CN117813516A

CN117813516A - System and method for delivering fluid

Info

Publication number: CN117813516A
Application number: CN202280048233.1A
Authority: CN
Inventors: 埃里克·楚; 塔马斯·齐默尔曼; 本杰明·埃尔德里奇; 肯尼思·乔丹; 温西淼
Original assignee: Nutcracker Therapeutics Inc
Current assignee: Nutcracker Therapeutics Inc
Priority date: 2021-06-04
Filing date: 2022-05-27
Publication date: 2024-04-02
Also published as: NL2028535B1

Abstract

A system includes a chip receiving component, a first fluid handling assembly, a second fluid handling assembly, and a fluid communication path. The chip receiving section is for receiving a processing chip having a microfluidic channel. The first fluid handling assembly is for transferring fluid to a microfluidic channel of a processing chip received by the chip receiving member. The second fluid handling assembly includes a sample support feature for supporting a sample container. The second fluid handling assembly further includes a plurality of sampling heads for selectively transferring fluid from a sample container supported by the sample support feature. The fluid communication path includes a plurality of conduits for providing fluid communication between the first fluid handling assembly and the plurality of sampling heads. The first fluid handling assembly is for further transferring fluid from the fluid communication path to a microfluidic channel of a processing chip received by the chip receiving member.

Description

System and method for delivering fluid

Background

The subject matter discussed in this section should not be assumed to be prior art merely because of its mention in this section. Similarly, the problems 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 is merely representative of various approaches that may themselves correspond to implementations of the claimed technology.

Some currently available techniques for manufacturing and formulating polynucleotide therapeutics (e.g., mRNA therapeutics, etc.) may expose the product to contamination and degradation. Some of the available centralized production may be too expensive, too slow, or susceptible to contamination for use in therapeutic formulations that may include multiple polynucleotide species.

Disclosure of Invention

The development of large-scale polynucleotide manufacturing, production of single patient doses, reduction and in some cases even elimination of contaminated contact points, input and process tracking for meeting clinical manufacturing requirements, and use in point-of-care procedures may advance the use of these treatment modalities. Microfluidic instruments and processes may provide advantages to achieve these goals. It may be desirable to facilitate rapid formulation of several samples of the composition, such as for screening purposes or other purposes. Described herein are devices, systems, and methods for facilitating rapid formulation of several samples of a composition by a microfluidic system to overcome the previously existing challenges and achieve the benefits as described herein. Such microfluidic systems can be used to manufacture and formulate products comprising biomolecules, such as therapeutic agents for personalized care.

One implementation relates to a system that includes a chip receiving component, a first fluid handling assembly, a second fluid handling assembly, and a fluid communication path. The chip receiving section is for receiving a processing chip having a microfluidic channel. The first fluid handling assembly is for transferring fluid to a microfluidic channel of a processing chip received by the chip receiving member. The second fluid handling assembly includes a sample support feature for supporting a sample container. The second fluid handling assembly further includes a plurality of sampling heads for selectively transferring fluid from a sample container supported by the sample support feature. The fluid communication path includes a plurality of conduits for providing fluid communication between the first fluid handling assembly and the plurality of sampling heads. The first fluid handling assembly is for further transferring fluid from the fluid communication path to a microfluidic channel of a processing chip received by the chip receiving member.

In some implementations of the system, such as those described in the preceding paragraphs of this disclosure, the system further includes: an instrument having a housing. The chip receiving component and the first fluid handling assembly are positioned within the housing.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, the second fluid handling assembly is positioned within the housing.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, the system further includes: a first controller for driving operation of the first fluid processing assembly.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, the system further includes: a second controller for driving operation of the second fluid handling assembly.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, the second controller communicates with the first controller.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, the first controller is to drive operation of the second fluid handling assembly.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, the chip receiving component includes a mount.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, the first fluid handling assembly includes a reagent storage rack.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, the first fluid handling assembly is for storing one or more fluids.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, the first fluid handling component is for storing one or more reagents.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, the first fluid component is for storing one or more compositions produced by a processing chip received in the chip receiving component.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, the plurality of conduits includes a plurality of flexible tubes.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, the flexible tubes are removably coupled with one or both of the first fluid handling assembly or the second fluid handling assembly.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, the sample support feature is for supporting a plurality of sample trays having a plurality of sample wells.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, the sample support feature includes tray indexing features for indexing the sample trays relative to the sampling heads.

In some implementations of the system, such as any of those implementations described in any of the preceding paragraphs of this disclosure, at least one of the tray indexing features resiliently bears against the sample tray.

In some implementations of the system, such as any of those implementations described in any of the preceding paragraphs of this disclosure, the sample support feature is for supporting a first reagent sample tray for providing the first reagent. The sample support feature also serves to support a composition sample tray for receiving a composition formed using the processing chip received by the chip receiving assembly. The composition is formed using the first reagent.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, the sample support feature is to support: a second reagent sample tray for providing a second reagent, the composition sample tray for receiving a composition formed using the processing chip received by the chip receiving member, the composition formed using the first reagent and the second reagent.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, the sample support feature is to support: a rinse sample tray for providing a rinse fluid. The sample support feature is also for supporting: a waste sample tray for receiving waste generated by a rinsing process including rinsing one or more of the microfluidic channels of the processing chip received by the chip receiving part.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, the second fluid handling assembly further comprises a sample support feature drive assembly for driving the sample support feature along one or more dimensions to position a sample container supported by the sample support feature relative to the sampling heads.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, the sample support feature drive assembly is for driving the sample support feature along two dimensions in a horizontal plane.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, the second fluid handling assembly further includes a head support actuation assembly. The head support actuation assembly is for driving the sampling heads to position fluid-receiving portions of the sampling heads in fluid held by a sample container supported by the sample support feature.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, the head support actuation assembly is to drive the sampling heads vertically to lower and raise the sampling heads relative to the sample support feature.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, each sampling head includes a body defining a first passageway. Each sampling head further comprises: a hollow shaft disposed within the first passageway of the body. The hollow shaft is for transferring fluid from a sample container supported by the sample support feature to the fluid communication path.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, the first passageway has an inner diameter. The hollow shaft has an outer diameter. The outer diameter is smaller than the inner diameter such that a gap is defined between an outer surface of the hollow shaft and an inner surface of the first passageway.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, the body further defines a lower opening in fluid communication with the gap. The body communicates pressurized air to the lower opening via the gap.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, each sampling head further includes a pneumatic fitting and a second passageway. The second passageway and the pneumatic fitting are in fluid communication with the gap. The pneumatic fitting and the second passage are for delivering pressurized air to the gap.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, each sampling head is configured to drive fluid from a sample container supported by the sample support feature by delivering pressurized air to an interior region of the sample container.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, each sampling head further comprises a sealing member for engaging a portion of the sample container supported by the sample support feature.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, the sealing member comprises an annular gasket.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, the system further includes a biasing member for resiliently urging the sealing member into engagement with the portion of the sample container supported by the sample support feature.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, the second fluid handling assembly further comprises a sample container engagement assembly for selectively engaging a sample container supported by the sample support feature.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, the sample container engagement assembly includes a base and one or more actuators for selectively driving the base into and out of engagement with a sample container supported by the sample support feature.

In some implementations of the system, such as any of those described in any of the preceding paragraphs of this disclosure, the sample support feature comprises a platform.

Another implementation relates to a device that includes a sample support feature for supporting a sample container, a sampling head assembly, and a head support actuation assembly. The sampling head assembly includes a mounting body and a plurality of sampling heads supported by the mounting body. Each sampling head includes a sampling head body and a hollow shaft supported by the sampling head body. The hollow shaft includes a lower end for receiving fluid from a sample container supported by the sample support feature. Each sampling head further includes a sealing member for sealing against a surface of a sample container supported by the sample support feature. Each sampling head further comprises: an opening for delivering pressurized air into a space defined above a volume of fluid in a sample container supported by the sample support feature, thereby driving the fluid from the sample container into the hollow shaft. The head support actuation assembly includes a head support plate and one or more actuators. The mounting body is mounted to the head support plate. The one or more actuators are for driving the head support plate toward the sample support feature to selectively urge the lower end of the hollow shaft into a sample container supported by the sample support feature.

In some implementations of the apparatus, such as those described in the preceding paragraphs of this disclosure, the apparatus further comprises: a plurality of fluid conduits for coupling the sampling head assembly with a fluid handling assembly for delivering fluid from a sample container supported by the sample support feature to microfluidic channels in a processing chip via the fluid handling assembly.

