflush channel in fluid communication with the at least one filter channel, the method further comprising directing 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.

[0560] Example 192

[0561] The method of any of Examples 149-167, the second layer defining at least one backflush channel in fluid communication with the at least one fluid channel, the method further comprising directing at least one backflush fluid from the at least one backflush channel through the filter and into the second chamber portion.

[0562] VII. Miscellaneous

[0563] The foregoing description is provided to enable a person skilled in the art to practice the various configurations described herein. While the subject technology has been particularly described with reference to the various figures and configurations, it should be understood that these are for illustration purposes only and should not be taken as limiting the scope of the subject technology.

[0564] There may be many other ways to implement the subj ect technology7. V arious functions and elements described herein may be partitioned differently from those shown without departing from the scope of the subject technology. Various modifications to these implementations may be readily apparent to those skilled in the art, and generic principles defined herein may be applied to other implementations. Thus, many changes and modifications may be made to the subject technology, by one having ordinary skill in the art, without departing from the scope of the subject technology. For instance, different numbers of a given module or unit may be employed, a different type or types of a given module or unit may be employed, a given module or unit may be added, or a given module or unit may be omitted.

[0565] Some versions of the examples described herein may be implemented using a processor, which may be part of a computer sy stem and communicate with a number of peripheral devices via bus subsystem. Versions of the examples described herein that are implemented using a computer system may be implemented using a general- purpose computer that is programmed to perform the methods described herein. Alternatively, versions of the examples described herein that are implemented using a computer system may be implemented using a specific-purpose computer that is constructed with hardware arranged to perform the methods described herein. Versions of the examples described herein may also be implemented using a combination of at least one general-purpose computer and at least one specific-purpose computer.

[0566] In versions implemented using a computer system, each processor may include a central processing unit (CPU) of a computer system, a microprocessor, an application-specific integrated circuit (ASIC), other kinds of hardware components, and combinations thereof. A computer system may include more than one type of processor. The peripheral devices of a computer system may include a storagesubsystem including, for example, memory devices and a file storage subsystem, user interface input devices, user interface output devices, and a network interface subsystem. The input and output devices may allow user interaction with the computer system. The network interface subsystem may provide an interface to outside networks, including an interface to corresponding interface devices in other computer systems. User interface input devices may include a keyboard; pointing devices such as a mouse, trackball, touchpad, or graphics tablet; a scanner; a touch screen incorporated into the display; audio input devices such as voice recognition systems and microphones; and other ty pes of input devices. In general, use of the term "input device" is intended to include all possible types of devices and ways to input information into computer system.

[0567] In versions implemented using a computer system, a user interface output device may include a display subsystem, a printer, a fax machine, or non-visual displays such as audio output devices. The display subsystem may include a cathode ray tube (CRT), a flat-panel device such as a liquid cry stal 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 audio output devices. In general, use of the term "output device" is intended to include all possible types of devices and ways to output information from computer system to the user or to another machine or computer system.

[0568] In versions implemented using a computer system, a storage subsystem may store programming and data constructs that provide the functionality of some or all of the modules and methods described herein. These software modules may be generally executed by the processor of the computer system alone or in combination with other processors. Memory7used in the storage subsystem may include a number of memories including a main random-access memory (RAM) for storage of instructions and data during program execution and a read only memory (ROM) in which fixed instructions are stored. A file storage subsystem may provide persistent storage for program and data files, and may7include a hard disk drive, a floppy7disk drive along with associated removable media, a CD-ROM drive, an optical drive, or removable media cartridges. The modules implementing the functionality of certain implementations may be storedby file storage subsystem in the storage subsystem, or in other machines accessible by the processor.

[0569] In versions implemented using a computer system, the computer system itself may be of varying types including a personal computer, a portable computer, a workstation, a computer terminal, a network computer, a television, a mainframe, a server farm, a widely-distributed set of loosely networked computers, or any other data processing system or user device. Due to the ever-changing nature of computers and networks, the example of the computer system described herein is intended only as a specific example for purposes of illustrating the technology disclosed. Many other configurations of a computer system are possible having more or fewer components than the computer system described herein.