In some implementations of the apparatus, such as any of those described in any of the preceding paragraphs of this disclosure, the apparatus further comprises a sample support feature drive assembly for driving the sample support feature along one or more dimensions to position a sample container supported by the sample support feature relative to the sampling head assembly.

In some implementations of the apparatus, such as any of those described in any of the preceding paragraphs of this disclosure, the sample support feature drive assembly is for driving the sample support feature along two dimensions in a horizontal plane.

In some implementations of the device, such as any of those described in any of the preceding paragraphs of this disclosure, the sampling head body defines a first passageway, the hollow shaft being disposed in the first passageway of the sampling head body.

In some implementations of the device, such as any of those described in any of the preceding paragraphs of this disclosure, the first passageway has an inner diameter. The hollow shaft has an outer diameter. The outer diameter is smaller than the inner diameter such that a gap is defined between an outer surface of the hollow shaft and an inner surface of the first passageway.

In some implementations of the device, such as any of those described in any of the preceding paragraphs of this disclosure, the opening of the sampling head assembly is in fluid communication with the gap. The sampling head body is for delivering pressurized air to the opening via the gap.

In some implementations of the apparatus, such as any of those described in any of the preceding paragraphs of this disclosure, each sampling head further includes a pneumatic fitting and a second passageway. The second passageway and the pneumatic fitting are in fluid communication with the gap. The pneumatic fitting and the second passage communicate pressurized air to the gap.

In some implementations of the device, such as any of those described in any of the preceding paragraphs of this disclosure, the sealing member comprises an annular gasket.

In some implementations of the device, such as any of those described in any of the preceding paragraphs of this disclosure, each sampling head assembly further comprises a resilient member for resiliently urging the seal into engagement with the surface of a sample container supported by the sample support feature.

In some implementations of the device, such as any of those described in any of the preceding paragraphs of this disclosure, the resilient member is interposed between a portion of the mounting body and the sampling head body.

In some implementations of the device, such as any of those described in any of the preceding paragraphs of this disclosure, the resilient member is adapted to compress to accommodate a vertical range of motion of the sampling head body relative to the mounting body.

In some implementations of the apparatus, such as any of those described in any of the preceding paragraphs of this disclosure, the apparatus further comprises: a sample container engagement assembly for selectively engaging a sample container supported by the sample support feature.

In some implementations of the device, such as any of those described in any of the preceding paragraphs of this disclosure, the sample container engagement assembly includes a base and one or more actuators for selectively driving the base into and out of engagement with the sample container supported by the sample support feature.

In some implementations of the apparatus, such as any of those described in any of the preceding paragraphs of this disclosure, the one or more actuators are fixed to the head support plate.

In some implementations of the device, such as any of those described in any of the preceding paragraphs of this disclosure, the base defines a plurality of openings. Each opening is configured to receive a corresponding hollow shaft of the plurality of sampling heads.

Another implementation relates to a device that includes a sample support feature for supporting a sample container, a sampling head assembly, a head support actuation assembly, and a sample container engagement assembly. The sampling head assembly includes a mounting body and a plurality of sampling heads supported by the mounting body. Each sampling head includes a sampling head body and a hollow shaft supported by the sampling head body. The hollow shaft includes a lower end for receiving fluid from a sample container supported by the sample support feature. The head support actuation assembly includes a head support plate and one or more actuators. The mounting body is mounted to the head support plate. The one or more actuators are for driving the head support plate toward the sample support feature to selectively urge the lower end of the hollow shaft into a sample container supported by the sample support feature. The sample container engagement assembly is for selectively engaging a sample container supported by the sample support feature. The sample container engagement assembly includes a base and one or more actuators for selectively driving the base into and out of engagement with a sample container supported by the sample support feature.

In some implementations of the apparatus, such as those described in the preceding paragraphs of this disclosure, the one or more actuators are fixed to the head support plate.

In some implementations of the apparatus, such as any of those described in any of the preceding paragraphs of this disclosure, the apparatus further comprises: a plurality of fluid conduits for coupling the sampling head assembly with a fluid handling assembly for delivering fluid from a sample container supported by the sample support feature to microfluidic channels in a processing chip via the fluid handling assembly.

In some implementations of the apparatus, such as any of those described in any of the preceding paragraphs of this disclosure, the sampling head body defines a first passageway. The hollow shaft is disposed in the first passageway of the sampling head body.

In some implementations of the device, such as any of those described in any of the preceding paragraphs of this disclosure, each sampling head further includes a sealing member and an opening. The sealing member is for sealing against a surface of a sample container supported by the sample support feature. The opening is for delivering pressurized air into a space defined above a volume of fluid in a sample container supported by the sample support feature, thereby driving the fluid from the sample container into the hollow shaft.

Another embodiment relates to a method comprising: a plurality of sampling heads are positioned over a plurality of fluid containers. The method further comprises the steps of: the hollow shafts of the sampling heads are inserted into the plurality of fluid containers. The method further comprises the steps of: the first reagent is driven from the first subset of the fluid containers via the first subset of the hollow shafts toward the microfluidic channels in the processing chip. The method further comprises the steps of: a second reagent is driven toward the microfluidic channel in the processing chip. The method further comprises the steps of: combining the first reagent and the second reagent via the processing chip to form a composition. The method further comprises the steps of: the composition is driven from the processing chip to a second subset of the fluid receptacles via a second subset of the hollow shafts.

In some implementations of the method, such as those described in the preceding paragraphs of the disclosure, the second reagent is contained in a third subset of the fluid containers. The driving the second reagent toward the microfluidic channel in the processing chip includes: the second reagent is driven from the third subset of the fluid containers via the third subset of the hollow shafts.

In some implementations of the method, such as any of those implementations described in any of the preceding paragraphs of this disclosure, the method further includes: the buffer is driven toward the microfluidic channel in the processing chip. The combining the first reagent and the second reagent via the processing chip to form a composition includes: combining the buffer with the first reagent.

In some implementations of the method, such as any of those described in any of the preceding paragraphs of this disclosure, the first agent comprises mRNA.

In some implementations of the method, such as any of those implementations described in any of the preceding paragraphs of this disclosure, the second agent comprises a delivery vehicle.

In some implementations of the method, such as any of those described in any of the preceding paragraphs of this disclosure, the composition comprises an encapsulated mRNA.

In some implementations of the method, such as any of those described in any of the preceding paragraphs of this disclosure, the encapsulated mRNA comprises mRNA encapsulated in a delivery vehicle molecule having nanoparticle geometry.

In some implementations of the method, such as any of those implementations described in any of the preceding paragraphs of this disclosure, the method further includes: the microfluidic channels in the processing chip are perfused.

In some implementations of the method, such as any of those described in any of the preceding paragraphs of this disclosure, the priming includes: target areas in the microfluidic channels in the processing chip are monitored. The perfusion further comprises: the presence of fluid in these target areas is detected. The perfusion further comprises: movement of fluid in the microfluidic channels is prevented in response to detecting the presence of fluid in the target regions.

In some implementations of the method, such as any of those implementations described in any of the preceding paragraphs of this disclosure, the method further includes: the hollow shafts of the sampling heads are removed from the plurality of fluid containers.

In some implementations of the method, such as any of those implementations described in any of the preceding paragraphs of this disclosure, the method further includes: the plurality of fluid containers are coupled to the base prior to inserting the hollow shafts of the sampling heads into the plurality of fluid containers. The method further comprises the steps of: after removing the hollow shafts of the sampling heads from the plurality of fluid containers, the plurality of fluid containers are released from the base.

In some implementations of the method, such as any of those implementations described in any of the preceding paragraphs of this disclosure, the method further includes: after driving the composition from the processing chip to the second subset of the fluid containers via the second subset of the hollow shafts, the microfluidic channels in the hollow shafts and the processing chip are rinsed.

In some implementations of the method, such as any of those implementations described in any of the preceding paragraphs of this disclosure, the method further includes: after rinsing the hollow shafts and the microfluidic channels in the processing chip, the hollow shafts and the microfluidic channels in the processing chip are dried.

In some implementations of the method, such as any of those implementations described in any of the preceding paragraphs of this disclosure, the method further includes: the flushing waste fluid in a third subset of these fluid containers is collected. The rinse waste fluid is generated by rinsing the hollow shafts and the microfluidic channels in the processing chip.

In some implementations of the method, such as any of those implementations described in any of the preceding paragraphs of this disclosure, the method further includes: it is determined whether another subset of the fluid containers contains more of the first reagent.

In some implementations of the method, such as any of those implementations described in any of the preceding paragraphs of this disclosure, the method further includes: it is determined that the third subset of the fluid containers does not contain more of the first reagent. The method further comprises the steps of: a plurality of sampling heads are positioned over the third subset of fluid containers. The method further comprises the steps of: the hollow shafts of the sampling heads are inserted into the third subset of fluid containers. The method further comprises the steps of: the first reagent is driven from the third subset of the fluid containers via the first subset of the hollow shafts toward the microfluidic channels in the processing chip. The method further comprises the steps of: a second reagent is driven toward the microfluidic channel in the processing chip. The method further comprises the steps of: combining the first reagent and the second reagent via the processing chip to form a subsequent composition. The method further comprises the steps of: the subsequent composition is driven from the processing chip to a fourth subset of the fluid containers via the second subset of the hollow shafts.

In some implementations of the method, such as any of those implementations described in any of the preceding paragraphs of this disclosure, the method further includes: it is determined that another subset of the fluid containers does not contain more of the first reagent.

The method further comprises the steps of: alerting the user to the completion of the composition forming process.

Another implementation relates to a processor-readable medium comprising content for causing a processor to process data by performing a method such as any of those described in any of the preceding paragraphs of the present disclosure.

It is to be understood that all combinations of the foregoing concepts and additional concepts discussed in more detail below (assuming such concepts are not mutually inconsistent) are considered to be part of the inventive subject matter disclosed herein and to achieve the benefits described herein.

Drawings

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:

FIG. 1 shows a schematic diagram of an example of a system including a microfluidic processing chip;

FIG. 2 shows an exploded perspective view of an example of components of the system of FIG. 1;

FIG. 3 illustrates a top plan view of an example of a processing chip that may be incorporated into the system of FIG. 1;

FIG. 4 schematically illustrates an example of a method of making an mRNA therapeutic composition;

FIG. 5 shows a top plan view of an example of a mixing stage that may be incorporated into a processing chip for formulating mRNA with a delivery vehicle;

FIG. 6 shows a schematic diagram of an example of a system including an instrument for processing fluids on a processing chip and an additional fluid processing subsystem;

FIG. 7 shows a schematic diagram of an example of a system including an instrument having a first fluid handling assembly and a second fluid handling assembly;

FIG. 8 shows a schematic diagram of an example of a system that may be used to prepare several samples of a composition;

FIG. 9 illustrates a perspective view of an example of a fluid handling assembly;

FIG. 10A illustrates a top plan view of the fluid handling assembly of FIG. 9 with the tray support platform in a first position;

FIG. 10B illustrates a top plan view of the fluid handling assembly of FIG. 9 with the tray support platform in a second position;

FIG. 10C illustrates a top plan view of the fluid handling assembly of FIG. 9 with the tray support platform in a third position;

FIG. 11A illustrates a side elevational view of the fluid handling assembly of FIG. 9 with the head support plate in a first position;

FIG. 11B illustrates a side elevational view of the fluid handling assembly of FIG. 9 with the head support plate in a second position;

FIG. 12 shows a perspective view of a tray support platform of the fluid handling assembly of FIG. 9 loaded with a plurality of sample trays;

FIG. 13A shows a top plan view of a portion of the tray support platform of FIG. 12 prior to a sample tray being loaded onto the tray support platform;

FIG. 13B shows a top plan view of a portion of the tray support platform of FIG. 13A with the sample tray on the tray support platform in a non-indexed position;

FIG. 13C shows a top plan view of a portion of the tray support platform of FIG. 13A with the sample tray on the tray support platform in an indexed position;

FIG. 14 illustrates a side elevational view of a portion of the tray support platform of FIG. 12 with alignment features maintaining the sample tray in an indexed position;

FIG. 15 illustrates a perspective view of an example of a sampling head assembly of the fluid handling assembly of FIG. 9, with the sampling head detached from a body of the sampling head assembly;

FIG. 16 illustrates an exploded perspective view of the sampling head assembly of FIG. 15;

FIG. 17 shows a cross-sectional side view of the body of the sampling head of FIG. 16;

FIG. 18 shows a cross-sectional side view of a lower portion of the sampling head of FIG. 16;

FIG. 19 shows a cross-sectional side view of an upper portion of the sampling head of FIG. 16;

FIG. 20 illustrates a perspective view of a tray engagement assembly of the fluid handling assembly of FIG. 9;

FIG. 21 illustrates another perspective view of the tray engagement assembly of FIG. 20;

FIG. 22A illustrates a side elevational view of the head support plate, sampling head assembly and tray engagement assembly of the fluid handling assembly of FIG. 9 in a first operational state relative to a sample tray;

FIG. 22B shows a side elevation view of the components of FIG. 22A in a second operational state relative to the sample tray;

FIG. 22C shows a side elevational view of the component of FIG. 22A in a third operational state with respect to the sample tray;

FIG. 22D shows a side elevation view of the component of FIG. 22A in a fourth operational state with respect to the sample tray;

FIG. 22E shows a side elevation view of the components of FIG. 22A in a fifth operational state relative to the sample tray;

FIG. 22F shows a side elevational view of the component of FIG. 22A in a sixth operational state with respect to the sample tray;

FIG. 22G illustrates a side elevational view of the component of FIG. 22A in a seventh operational state with respect to the sample tray; and is also provided with

FIG. 23 illustrates a flow chart of an example of a method that may be performed using the fluid handling assembly of FIG. 9.

Detailed Description

In some aspects, disclosed herein are devices and methods for treating therapeutic polynucleotides. In particular, the devices and methods may be closed path devices and methods configured to minimize or eliminate manual manipulation during operation. The closed path devices and methods may provide an almost completely sterile environment, and the components may provide a sterile path for processing from an initial input (e.g., template) to an output (e.g., composite therapeutic agent). The materials (e.g., nucleotides and any chemical components) input into the device may be sterile; and can be entered into the system with little or no manual interaction required.

The devices and methods described herein can be used to generate therapeutic agents with rapid cycle times and a high degree of reproducibility. The devices described herein may be configured to provide synthesis, purification, dialysis, compounding, and concentration of one or more therapeutic compositions in a single integrated device. Alternatively, one or more of these processes may be performed in two or more devices as described herein. In some scenarios, the therapeutic composition may include a therapeutic polynucleotide, such as, for example, ribonucleic acid or deoxyribonucleic acid. Polynucleotides may include only natural nucleotide units or may include any kind of synthetic, semisynthetic or modified nucleotide units. All or some of the processing steps may be performed in an uninterrupted fluid processing path that may be configured as one or a series of consumable microfluidic path devices, also referred to herein as a processing chip or biochip in some cases (although the chip need not necessarily be used for bio-related applications). In some examples, the processing chip may be removably mounted in an instrument that is part of a larger microfluidic system, such as shown in fig. 1.

The disclosed devices and methods can be used to synthesize patient-specific therapeutics, including compounding, at the point of care (e.g., hospital, clinic, pharmacy, etc.).

I. Terminology

Throughout the specification and the claims which follow, unless the context requires otherwise, the word "comprise", and variations such as "comprises" and "comprising", will be understood to mean that various elements may be used in both methods and articles of manufacture (e.g. compositions and apparatus including devices 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 devices and methods described herein should be understood to be inclusive, but all or a subset of the elements and/or steps may alternatively be exclusive, and may be expressed as "consisting of, or alternatively" consisting essentially of, the various elements, steps, sub-elements, or sub-steps.

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 "/".

Spatially relative terms, such as "below," "beneath," "lower," "above," "upper," and the like, may be used herein for ease of description to describe one element or feature's relationship to another element or feature as illustrated. It will be understood that 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 the device in the figures is turned over, elements described as "below" or "beneath" other elements or features would then be oriented "above" the other elements or features. Thus, the term "below" can include both above and below orientations. 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 "upward," "downward," "vertical," "horizontal," and the like are used herein for purposes of explanation only, unless explicitly indicated otherwise.

When a feature or element is referred to herein as being "on" another feature or element, it can 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 can 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 illustrated with respect to one embodiment, the features and elements so described or illustrated may be applied to other embodiments. Those skilled in the art will also appreciate that a reference to a structure or feature that is disposed "adjacent" another feature may have portions that overlap or underlie the adjacent feature.

As used herein in the specification and claims, including as used in the examples and unless otherwise explicitly stated, all numbers may be read as if prefaced by the word "about" or "approximately", even if the term does not expressly appear. When describing magnitude and/or position, the phrase "about" or "approximately" may be used to indicate that the value and/or position being described is within a reasonably expected range of values and/or positions. For example, a numerical value may have a value of ±0.1% of the value (or range of values), ±1% of the value (or range of values), ±2% of the value (or range of values), ±5% of the value (or range of values), ±10% of the value (or range of values), and the like. Any numerical values set forth herein should also be understood to include about or approximate such values 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.