[0570] As an article of manufacture, rather than a method, a non-transitoiy computer readable medium (CRM) may be loaded with program instructions executable by a processor. The program instructions when executed, implement one or more of the computer-implemented methods described above. Alternatively, the program instructions may be loaded on a non-transitory CRM and, when combined with appropriate hardware, become a component of one or more of the computer- implemented systems that practice the methods disclosed.

[0571] Underlined and / or italicized headings and subheadings are used for convenience only, do not limit the subject technology, and are not referred to in connection with the interpretation of the description of the subject technology. All structural and functional equivalents to the elements of the various implementations described throughout this disclosure that are known or later come to be known to those of ordinary skill in the art are expressly incorporated herein by reference and intended to be encompassed by the subject technology. Moreover, nothing disclosed herein is intended to be dedicated to the public regardless of whether such disclosure is explicitly recited in the above description.

[0572] It should be appreciated that all combinations of the foregoing concepts and additional concepts discussed in greater detail below (provided such concepts are not mutually inconsistent) are contemplated as being part of the inventive subject matterdisclosed herein. In particular, all combinations of claimed subject matter appearing at the end of this disclosure are contemplated as being part of the inventive subject matter disclosed herein.

Claims

WHAT IS CLAIMED IS:

1. A fluidic apparatus comprising:(a) a first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas;(b) a second layer defining:(i) a second chamber portion positioned under 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 interposed between the second chamber portion and the at least one fluid outlet channel, the filter being formed in a 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 thereby 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 permit the at least one liquid to flow therethrough, the filter being configured to prevent the plurality of particles from flowing therethrough.

2. The fluidic apparatus of claim 1 , the filter comprising a plurality of filter channels.

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

4. The fluidic apparatus of any of claims 2 through 3, each filter channel of the plurality of filter channels having a depth substantially less than about 50 pm.

5. The fluidic apparatus of claim 4, each filter channel of the plurality of filter channels having a depth of about 25 pm.

6. The fluidic apparatus of claim 4. each filter channel of the plurality' of filter channels having a depth ranging from about 10 pm to about 23 pm.

7. The fluidic apparatus of any of claims 2 through 6, each filter channel of the plurality of filter channels extending between a respective inner end positioned proximate to the second chamber portion, and a respective outer end positioned distal from the second chamber portion.

8. The fluidic apparatus of claim 7, the filter further comprising a filter entrance extending between the inner ends of the plurality of filter channels.

9. The fluidic apparatus of any of claims 7 through 8, the filter further comprising a filter exit extending between the outer ends of the plurality7of filter channels.

10. The fluidic apparatus of any of claims 1 through 9, the second chamber portion having a sidewall, the second layer defining at least one sidewall channel extending along the sidewall.

11. The fluidic apparatus of claim 10, the at least one sidewall channel extending along the sidewall to the filter.

12. The fluidic apparatus of claim 11, the at least one sidewall channel including a plurality of sidewall channels extending along the sidewall to the filter.

13. The fluidic apparatus of any of claims 10 through 12, the second chamber portion having a floor, the second layer defining a floor channel extending along the floor.

14. The fluidic apparatus of claim 13, the at least one sidewall channel extending along the sidewall to the floor channel.

15. The fluidic apparatus of any of claims 10 through 14, the at least one sidewall channel having a width of about 500 pm.

16. The fluidic apparatus of any of claims 10 through 15, the at least one sidewall channel having a depth of about 100 pm.

17. The fluidic apparatus of any of claims 1 through 16, the second layer further defining 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 fluidic apparatus of claim 17. the particle inlet channel having at least one cross dimension greater than a corresponding cross dimension of the at least one fluid outlet channel.

19. The fluidic apparatus of any of claims 17 through 18, the particle inlet channel having a width of about 500 pm.

20. The fluidic apparatus of any of claims 17 through 19, the particle inlet channel having a depth of about 250 pm.

21. The fluidic apparatus of any of claims 1 through 20, the first layer further defining a third chamber portion, the third chamber portion being configured to receive pressurized gas, the second layer further defining:(i) a fourth chamber portion positioned under 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 and fourth chamber portions, the at least one bridging channel being configured to provide a path for communication of the at least one liquid and the plurality of particles between the second and fourth chamber portions.