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

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 otherwise indicated by the context. These terms are used to distinguish one feature/element from another feature/element and do not denote a particular order unless otherwise indicated. 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.

As used herein, the terms "system," "apparatus," and "device" are to be understood as being interchangeable with one another. The systems, devices, and apparatus may each include multiple components in various structural and/or functional relationships with one another.

As used herein, "polynucleotide" refers to a nucleic acid molecule comprising multiple nucleotides, and generally refers to "oligonucleotides" (polynucleotide molecules of 18-25 nucleotides in length) and polynucleotides of 26 or more nucleotides. Aspects of the disclosure include compositions comprising: oligonucleotides of 18-25 nucleotides in length (e.g., 18-mer, 19-mer, 20-mer, 21-mer, 22-mer, 23-mer, 24-mer, or 25-mer), or medium length polynucleotides of 26 or more nucleotides in length (e.g., 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 200, about 220, about 210, about 250, about 210, about 250, or about 250 nucleotides), or long polynucleotides greater than about 300 nucleotides in length (e.g., between about 300 and about 400 nucleotides in length, between about 400 and about 500 nucleotides in length, between about 500 and about 600 nucleotides in length, between about 600 and about 700 nucleotides in length, between about 700 and about 800 nucleotides in length, between about 800 and about 900 nucleotides in length, between about 900 and about 1000 nucleotides in length, between about 300 and about 500 nucleotides in length, between about 300 and about 600 nucleotides in length, between about 300 and about 700 nucleotides in length, between about 300 and about 800 nucleotides in length, between about, between about 300 and about 900 nucleotides, or about 1000 nucleotides, or polynucleotides even greater than about 1000 nucleotides in length). Where the polynucleotide is double stranded, its length can similarly be described in terms of base pairs.

As used herein, "amplification" may refer to polynucleotide amplification. Amplification may include any suitable method for amplifying 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.

As used herein, a "cassette" (e.g., a synthetic in vitro transcription promoter cassette) refers to a polynucleotide sequence that may include or be operably linked to one or more expression elements, such as an enhancer, promoter, leader sequence, intron, 5 'untranslated region (UTR), 3' UTR, or transcription termination sequence. In some aspects, the 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. As described below, the template may comprise a sequence of interest, such as an open reading frame of interest ("ORF"). The cartridge may be provided as a single element or as two or more unconnected elements.

As used herein, "template" refers to a nucleic acid sequence comprising a sequence of interest for preparing a therapeutic polynucleotide according to the methods of the present disclosure. The template may be, but is not limited to, double-stranded DNA (dsDNA), an engineered plasmid construct, a cDNA sequence, or a linear nucleic acid sequence (e.g., a linear template generated by PCR or by annealing chemically synthesized oligonucleotides). In certain aspects, templates may be integrated into a "box" as described above.

As used herein, the term "sequence of interest" refers to a polynucleotide sequence whose use may be deemed desirable for suitable purposes, particularly for the manufacture of mRNA for therapeutic use, and includes, but is not limited to, coding sequences for structural genes and non-coding regulatory sequences that are not encoded, as well as mRNA or protein products.

As used herein, "in vitro transcription" or "IVT" refers to a process in which in vitro transcription occurs in a non-cellular system to produce synthetic RNA molecules (e.g., synthetic mRNA) for a variety of applications including therapeutic delivery to a subject, e.g., as a therapeutic polynucleotide, which may be part of a therapeutic polynucleotide composition as described below, or may be used to form a therapeutic polynucleotide composition as described below. The resulting therapeutic polynucleotide (e.g., a synthetic RNA molecule (transcript)) can be combined with a delivery vehicle to form a therapeutic polynucleotide composition. Synthetic transcripts include mRNA, antisense RNA molecules, shRNA, circular RNA molecules, ribozymes, and the like. The IVT reaction may use a purified linear DNA template comprising the sequence of the Open Reading Frame (ORF) of the promoter sequence and the sequence of interest, ribonucleotide triphosphates or modified ribonucleotide triphosphates, a buffer system comprising DTT and magnesium ions, and phage RNA polymerase.

As used herein, a "therapeutic polynucleotide" refers to a polynucleotide (e.g., mRNA) that may be part of a therapeutic polynucleotide composition for delivery to a subject to treat a symptom, disease, or disorder in the subject; preventing a symptom, disease, or condition in a subject; improving or otherwise moderating the health of the subject.

As used herein, a "therapeutic polynucleotide composition" (or simply "therapeutic composition") may refer to a composition comprising one or more therapeutic polynucleotides (e.g., mRNA) encapsulated by a delivery vehicle, which composition may be administered to a subject in need thereof using any suitable route of administration, such as intratumoral injection, intramuscular injection, and the like. An example of a therapeutic polynucleotide composition is an mRNA (therapeutic) nanoparticle comprising at least one mRNA encapsulated by a delivery carrier molecule. mRNA vaccines are one example of therapeutic polynucleotide compositions.

As used herein, a "delivery vehicle" refers to any substance that at least partially facilitates in vivo, in vitro, or ex vivo delivery of a polynucleotide (e.g., a therapeutic polynucleotide) to a target cell or tissue (e.g., a tumor, etc.). The designation of something as a delivery vehicle does not necessarily exclude the possibility that the delivery vehicle also has a therapeutic effect. Some versions of the delivery vehicle may provide additional therapeutic effects. In some versions, the delivery vehicle may be a peptoid molecule, such as an aminolipidated peptoid molecule, useful for at least partially encapsulating mRNA. The term "DV" will also be used herein as shorthand for "delivery vehicle".

As used herein, "binding" refers to a method for coupling one component to another component, such as conjugation, synthesis, primer extension, annealing, recombination, or hybridization.

As used herein, "purifying" refers to physically and/or chemically separating one component (e.g., particles) from other unwanted components (e.g., contaminants, fragments, etc.).

As used herein, the term "substantially free" as used with respect to a given substance includes 100% free of the given substance, or it contains less than about 1.0%, or less than about 0.5%, or less than about 0.1% of the given substance.

Overview of a System comprising a microfluidic processing chip

FIG. 1 illustrates an example of various components that may be incorporated into a system (100). The system (100) of this example includes a housing (103) for enclosing a mount (115) capable of removably holding one or more microfluidic processing chips (111). In other words, the system (100) comprises a chip receiving component configured to removably house the processing chip (111), wherein the processing chip (111) itself defines one or more microfluidic channels or fluid paths. The components of the system (100) that fluidly interact with the processing chip (111) (e.g., within the housing (103)) may include fluidic channels or paths that are not necessarily considered to be microfluidics (e.g., such fluidic channels or paths are larger than the microfluidic channels or fluid paths in the processing chip (111)). In some versions, the processing chip (111) is provided and used as a disposable device, while the remainder of the system (100) is reusable. The housing (103) may be in the form of a chamber, a shell, or the like having an opening that may be closed (e.g., via a lid or door, etc.) to thereby seal the interior. The housing (103) may enclose the thermal regulator and/or may be configured to enclose in a thermally conditioned environment (e.g., a refrigeration unit, etc.). The housing (103) may form a sterile barrier. In some variations, the housing (103) may form a humidity or humidity controlled environment. Additionally or alternatively, the system (100) may be located in a cabinet (not shown). Such cabinets may provide a temperature regulated (e.g., refrigerated) environment. Such cabinets may also provide air filtration and air flow management, and may facilitate maintaining the reagents at a desired temperature throughout the manufacturing process. Furthermore, such cabinets may be equipped with UV lamps for sterilizing the processing chips (111) and other components of the system (100). Other suitable features may be incorporated into the cabinet housing the system (100).

In some scenarios, an assembly formed by the housing (103) and components of the system (100) within the housing (103) other than the processing chip (111) may be considered an "instrument". Although the controller (121) and the user interface (123) are shown in fig. 1 as being external to the housing (103), the controller (121) and the user interface (123) may in fact be disposed in or on the housing (103) and thus may also form part of the instrument. As described in detail below, the instrument may removably receive a processing chip (111) via a seat mount (115). When the processing chip (111) is positioned in the mount (115), the instrument and the processing chip (111) cooperate to form together the system (100). When the processing chip (111) is removed from the mount (115), the remaining portion of the system (100) may be considered an "instrument". The instrument, system (100), and processing chip (111) may each be considered a "device". The term "apparatus" may thus be read to include the instrument itself, the processing chip (111) itself, a combination of the instrument and the processing chip (111), some other combination of components of the system (100), or some other arrangement of components of the system (100) or its components.