22. The fluidic apparatus of claim 21 , the at least one bridging channel having a width of about 500 pm.

23. The fluidic apparatus of any of claims 21 through 22, the at least one bridging channel having a depth of about 500 pm.

24. The fluidic apparatus of any of claims 1 through 23, the second layer further defining 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 fluidic apparatus of claim 24, the particle outlet channel having at least one cross dimension greater than a corresponding cross dimension of the at least one fluid outlet channel.

26. A fluidic apparatus comprising:(a) a first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas;(b) a second layer defining:(i) a second chamber portion positioned under 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 channel in fluid communication with the second chamber portion, and(iii) a plurality of filter channels interposed 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 thereby 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 permit the at least one liquid to flow therethrough, the plurality of filter channels being configured to prevent the plurality' of particles from flowing therethrough.

27. The fluidic apparatus of claim 26, each filter channel of the plurality of filter channels extending between a respective inner end positioned proximate to the second chamber portion, and a respective outer end positioned distal from the second chamber portion.

28. The fluidic apparatus of claim 27, the second layer further defining a filter entrance extending between the inner ends of the plurality of filter channels.

29. The fluidic apparatus of any of claims 26 through 27, the second layer further defining a filter exit extending between the outer ends of the plurality of filter channels.

30. A fluidic apparatus comprising:(a) a first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas;(b) a second layer defining:(i) a second chamber portion positioned under the first chamber portion, the second chamber portion having a sidewall, the second chamber portion being configured to receive at least one liquid and a plurality of particles,(ii) a fluid outlet channel in fluid communication with the second chamber portion,(iii) a filter interposed between the second chamber portion and the at least one fluid outlet channel, the filter being formed in a 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 thereby drive the at least one liquid out of the second chamber portion via the plurality of sidewall channels, through the filter and into the fluid outlet channel, the filter being configured to permit the at least one liquid to flow therethrough, the filter being configured to prevent the plurality of particles from flowing therethrough.

31. A method of using an apparatus, the apparatus comprising:(a) a first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas;(b) a second layer defining:(i) a second chamber portion positioned under 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 interposed 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.

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

33. The method of any of claims 31 through 32, the plurality7of particles comprising a plurality of beads.

34. The method of any of claims 31 through 33, the plurality of particles comprising a plurality of purification particles, the act of treating the at least one liquid comprising purify ing the at least one liquid with the plurality7of purification particles.

35. The method of claim 34. each of the plurality of purification particles being configured to separate mRNA from one or more components of a transcription reaction process.

36. The method of claim 35, the one or more components including at least one of plasmid DNA or enzymes.

37. The method of any of claims 35 through 36, each of the plurality of purification particles being configured to separate the mRNA from the components of the transcription reaction process by selectively capturing the mRNA via a polyadenylated (poly A) tail using salt and water purification steps.

38. The method of any of claims 34 through 37, each of the plurality of purification particles comprising a rigid, polymeric resin support matrix.

39. The method of claim 38, the rigid, polymeric resin support matrix comprising cross-linked poly(styrene-divinylbenzene).

40. The method of any of claims 34 through 39, each of the lurality of purification particles having a polyhydroxyl surface coating.

41. The method of any of claims 34 through 40, each of the plurality of purification particles having a surface functionalized with poly(dT).

42. The method of any of claims 31 through 41, each of the plurality of particles having a predetermined particle size.

43. The method of claim 42, the predetermined particle size being about 50 pm.

44. The method of any of claims 31 through 43, each of the plurality of particles being porous.

45. The method of any of claims 31 through 44, further comprising deforming the elastic layer into the second chamber portion to thereby 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, the filter permitting the at least one liquid to flow therethrough during the act of deforming, the filter preventing the plurality of particles from flowing therethrough during the act of deforming.

47. The method of any of claims 31 through 46, the second layer further defining a particle inlet channel in fluid communication with the second chamber portion, the method further comprising delivering the plurality of particles to the second chamber portion via the particle inlet channel.

48. The method of any of claims 31 through 47, the first layer further defining a third chamber portion, the third chamber portion being configured to receive pressurized gas, the second layer further defining:(i) a fourth chamber portion positioned under 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 and fourth chamber portions; the method further comprising communicating the at least one liquid and the plurality of particles between the second and fourth chamber portions via the at least one bridging channel.

49. The method of any of claims 31 through 48, the second layer further defining a particle outlet channel in fluid communication with the second chamber portion, the method further comprising 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 defining a first chamber portion, the first chamber portion being configured to receive pressurized gas;(b) a second layer defining:(i) a second chamber portion positioned under 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 channel in fluid communication with the second chamber portion, and(iii) a plurality of filter channels interposed 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 comprising treating the at least one liquid with the plurality of particles within the second chamber portion.

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

52. The method of any of claims 50 through 51, the plurality7of particles comprising a plurality' of beads.

53. The method of any of claims 50 through 52, the plurality of particles comprising a plurality of purification particles, the act of treating the at least one liquid comprising purifying the at least one liquid with the plurality7of purification particles.

54. The method of claim 53. each of the plurality of purification particles being configured to separate mRNA from one or more components of a transcription reaction process.

55. The method of claim 54, the one or more components including at least one of plasmid DNA or enzy mes.

56. The method of any of claims 54 through 55, each of the plurality7of purification particles being configured to separate the mRNA from the components of the transcription reaction process by selectively capturing the mRNA via a polyadenylated (poly A) tail using salt and water purification steps.

57. The method of any of claims 53 through 56, each of the plurality of purification particles comprising a rigid, polymeric resin support matrix.

58. The method of claim 57, the rigid, polymeric resin support matrix comprising cross-linked poly(styrene-divinylbenzene).

59. The method of any of claims 53 through 58, each of the lurality of purification particles having a polyhydroxyl surface coating.

60. The method of any of claims 53 through 59, each of the plurality of purification particles having a surface functionalized with poly(dT).

61. The method of any of claims 50 through 60, each of the plurality of particles having a predetermined particle size.

62. The method of claim 61, the predetermined particle size being about 50 pm.

63. The method of any of claims 50 through 62, each of the plurality of particles being porous.

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

65. The method of claim 64, the plurality of filter channels permitting the at least one liquid to flow therethrough during the act of deforming, the plurality of filter channels preventing the plurality of particles from flowing therethrough during the act of deforming.

66. The method of any of claims 50 through 65, the second layer further defining a particle inlet channel in fluid communication with the second chamber portion, the method further comprising delivering the plurality of particles to the second chamber portion via the particle inlet channel.

67. The method of any of claims 50 through 66, the first layer further defining a third chamber portion, the third chamber portion being configured to receive pressurized gas, the second layer further defining:(i) a fourth chamber portion positioned under 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 and fourth chamber portions; the method further comprising communicating the at least one liquid and the plurality of particles between the second and fourth chamber portions via the at least one bridging channel.

68. The method of any of claims 50 through 67, the second layer further defining a particle outlet channel in fluid communication with the second chamber portion, the method further comprising 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 defining a first chamber portion, the first chamber portion being configured to receive pressurized gas;(b) a second layer defining:(i) a second chamber portion positioned under the first chamber portion, the second chamber portion having a sidewall, the second chamber portion being configured to receive at least one liquid and a plurality of particles,(ii) a fluid outlet channel in fluid communication with the second chamber portion,(iii) a filter interposed between the second chamber portion and the at least one fluid outlet channel, 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 comprising treating the at least one liquid with the plurality of particles within the second chamber portion.

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

71. The method of any of claims 69 through 70, the plurality of particles comprising a plurality of beads.

72. The method of any of claims 69 through 71, the plurality of particles comprising a plurality of purification particles, the act of treating the at least one liquid comprising purifying the at least one liquid with the plurality of purification particles.

73. The method of claim 72, each of the plurality of purification particles being configured to separate mRNA from components of a transcription reaction process.