The mount (115) may be configured to secure the processing chip (111) using one or more pins or other components configured to hold the processing chip (111) in a fixed and predetermined orientation. The mount (115) may thus facilitate maintaining the processing chip (111) in a proper position and orientation relative to other components of the system (100). In this example, the mount (115) is configured to hold the processing chip (111) in a horizontal orientation such that the processing chip (111) is parallel to the ground.

In some variations, a thermal control (113) may be positioned adjacent to the mount (115) to regulate the temperature of any processing chip (111) mounted in the mount (115). The thermal control (113) may include a thermoelectric component (e.g., a peltier device, etc.) and/or one or more heat sinks for controlling the temperature of all or a portion of any of the processing chips (111) mounted in the mount (115). In some variations, more than one thermal control (113) may be included, such as to individually adjust the temperature of different ones of one or more regions of the processing chip (111). The thermal controls (113) may include one or more thermal sensors (e.g., thermocouples, etc.) that may be used to process feedback control of the chip (111) and/or the thermal controls (113).

As shown in fig. 1, the fluid interface assembly (109) couples the processing chip (111) with the pressure source (117) to provide one or more paths for fluid (e.g., gas) at positive or negative pressure to pass from the pressure source (117) to one or more interior regions of the processing chip (111), as will be described in detail below. Although only one pressure source (117) is shown, the system (100) may include two or more pressure sources (117). In some scenarios, pressure may be generated by one or more sources other than the pressure source (117). For example, one or more vials or other fluid sources within the reagent storage rack (107) may be pressurized. Additionally or alternatively, reactions and/or other processes performed on the processing chip (111) may generate additional fluid pressure. In this example, the fluid interface assembly (109) also couples the processing chip (111) with the reagent storage rack (107) to provide one or more paths for liquid reagents or the like to pass from the reagent storage rack (107) to one or more interior regions of the processing chip (111), as will be described in detail below.

In some versions, pressurized fluid (e.g., gas) from at least one pressure source (117) reaches the fluid interface assembly (109) via the reagent storage rack (107) such that the reagent storage rack (107) includes one or more components interposed in a fluid path between the pressure source (117) and the fluid interface assembly (109). In some versions, one or more pressure sources (117) are directly coupled with the fluid interface assembly such that positive pressure fluid (e.g., positive pressure gas) or negative pressure fluid (e.g., suction or other negative pressure gas) bypasses the reagent storage rack (107) to the fluid interface assembly (109). Regardless of whether the fluid interface assembly (109) is interposed in the fluid path between the pressure source (117) and the fluid interface assembly (109), the fluid interface assembly (109) can be removably coupled to the remainder of the system (100) such that at least a portion of the fluid interface assembly (109) can be removed for sterilization between uses. As described in detail below, the pressure source (117) may selectively pressurize one or more chamber areas on the processing chip (111). Additionally or alternatively, the pressure source may also selectively pressurize one or more vials or other fluid storage containers held by the reagent storage rack (107).

The reagent storage rack (107) is configured to contain a plurality of fluid sample holders, each of which may hold a fluid vial configured to hold reagents (e.g., nucleotides, solvents, water, etc.) for delivery to the processing chip (111). In some versions, one or more fluid vials or other storage containers in the reagent storage rack (107) may be configured to receive products from the interior of the processing chip (111). Additionally or alternatively, the second processing chip (111) may receive the product from within the first processing chip (111) such that one or more fluids are transferred from one processing chip (111) to another processing chip (111). In some such scenarios, the first processing chip (111) may perform a first dedicated function (e.g., synthesis, etc.), while the second processing chip (111) performs a second dedicated function (e.g., encapsulation, etc.). The reagent storage rack (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 applicable to the processing chip (111). Such pressure lines may be controlled independently or jointly (in sub-combinations).

The fluid interface assembly (109) may include a plurality of fluid lines and/or pressure lines, wherein each such line includes a biased (e.g., spring-loaded) retainer or tip that individually and independently drives each fluid line and/or pressure line to the processing chip (111) when the processing chip (111) is held in the seat mount (115). Any associated tubing (e.g., fluid lines and/or pressure lines) may be part of the fluid interface assembly (109) and/or may be connected to the fluid interface assembly (109). In some versions, each fluid line includes a flexible tubing connected between a reagent storage rack (107) and a processing chip (111), the reagent storage rack coupling the vials to the tubing in locking engagement (e.g., a ferrule) via a connector. In some versions, the ends of the fluid/pressure lines may be configured to seal against the processing chip (111) (e.g., at corresponding sealing ports formed in the processing chip (111)), as described below. In this example, the connection between the pressure source (117) and the processing chip (111) and the connection between the vials in the reagent storage rack (107) and the processing chip (111) both form sealed closed paths, which paths are isolated when the processing chip (111) is placed in the seat mount (115). Such a sealed closed path may provide protection against contamination when treating therapeutic polynucleotides.

The vials of the reagent storage rack (107) may be pressurized (e.g., >1atm pressure, such as 2atm, 3atm, 5atm, or higher). In some versions, the vial may be pressurized by a pressure source (117). Thus either negative or positive pressure may be applied. For example, the fluid vials may be pressurized to about 1psig to about 20psig (e.g., 5psig, 10psig, etc.). Alternatively, a vacuum (e.g., about-7 psig or about 7 psia) may be applied at the end of the process to draw the fluid back into the vial (e.g., the vial used as a reservoir). The fluid bottle may be driven at a low pressure, such as a pneumatic valve described below, which may prevent or reduce leakage. In some variations, the pressure differential between the fluid valve and the pneumatic valve may be between about 1psi and about 25psi (e.g., about 3psi, about 5psi, 7psi, 10psi, 12psi, 15psi, 20psi, etc.).

The system (100) of the present example also includes a magnetic field applicator (119) configured to form a magnetic field at a region of the processing chip (111). The magnetic field applicator (119) may include a movable head operable to move a magnetic field to selectively isolate products of magnetically captured beads adhered to vials or other storage containers in the reagent storage rack (107).

The system (100) of the present example also includes one or more sensors (105). In some versions, such sensors (105) include one or more cameras and/or other types of optical sensors. Such sensors (105) may sense one or more of a bar code, a liquid level within a fluid vial held within a reagent storage rack (107), fluid movement within a processing chip (111) mounted within a mount (115), and/or other optically detectable conditions. In versions where the sensor (105) is used to sense a bar code, such bar code may be included on vials in the reagent storage rack (107) such that the sensor (105) may be used to identify vials in the reagent storage rack (107). In some versions, a single sensor (105) is positioned and configured to simultaneously view such bar codes on vials in the reagent storage rack (107), liquid levels in vials in the reagent storage rack (107), fluid movement within a processing chip (111) mounted within the 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 individually view corresponding optically detectable conditions such that the sensors (105) may be dedicated to a particular corresponding optically detectable condition.

In versions where the sensor (105) includes at least one optical sensor, visual/optical markers may be used to estimate yield. For example, fluorescence can be used to detect process yields or residual materials by labeling with fluorophores. In addition or in the alternative, dynamic Light Scattering (DLS) may be used to measure particle size distribution within a portion of the processing chip (111), such as a hybrid portion of the processing chip (111), for example. In some variations, the sensor (105) may transmit light (e.g., laser light) to the processing chip (111) using one or two optical fibers and detect the light signal emitted from the processing chip (111), thereby providing a measurement result. In versions where the sensor (105) optically detects process yield or residual material, etc., the sensor (105) may be configured to detect visible light, fluorescence, ultraviolet (UV) absorption signals, infrared (IR) absorption signals, and/or any other suitable kind of optical feedback.

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

The system (100) of the present example may be controlled by a controller (121). The controller 121 may include one or more processors, one or more memories, and various other suitable electronic components. In some versions, one or more components of the controller (121) (e.g., one or more processors, etc.) are embedded within the system (100) (e.g., housed within the housing (103)). Additionally or alternatively, one or more components of the controller (121) (e.g., one or more processors, etc.) may be removably attached or removably connected with other components of the system (100). Thus, at least a portion of the controller (121) may be removable. Furthermore, in some versions, at least a portion of the controller (121) may be remote from the housing (103).