74. The method of claim 73, the components including at least one of plasmid DNA or enzy mes.

75. The method of any of claims 73 through 74, each of the plurality of purification particles being configured to separate the mRNA from the components ofthe transcription reaction process by selectively capturing the mRNA via a polyadenylated (poly A) tail using salt and water purification steps.

76. The method of any of claims 72 through 75, each of the plurality of purification particles comprising a rigid, polymeric resin support matrix.

77. The method of claim 76, the rigid, polymeric resin support matrix comprising cross-linked poly(styrene-divinylbenzene).

78. The method of any of claims 72 through 77, each of the plurality of purification particles having a polyhydroxyl surface coating.

79. The method of any of claims 72 through 78, each of the plurality of purification particles having a surface functionalized with poly(dT).

80. The method of any of claims 69 through 79, each of the plurality of particles having a predetermined particle size.

81. The method of claim 80, the predetermined particle size being about 50 pm.

82. The method of any of claims 69 through 81, each of the plurality of particles being porous.

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

84. The method of claim 83, the filter permitting the at least one liquid to flow therethrough during the act of deforming, the filter preventing the plurality of particles from flowing therethrough during the act of deforming.

85. The method of any of claims 69 through 84, the second layer further defining a particle inlet channel in fluid communication with the second chamber portion, the method further comprising delivering the plurality of particles to the second chamber portion via the particle inlet channel.

86. The method of any of claims 69 through 85, the first layer further defining a third chamber portion, the third chamber portion being configured to receive pressurized gas, the second layer further defining:(i) a fourth chamber portion positioned under 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 and fourth chamber portions; the method further comprising communicating the at least one liquid and the plurality of particles between the second and fourth chamber portions via the at least one bridging channel.

87. The method of any of claims 69 through 86, the second layer further defining a particle outlet channel in fluid communication with the second chamber portion, the method further comprising removing the plurality of particles from the second chamber portion via the particle outlet channel.

88. A fluidic apparatus comprising:(a) a first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas;(b) a second layer defining:(i) a second chamber portion positioned under 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 thereby drive the at least one liquid out of the second chamber portion; and(d) a filter interposed 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 permit the at least one liquid to flow therethrough, the filter being configured to prevent the plurality of particles from flowing therethrough.

89. The fluidic apparatus of claim 88, the filter comprising at least one filter channel.

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

91. The fluidic apparatus of any of claims 89 through 90, the at least one filter channel having a depth substantially less than about 50 pm.

92. The fluidic apparatus of claim 91, the at least one filter channel having a depth of about 25 pm.

93. The fluidic apparatus of claim 91, the at least one filter channel having a depth of about 10 pm.

94. The fluidic apparatus of any of claims 88 through 93, the filter being formed in at least one surface of the second layer.

95. The fluidic apparatus of any of claims 88 through 93, the filter being formed in at least one surface of the elastic layer.

96. The fluidic apparatus of claim 95, the elastic layer including an upper membrane and a lower membrane.

97. The fluidic apparatus of claim 96. the filter including an aperture extending through the upper membrane.

98. The fluidic apparatus of claim 97, the aperture defining a filter channel between the first layer and the lower membrane.

99. The fluidic apparatus of any of claims 96 through 98, the filter including first and second slots extending through the lower membrane.

100. The fluidic apparatus of claim 99, the first and second slots defining a filter entrance and a filter exit, respectively.

101. The fluidic apparatus of claim 100, the filter entrance being in fluid communication with the second chamber portion, the filter exit being in fluid communication with the at least one fluid channel.

102. The fluidic apparatus of any of claims 96 through 101, the upper membrane having a thickness of about 25 pm.

103. The fluidic apparatus of any of claims 96 through 101, the upper membrane having a thickness of about 10 pm.

104. The fluidic apparatus of any of claims 88 through 103, the at least one fluid channel including at least one fluid inlet channel.

105. The fluidic apparatus of any of claims 88 through 104, the at least one fluid channel including at least one fluid outlet channel.