Control of the controller (121) may include activating the pressure source (117) to apply pressure through the processing chip (111) to drive fluid movement, among other tasks. The controller (121) may be wholly or partially external to the housing (103); either completely or partially inside the housing (103). The controller (121) may be configured to receive user input via a user interface (123) of the system (100); and provides output to a user via a user interface (123). In some versions, the controller (121) is fully automated, requiring no user input. In some such versions, the user interface (123) may provide output to the user only. The user interface (123) may include a monitor, touch screen, keyboard, and/or any other suitable feature. The controller (121) may coordinate processing including moving one or more fluids onto the processing chip (111), mixing one or more fluids onto the processing chip (111), adding one or more components to the processing chip (111), metering fluids on the processing chip (111), adjusting the temperature of the processing chip (111), applying a magnetic field (e.g., when using magnetic beads), and the like. The controller (121) may receive real-time feedback from the sensor (105) and execute a control algorithm based on such feedback from the sensor (105). Such feedback from the sensor (105) may include, but is not necessarily limited to, identification of the reagent in the vial in the reagent storage rack (107), detected level in the vial in the reagent storage rack (107), detected movement of the fluid on the processing chip (111), fluorescence of the fluorophore in the fluid on the processing chip (111), and the like. The controller (121) may include software, firmware, and/or hardware. The controller (121) may also communicate with a remote server, for example, to track the operation of the device, reorder materials (e.g., components such as nucleotides, processing chips (111), and/or download schemes, etc.).

Fig. 2 illustrates an example of some forms that various components of the system (100) may take. Specifically, fig. 2 shows a reagent storage rack (150), a fluidic interface assembly (152), a seat mount (154), a thermal control (156), and a processing chip (200). The reagent storage rack (150), the fluid interface assembly (152), the seat mount (154), the thermal control (156), and the processing chip (200) of this example may be constructed and operated as the reagent storage rack (107), the fluid interface assembly (109), the seat mount (115), the thermal control (113), and the processing chip (111), respectively, described above. These components are fixed relative to the base (180). A set of rods (182) support the reagent storage rack (150) on the fluid interface assembly (152).

As shown in fig. 2, a set of optical sensors (160) are positioned at four respective locations along the base (180). The optical sensor (160) may be configured and operated as the sensor (105) described above. The optical sensor (160) may comprise an off-the-shelf camera or any other suitable kind of optical sensor. The optical sensors (160) are positioned such that a fluid vial held within the reagent storage rack (150) is within the field of view of one or more of the optical sensors (160). In addition, the processing chip (200) is within a field of view of the one or more optical sensors (160). Each optical sensor (160) is movably secured to the base (180) via a corresponding track (184) (e.g., in a gantry arrangement) such that each optical sensor (160) is configured to translate laterally along each corresponding track (184). A linear actuator (186) is secured to each optical sensor (160) and is thereby operable to drive the lateral translation of each optical sensor (160) along the corresponding track (184). Each actuator (186) may be in the form of a drive belt, a drive chain, a drive cable, or any other suitable type of structure. The controller (121) may drive operation of the actuator (186). The optical sensor (160) may be moved along the track (184) during operation of the system (100) to facilitate viewing of the appropriate area of vials in the reagent storage rack (150) and/or processing the chip (200). In some scenarios, the optical sensor (160) moves in unison along the corresponding track (184). In some other scenarios, the optical sensors (160) move independently along the corresponding tracks (184).

Although the optical sensor (160) is shown mounted to the base (180) in fig. 2, the optical sensor (160) may be positioned elsewhere within the system (100) in addition to or instead of being mounted to the base (180). For example, some versions of the reagent storage rack (107) may include one or more optical sensors (160) positioned and configured to provide a top field of view. In some such versions, such optical sensors (160) may be mounted to rails, movable cantilevers, or other structures that allow such optical sensors (160) to be repositioned during operation of the system (100). The optical sensor (160) may be positioned at any other suitable location. Although not shown, the system (100) may also include one or more light sources (e.g., electroluminescent panels, etc.) to provide illumination that facilitates optical sensing of the optical sensor (160).

In some versions, one or more mirrors are used to facilitate visualization of components of the system (100) by the optical sensor (160). Such mirrors may allow the optical sensor (160) to view components of the system (100) that may not otherwise be within the field of view of the sensor (160). Such a mirror may be arranged directly adjacent to the optical sensor (160). Additionally or alternatively, such mirrors may be disposed adjacent to one or more components of the system (100) to be viewed by the optical sensor (160).

In use of the system (100), an operator may select a solution to be run (e.g., from a library of preset solutions), or a user may input a new solution (or modify an existing solution) via the user interface (123). According to an aspect, the controller (121) may instruct an operator which type of processing chip (111) to use, what the contents of the vials in the reagent storage rack (107) should be, and where to place the vials in the reagent storage rack (107). An operator can load a processing chip (111) into a seat mount (115); and the required reagent vials and output vials are loaded into a reagent storage rack (107). The system (100) can confirm the presence of the desired peripheral devices, identify the processing chip (111), and scan the identifiers (e.g., bar codes) of each reagent and product vial in the reagent storage rack (107), thereby facilitating vial matching of the reagent inventory for the selected protocol. After confirming the starting materials and the equipment, the controller (121) may execute the protocol. During execution, the valve and pump are actuated to deliver the reagents, as described in detail below, the reagents are mixed, the temperature is controlled, and the reaction occurs, a measurement is made, and then the product is pumped to a target vial in a reagent storage rack (107).

Example of a processing chip

Fig. 3 shows an example of the processing chip (200) in more detail. In combination with the remainder of the system (100), the processing chip (200) can be used to provide in vitro synthesis, purification, concentration, formulation, and analysis of therapeutic compositions, including but not limited to therapeutic polynucleotides and therapeutic polynucleotide compositions. As shown in fig. 3, the processing chip (200) of this example includes a plurality of fluid ports (220). Each fluid port (220) has an associated fluid channel (222) formed in the processing chip (200) such that fluid transferred into the fluid port (220) will flow through the corresponding fluid channel (222). As described in detail below, each fluid port (220) is configured to receive fluid from a corresponding fluid line (206) from the fluid interface assembly (109). In this example, each fluid channel (222) leads to a valve chamber (224) operable to selectively block or allow further transfer of fluid from the corresponding fluid channel (222) along the processing chip (200), as will be described in more detail below.

As also shown in fig. 3, the processing 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 a therapeutic composition as described herein. By way of example only, such additional chambers (230, 250, 270) may be used to provide synthesis, purification, dialysis, compounding, and concentration of one or more therapeutic compositions; or perform any other suitable function. Fluid may be transferred from one chamber (230) to another chamber (230) via a fluid connector (232). In some versions, the fluid connector (232) may operate like a valve between an open state and a closed state (e.g., similar to the valve chamber (224)). In some other versions, the fluid connector (232) is left open throughout the preparation of the therapeutic composition. In this example, chamber (230) is used to provide synthesis of polynucleotides, although chamber (230) may alternatively be used for any other suitable purpose.

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

A further valve chamber (252) is interposed between each chamber (250) and the corresponding chamber (270) such that fluid can be selectively transferred from chamber (250) to chamber (270) via valve chamber (252). The chambers (270) are also coupled to each other such that the processing chip (200) can transfer fluid back and forth between the chambers (270). The chamber (270) may be used to provide mixing of fluids and/or may be used for any of the other various purposes described herein; and may have any suitable configuration.

As shown in fig. 3, the chamber (270) is also coupled with the further fluid port (221) via a corresponding fluid channel (223) and valve chamber (225). The fluid port (221), fluid channel (223) and valve chamber (225) may be constructed and operate as the fluid port (220), fluid channel (222) and valve chamber (224) described above. In some versions, the fluid port (221) is used to communicate additional fluid to the chamber (270). Additionally or alternatively, the fluid port (221) may be used to transfer fluid from the processing chip (200) to another device. For example, fluid from the chamber (270) may be transferred directly to another processing chip (200), to one or more vials in the reagent storage rack (107), or elsewhere via the fluid port (221).

The processing chip (200) further comprises a number of memory chambers (260). In this example, each storage chamber (260) is configured to receive and store fluid transferred to or from the corresponding chamber (250, 270). Each storage chamber (260) has a corresponding inlet valve chamber (262) and outlet valve chamber (264). Each inlet valve chamber (262) is interposed between the storage chamber (260) and the corresponding chamber (250, 270) and is thereby operable to permit or prevent fluid flow between the storage chamber (260) and the corresponding chamber (250, 270). Each outlet valve chamber (264) is operable to meter fluid flow between the storage chamber (260) and a corresponding fluid port (266). In some versions, each fluid port (266) is configured to transfer fluid from a corresponding vial in the reagent storage rack (107) to a corresponding storage chamber (260). Additionally or alternatively, each fluid port (266) may be configured to transfer fluid from a corresponding storage chamber (260) to a corresponding vial in the reagent storage rack (107). In this example, the storage chamber (260) is used to provide a metering of fluids transferred to and/or from the processing chip (200). Alternatively, storage chamber (260) may be used for any other suitable purpose, including, but not limited to, pressurizing fluid transferred to and/or from process chip (200).