106. A fluidic apparatus comprising:(a) a first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas;(b) a second layer defining:(i) a second chamber portion positioned under 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 thereby drive the at least one liquid out of the second chamber portion; and(d) at least one filter channel interposed 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 permit the at least one liquid to flow therethrough, the at least one filter channel being configured to prevent the plurality of particles from flowing therethrough.

107. The fluidic apparatus of claim 106, the at least one filter channel being formed in at least one surface of the elastic layer.

108. The fluidic apparatus of claim 107, the elastic layer including an upper membrane and a lower membrane.

109. The fluidic apparatus of claim 108, the at least one filter channel being defined by an aperture extending through the upper membrane.

110. A fluidic apparatus comprising:(a) a first layer defining a first chamber portion, the first chamber portion being configured to receive pressurized gas;(b) a second layer defining:(i) a second chamber portion positioned under the first chamber portion, the second chamber portion having a sidewall, the second chamber portion being 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 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 thereby drive the at least one liquid out of the second chamber portion via the plurality of sidewall channels; and(d) a filter interposed 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 permit the at least one liquid to flow therethrough, the filter being configured to prevent the plurality of particles from flowing therethrough, 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 defining a first chamber portion, the first chamber portion being configured to receive pressurized gas;(b) a second layer defining:(i) a second chamber portion positioned under 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 interposed 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.

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

113. The method of any of claims 111 through 112, the plurality of particles comprising a plurality of beads.

114. The method of any of claims 111 through 113, the plurality of particles comprising a plurality of purification particles, the act of treating the at least one liquid comprising purifying the at least one liquid with the plurality of purification particles.

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

116. The method of claim 115. the one or more components including at least one of plasmid DNA or enzymes.

117. The method of any of claims 115 through 116, each of the plurality' of purification particles being configured to separate the mRNA from the components of the transcription reaction process by selectively capturing the mRNA via a polyadenylated (poly A) tail using salt and yvater purification steps.

118. The method of any of claims 114 through 117, each of the plurality of purification particles comprising a rigid, polymeric resin support matrix.

119. The method of claim 118, the rigid, polymeric resin support matrix comprising cross-linked poly(styrene-divinylbenzene).

120. The method of any of claims 114 through 1 19, each of the plurality of purification particles having a polyhydroxyl surface coating.

121. The method of any of claims 114 through 120, each of the plurality’ of purification particles having a surface functionalized with poly(dT).

122. The method of any of claims 111 through 121, each of the plurality7of particles having a predetermined particle size.

123. The method of claim 122, the predetermined particle size being about 50 pm.

124. The method of any of claims 111 through 123, each of the plurality of particles being porous.

125. The method of any of claims 111 through 124, further comprising deforming the elastic layer into the second chamber portion to thereby 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, the filter permitting the at least one liquid to flow therethrough during the act of deforming, the filter preventing the plurality of particles from flowing therethrough during the act of deforming.

127. The method of any of claims 111 through 126, the second layer further defining a particle inlet channel in fluid communication with the second chamber portion, the method further comprising delivering the plurality of particles to the second chamber portion via the particle inlet channel.

128. The method of any of claims 111 through 127, the first layer further defining a third chamber portion, the third chamber portion being configured to receive pressurized gas, the second layer further defining:(i) a fourth chamber portion positioned under 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 and fourth chamber portions; the method further comprising communicating the at least one liquid and the plurality of particles between the second and fourth chamber portions via the at least one bridging channel.

129. The method of any of claims 111 through 128, the second layer further defining a particle outlet channel in fluid communication with the second chamber portion, the method further comprising 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 defining a first chamber portion, the first chamber portion being configured to receive pressurized gas;(b) a second layer defining:(i) a second chamber portion positioned under 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 interposed 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.

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

132. The method of any of claims 130 through 131. the plurality of particles comprising a plurality of beads.

133. The method of any of claims 130 through 132, the plurality of particles comprising a plurality of purification particles, the act of treating the at least one liquid comprising purifying the at least one liquid with the plurality of purification particles.

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

135. The method of claim 134, the one or more components including at least one of plasmid DNA or enzymes.

136. The method of any of claims 134 through 135, each of the plurality of purification particles being configured to separate the mRNA from the components of the transcription reaction process by selectively capturing the mRNA via a polyadenylated (poly A) tail using salt and water purification steps.

137. The method of any of claims 133 through 136, each of the plurality of purification particles comprising a rigid, polymeric resin support matrix.

138. The method of claim 137, the rigid, polymeric resin support matrix comprising cross-linked poly(styrene-divinylbenzene).

139. The method of any of claims 133 through 138, each of the plurality of purification particles having a polyhydroxyl surface coating.

140. The method of any of claims 133 through 139, each of the plurality of purification particles having a surface functionalized with poly(dT).

141. The method of any of claims 130 through 140, each of the plurality of particles having a predetermined particle size.

142. The method of claim 141, the predetermined particle size being about50 pm.

143. The method of any of claims 130 through 142, each of the plurality of particles being porous.

144. The method of any of claims 130 through 143, further comprising deforming the elastic layer into the second chamber portion to thereby 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, the at least one filter channel permitting the at least one liquid to flow therethrough during the act of deforming, the at least one filter channel preventing the plurality of particles from flowing therethrough during the act of deforming.

146. The method of any of claims 130 through 145, the second layer further defining a particle inlet channel in fluid communication with the second chamber portion, the method further comprising delivering the plurality of particles to the second chamber portion via the particle inlet channel.

147. The method of any of claims 130 through 146, the first layer further defining a third chamber portion, the third chamber portion being configured to receive pressurized gas, the second layer further defining:(i) a fourth chamber portion positioned under 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 and fourth chamber portions; the method further comprising communicating the at least one liquid and the plurality of particles between the second and fourth chamber portions via the at least one bridging channel.

148. The method of any of claims 130 through 147, the second layer further defining a particle outlet channel in fluid communication with the second chamber portion, the method further comprising 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 defining a first chamber portion, the first chamber portion being configured to receive pressurized gas;(b) a second layer defining:(i) a second chamber portion positioned under the first chamber portion, the second chamber portion having a sidewall, the second chamber portion being 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 sidewall;(c) an elastic layer disposed between the first layer and the second layer; and(d) a filter interposed 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 comprising treating the at least one liquid with the plurality of particles within the second chamber portion.

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

151. The method of any of claims 149 through 150, the plurality of particles comprising a plurality of beads.

152. The method of any of claims 149 through 151. the plurality of particles comprising a plurality of purification particles, the act of treating the at least one liquid comprising purifying the at least one liquid with the plurality of purification particles.

153. The method of claim 152, each of the plurality of purification particles being configured to separate mRNA from components of a transcription reaction process.

154. The method of claim 153, the components including at least one of plasmid DNA or enzymes.

155. The method of any of claims 153 through 154, each of the plurality of purification particles being configured to separate the mRNA from the components of the transcription reaction process by selectively capturing the mRNA via a polyadenylated (poly A) tail using salt and water purification steps.

156. The method of any of claims 152 through 155, each of the plurality of purification particles comprising a rigid, polymeric resin support matrix.

157. The method of claim 156, the rigid, polymeric resin support matrix comprising cross-linked poly(styrene-divinylbenzene).

158. The method of any of claims 152 through 157, each of the plurality of purification particles having a polyhydroxyl surface coating.

159. The method of any of claims 152 through 158, each of the plurality of purification particles having a surface functionalized with poly(dT).

160. The method of any of claims 149 through 159, each of the plurality of particles having a predetermined particle size.

161. The method of claim 160, the predetermined particle size being about50 pm.

162. The method of any of claims 149 through 161, each of the plurality' of particles being porous.

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

164. The method of claim 163, the filter permitting the at least one liquid to flow therethrough during the act of deforming, the filter preventing the plurality of particles from flowing therethrough during the act of deforming.

165. The method of any of claims 149 through 164, the second layer further defining a particle inlet channel in fluid communication with the second chamber portion, the method further comprising delivering the plurality of particles to the second chamber portion via the particle inlet channel.