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

The processing chip (200) may also include electrical contacts, pins, pin receptacles, capacitive coils, inductive coils, or other features configured to provide electrical communication with other components of the system (100). In the example shown in fig. 3, the processing chip (200) includes an electroactive area (212) having such electrical communication features. The electroactive zone (212) may also include electronic circuitry and other electronic components. In some versions, the electroactive zone (212) may provide communication of power, data, and the like. Although the electroactive region (212) is shown at one particular location on the processing chip, the electroactive region (212) may alternatively be positioned at any other suitable location or locations. In some versions, the electroactive region (212) is omitted.

Exemplary methods of manufacturing therapeutic Agents

The above-described systems can be used to manufacture mRNA-based therapeutics or other compositions as described herein. An example of a method for preparing an mRNA therapeutic agent is shown in fig. 4. In this exemplary method, a target sequence ("sequence of interest") is identified, as shown in block (300) of fig. 4. Templates containing the target sequence ("sequence of interest") may then be prepared and amplified ("amplification"), as indicated in block (310). mRNA is produced using a template comprising the target sequence via in vitro transcription of the mRNA as indicated in box (320). The resulting mRNA comprising the sequence of interest may then be purified, as indicated in block 330, and then formulated with DV, as indicated in block 340. The resulting formulation comprising mRNA may then be further processed and optionally purified, as indicated by box (360), for therapeutic use, as indicated by box (360). An example of the details of the method shown in fig. 4 will be described further below.

Therapeutic uses of the compositions produced by the methods shown in fig. 4 may include, for example, cell therapy, tumor therapy, protein replacement, vaccine, expression of effector proteins, loss of function induced by expression of dominant negative proteins, and gene/genome editing. In addition to their high potency, mRNA therapeutics can have benefits associated with their rapid development cycle, standardized manufacturing, transient expression, and low risk of genomic integration. The methods and devices described herein can be used to manufacture mRNA therapeutics for one or more of these therapeutic categories.

A. Identifying sequences of interest

For the portion of the method represented by block (300) of fig. 4, any suitable method and criteria may be used to identify the sequence of interest. In some cases, the sequence of interest may be a short fragment of DNA encoding some or all of the product molecules (RNA or protein). The sequence of interest can be based at least in part on the genetics (e.g., genotype) of the particular patient, including generating a particular mRNA composition based on the sequence of the patient himself. The sequence of interest may additionally or alternatively be based at least in part on the phenotype of the particular patient (e.g., based on the category in which the patient falls, such as a risk factor category). In any event, by the systems and methods described herein, the compositions can be compounded at the point of care to generate an optimized treatment for the individual.

B. Preparation of template (amplification)

Once the sequence of interest has been identified, a template comprising the sequence of interest may be prepared and amplified, as indicated in block (310). The template may be a DNA template, such as linear DNA, plasmid DNA, or a combination thereof. The template may comprise an in vitro transcription promoter cassette (IFC).

The IFCs may be double stranded DNA capable of in vitro transcription. Templates may be incorporated into IFCs having functional elements (e.g., from the inserted sequence of interest) that facilitate in vitro transcription, such as promoters, portions encoding the 5 'untranslated region (5' utr), portions encoding the 3 'untranslated region (3' utr), and portions encoding the polyadenylation tail. The IFCs may also include one or more linkers (e.g., at least one cleavable site) for cloning the sequence of interest into an in vitro transcription promoter cassette for expression of the sequence of interest and for restriction sites to allow linearization of the template. IFCs may be manufactured synthetically or non-synthetically.

Sequences of interest useful for insertion into an IFC may be synthetically or non-synthetically made. The sequence of interest may be cleaved before it is combined with the IFCs. In particular, the sequence of interest may be cleaved with the same restriction endonuclease as that used to cleave the IFCs; but may also be generated by enzymatic amplification. In any event, the templates generated according to block (310) of the method shown in FIG. 4 may take various forms. In some versions, the template includes a uracil-containing polynucleotide sequence.

C. In vitro transcription

A templates generated according to block (310) of the method shown in fig. 4 may be used in a subsequent In Vitro Transcription (IVT) reaction to form therapeutic polynucleotides, such as therapeutic mRNA, as shown in block (320) of fig. 4. The IVT process may be performed inside a processing chip, such as any of the processing chips (111,200) described herein, wherein the processing is driven by a controller (121). Part of the IVT process can include combining the template with reagents such as uracil-N-glycosylase (UNG), dntps (including dUTP, modified dUTP, and combinations thereof), polymerase, and buffers. The IVT reaction is incubated under controlled conditions to produce a capped mRNA molecule. After the IVT reaction, DNAse treatment may be performed to degrade the template DNA. This may be performed inside the IVT reaction chamber and parameters such as dilution rate, enzyme/buffer concentration, temperature and mixing may be controlled to optimal levels. The program may be executed autonomously and recorded by a monitoring camera (e.g., one or more of the sensors (105)).

Purification of mRNA

As indicated at block 330, mRNA generated by the IVT process may be purified to remove impurities and byproducts. In some versions, the purification includes washing with cellulose and ethanol. For example, cellulose membranes can be used to selectively capture dsRNA under precisely controlled binding conditions, and to elute unbound fractions into a chamber of a processing chip, such as processing chip (111,200). Another purification step may use 1 μm-2 μm carboxyl-coated paramagnetic beads that selectively capture mRNA greater than 500bp in length. One or more washes may be performed to remove unbound materials such as nucleotides, enzymes, and degraded templates. The pure mRNA can then be eluted in USP grade water. The sample chamber of the processing chip (111,200) can be used to analyze purified mRNA. The sampling chamber may receive detection reagents/probes to confirm the content of the resulting purified mRNA.

E. mRNA formulation with delivery vehicle

The purified mRNA may then be retrieved from the processing chip (111,200) for formulation with DV, as shown in block (340). In some cases, the formulation process is performed at least in part by processing a formulation version of the chip (111). The purified mRNA can be combined with at least one DV molecule to form mRNA nanoparticles by a formulation process. For example, an aqueous solution of mRNA cargo may be combined with an ethanol solution of DA in a microfluidic mixing structure within a formulated version of the processing chip (111). The material may then undergo two post-formulation treatment steps involving: an on-chip dialysis process is first performed to exchange buffer components in the formulated product, followed by a concentration step to reduce the volume of the drug product to match the specifications. The resulting formulation may produce encapsulated mRNA in the form of Amphiphilic Nanoparticles (ANP). In some versions, the ANPs have a diameter of about 100nm or less.

In some versions, the DV molecule may include a lipid-based molecule, such as an aminolipidated peptoid. During the compounding process of block (340), the temperature of the mixing stage on the compounding version of the processing chip (111) may be controlled to a temperature or temperature range (e.g., between about 2 degrees celsius and about 20 degrees celsius) calibrated to enhance the mixing action for mixing in the mixing stage. The elevated mixing temperature may be based on the formulation (in some examples, sequences of mRNA and/or DV) being mixed within a particular geometry of the mixing chamber. Exposure of DV components to aqueous solutions and interactions between cationic (+) lipids and anionic (-) mRNA can trigger particle formation. mRNA can be dissolved in an acidic buffer, which can help ensure complete protonation of the basic functional group (e.g., amine) on DV responsible for its cationic charge. DV is soluble in aqueous miscible organic solvents (e.g., ethanol) that facilitate the formation of nanoscale particles when exposed to aqueous cargo solutions. Immediately after mixing, the solution pH can be stabilized with neutral buffer.

In some versions, a peptoid-based lipid formulation may be used as a DV that can bind both a cationic group and a lipid moiety to an N-substituted peptide (i.e., peptoid) backbone. The DV component may be a monodisperse, fully characterizable chemical entity. DV may comprise one or more polyanionic compounds, one or more Polyglycolized (PEG) compounds, which refers to covalent binding of polyethylene glycol molecules, and one or more cationic compounds. Suitable cationic compounds may include, but are not limited to, cationic lipids, cationic lipid-peptide conjugates (e.g., lipids), cationic peptides, cationic polymers, and lipid-like (lipophilic) cationic compounds. DV may comprise one or more tertiary amino-lipidated and/or pegylated cationic peptide compounds. The tertiary amino lipidated and/or pegylated cationic peptide compound may be a peptide chain comprising an N-substituted amino acid residue.