166. The method of any of claims 149 through 165, the first layer further defining a third chamber portion, the third chamber portion being configured to receive pressurized gas, the second layer further defining:(i) a fourth chamber portion positioned under 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 and fourth chamber portions;the method further comprising communicating the at least one liquid and the plurality of particles between the second and fourth chamber portions via the at least one bridging channel.

167. The method of any of claims 149 through 166, the second layer further defining a particle outlet channel in fluid communication with the second chamber portion, the method further comprising removing the plurality of particles from the second chamber portion via the particle outlet channel.

168. The fluidic apparatus of any of claims 2-25, each filter channel of the plurality of filter channels having at least one cross dimension that is substantially greater than about 50 pm.

169. The fluidic apparatus of claim 168, the at least one cross dimension being substantially less than about 250 pm.

170. The fluidic apparatus of any of claims 168-169, the at least one cross dimension including at least one of a width or a depth.

171. The fluidic apparatus of claim 170. the at least one cross dimension including both the width and the depth.

172. The fluidic apparatus of any of claims 170-171, the depth being greater than or equal to about 100 pm.

173. The fluidic apparatus of any of claims 170-172, the width ranging from about 100 pm to about 400 pm.

174. The fluidic apparatus of claim 173, the width being about 200 pm.

175. The fluidic apparatus of any of claims 1-25 or 168-174, the second layer defining at least one backflush channel in fluid communication with the at least one fluid outlet channel, the at least one backflush channel being configured to direct at least one backflush fluid through the filter and into the second chamber portion.

176. The fluidic apparatus of any of claims 26-29, the second layer defining at least one backflush channel in fluid communication with the plurality of filter channels, the at least one backflush channel being configured to direct at least one backflush fluid through the plurality of filter channels and into the second chamber portion.

177. The method of any of claims 31 -49, the second layer further defining at least one backflush channel in fluid communication with the at least one fluid outlet channel, the method further comprising directing at least one backflush fluid from the at least one backflush channel through the filter and into the second chamber portion.

178. The method of any of claims 50-68, the second layer defining at least one backflush channel in fluid communication w ith the plurality of filter channels, the method further comprising directing 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 of claims 69-87, the second layer defining at least one backflush channel in fluid communication with the at least one fluid outlet channel, the method further comprising directing at least one backflush fluid from the at least one backflush channel through the filter and into the second chamber portion.

180. The fluidic apparatus of any of claims 89-105. the at least one filter channel having at least one cross dimension that is substantially greater than about 50 pm.

181. The fluidic apparatus of claim 180, the at least one cross dimension being substantially less than about 250 pm.

182. The fluidic apparatus of any of claims 180-181, the at least one cross dimension including at least one of a width or a depth.

183. The fluidic apparatus of claim 182, the at least one cross dimension including both the width and the depth.

184. The fluidic apparatus of any of claims 182-183, the depth being greater than or equal to about 100 pm.

185. The fluidic apparatus of any of claims 182-184, the width ranging from about 100 pm to about 400 pm.

186. The fluidic apparatus of claim 185, the width being about 200 pm.

187. The fluidic apparatus of any of claims 88-105 or 180-186, the second layer defining at least one backflush channel in fluid communication with the at least one fluid outlet channel, the at least one backflush channel being configured to direct at least one backflush fluid through the filter and into the second chamber portion.

188. The fluidic apparatus of any of claims 106-109. the second layer defining at least one backflush channel in fluid communication with the at least one filter channel, the at least one backflush channel being configured to direct at least one backflush fluid through the at least one filter channel and into the second chamber portion.

189. The fluidic apparatus of claim 110, the second layer defining at least one backflush channel in fluid communication with the at least one fluid channel, the atleast one backflush channel being configured to direct at least one backflush fluid through the filter and into the second chamber portion.

190. The method of any of claims 111-129, the second layer defining at least one backflush channel in fluid communication with the at least one fluid channel, the method further comprising directing 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 of claims 130-148, the second layer defining at least one backflush channel in fluid communication with the at least one filter channel, the method further comprising directing 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.

192. The method of any of claims 149-167, the second layer defining at least one backflush channel in fluid communication with the at least one fluid channel, the method further comprising directing 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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