The formulation version of the processing chip 111 can precisely control the mixing rate of the materials. Faster or slower mixing may be provided and controlled by the controller (121). In some versions, the ANP may be diluted immediately after mixing by direct addition of 1:1 neutral PBS. This neutralizes acidic formulation buffers and allows for the preparation of formulations for dialysis and concentration. The ANP produced by the configuration process of block (340) may also be evaluated on the formulation version of the processing chip (111). For example, the formulation process of block (340) may include one or more Dynamic Light Scattering (DLS) stages to evaluate the particle size, particle distribution, and/or other characteristics of the ANP. In addition or in the alternative, fluorescent mRNA-specific probes can be used to determine RNA concentration before and after particle disruption by the addition of detergent. This assay can elucidate the mRNA concentration used for dosing information and the percentage of mRNA encapsulated in ANP relative to free mRNA in solution. Other methods may be used.

F. After preparation

Once the ANP is formed during the formulation process of block 340, as shown in block 350 of fig. 4, a number of post-processing operations may be completed on the formulation version of the processing chip 111. In some versions, these additional processes may include dialysis for buffer exchange and ethanol removal, followed by evaporative concentration to reduce the volume for administration. Other suitable processing steps may be used. Finally, the method may produce a ready-to-use therapeutic polynucleotide composition, as indicated in block (360). Such therapeutic compositions may include, but are not limited to, cell therapies, tumor treatments, protein replacement, vaccines, expression of effector proteins, loss of function induced by expression of dominant negative proteins, and gene/genome editing. Such therapeutic compositions may be delivered to a patient in any suitable manner.

The various sub-processes mentioned in fig. 4 may be performed using any suitable number or type of processing chips (111). In some versions, the entire process shown in fig. 4 is performed using a single version of the processing chip (111). In some other versions, some of the sub-processes are performed on a dedicated processing chip (111), while other sub-processes are performed on another dedicated processing chip (111). For example, in some versions, template preparation (block 310) is performed on a dedicated template version of the processing chip (111); IVT transcription and purification (blocks 320, 330) are performed on a dedicated IVT version of the processing chip (111); and, the compounding (block 340) is performed on a dedicated compounding style of the processing chip (111).

Fig. 5 illustrates a portion of a processing chip (400) having features that may be used to perform at least a portion of a compounding process (block 340). The processing chip (400) of this example includes a plurality of fluid channels (402). Each fluid channel (402) has a fluid port (not shown) such that fluid may be transferred to the fluid channel (402) via the corresponding fluid port. Some of these fluid ports may receive fluid from corresponding vials in the reagent storage rack (107). Additionally or alternatively, some of the fluid ports may receive fluid from a corresponding fluid output of another processing chip (111,200). Alternatively, a fluid port to the fluid channel (402) may receive fluid from any other suitable source.

The fluid channels (402) lead to several mixing components (420) integrated into the processing chip (400). In some versions, all mixing components (420) on a processing chip (400) have the same kind of fluid input and are intended to all generate the same kind of fluid output. Each mixing assembly (420) includes a set of vacuum caps (422), a set of inlet valves (424), and a set of mixing chambers (430, 440). Referring to one mixing assembly (420) as representative of the other mixing assemblies (420), the mixing assembly (420) includes a first vacuum cap (422 a) that receives fluid from the first fluid channel (402 a); a second vacuum cap (422 b) that receives fluid from the second fluid channel (402 b); and a third vacuum cap (422 c) that receives fluid from the third fluid channel (402 c). Each vacuum lid (422 a,422b,422 c) is configured to evacuate air or other gas from the corresponding fluid channel (402 a,402b,402 c) such that the vacuum lid (422 a,422b,422 c) may purge any bubbles or the like that may otherwise be present. The first valve (424 a) selectively prevents or allows fluid flow from the first vacuum cap (422 a) into the first inlet channel (426 a) leading to the first mixing chamber (430). The second valve (424 b) selectively prevents or allows fluid flow from the second vacuum cap (422 b) into the inlet channel (426 b) leading to the first mixing chamber (430). The channels (426 a,426 b) converge to form an inlet channel (432) to the first mixing chamber (430). The fluids from the channels (426 a,426 b) are thus mixed together within the first mixing chamber (430).

A third valve (424 c) selectively prevents or allows fluid flow from the third vacuum cap (422 c) into a third channel (426 c) leading to the second mixing chamber (440). The outlet passage (434) from the first mixing chamber (430) converges with the third passage (426 c) to form an inlet passage (442) to the second mixing chamber (440). The fluids from the channels (434, 426 c) are thus mixed together in the second mixing chamber (440). The fluid mixed in the second mixing chamber (440) is output through an outlet channel (444).

In some versions that provide for encapsulation of mRNA using the processing chip (400), the combination of mRNA and formulation buffer may be transmitted through the first fluid channel (402 a), and one or more DV molecules in ethanol may be transmitted through the second fluid channel (402 b). In some versions, the formulation buffer includes an aqueous buffer under slightly acidic conditions (e.g., having a pH of about 6.0), such as a phosphate-citrate buffer. Alternatively, any other suitable formulation buffer may be used. The mRNA and DV molecules may thus be combined for encapsulation in the first mixing chamber (430). A diluent (e.g., phosphate Buffered Saline (PBS) solution) may be delivered through the third fluid channel (402 c). In such versions, the second mixing chamber (440) may thus be used to provide pH adjustment. In some variations, the mRNA and the formulation buffer are combined in another mixing chamber (not shown) upstream of the first fluid channel (402 a). Similarly, the DV molecules and ethanol may be combined in another mixing chamber (not shown) upstream of the second fluid channel (402 b).

The additional channel (452) is fluidly coupled with the outlet channel (444) via an opening (450). The channel (452) may be fluidly coupled to a collection vial (e.g., for storage, etc.) in the reagent storage rack (107), to another processing chip (111,200) (e.g., for further processing, etc.), or to any other object.

In versions where certain sub-processes are performed on a dedicated processing chip (111) and other sub-processes are performed on another dedicated processing chip (111), the same instrument of the system (100) may be used with various processing chips (111). In some such versions, the same instrument of system (100) houses all of the processing chips (111) required to perform the process shown in fig. 4, such that the instrument of system (100) transfers fluid from one processing chip (111) to another processing chip (111) at the appropriate stage of the process. In some other versions, the instrumentation of the system (100) accommodates only a single processing chip (111) at a time. In some such versions, a portion of the process of fig. 4 (e.g., template preparation (block (310)) may be performed using a dedicated processing chip (111), wherein the resulting fluid is stored in one or more vials in the reagent storage rack (107): then, the dedicated processing chip (111) may be removed from the instrument of the system (100) and replaced with another dedicated processing chip (e.g., a version of the processing chip (111) dedicated to performing IVT transcription (block (320)), wherein the second dedicated processing chip receives fluid from one or more vials in the reagent storage rack (107) and/or other sources.

V. examples of automated fluid delivery systems

In some scenarios, it may be desirable to provide additional fluid handling capacity to a system such as system (100). For example, it may be desirable to provide an accessory fluid handling assembly that interfaces with components of the system (100) to allow a user to easily test several samples of reagents during a process performed by the system (100); and several samples of the composition produced by the system (100) are easily retrieved using the reagent samples. In some such scenarios, a user may wash several samples of the DV fluid (e.g., DV molecules in ethanol as described above) in a formulation process performed in a processing chip (e.g., like processing chip (400)) to test several samples of the mRNA fluid (e.g., a combination of mRNA and formulation buffer generated by the IVT and purification process as described above with reference to blocks (320, 330) of fig. 4); and retrieving several samples of the encapsulated mRNA composition (e.g., in the form of ANP) formed during the formulation process.

While experimental tests such as those described above may be performed in the system (100), such as by preloading the reagent storage rack (107) with several reagent samples and retrieving samples of the encapsulated mRNA composition from the reagent storage rack (107), the accessory fluid handling assembly may allow a user to more easily provide a large number of discrete reagent samples and collect a large number of discrete encapsulated mRNA composition samples (e.g., 96 discrete encapsulated mRNA composition samples). In other words, the reagent storage rack (107) itself may have only the ability to "hold a number of reagent samples," which may limit the usability of the reagent storage rack (107) to screen a large number of conditions (e.g., different reagents). For example, some versions of the reagent storage rack (107) may involve a user switching vials in the reagent storage rack (107), washing fluid communication channels to and within the processing chip, and/or performing other potentially time-consuming operations. The accessory fluid handling assembly may provide additional fluid storage and handling capabilities relative to the capabilities of the reagent storage rack (107), thereby increasing the number of conditions that can be screened, automating the use of different reagent samples, and automating the cleaning of fluid channels between reagent samples. The accessory fluid handling assembly may also provide for accurate extraction of reagents, thereby preventing or otherwise reducing waste. Several examples of how the accessory fluid treatment assembly may be combined with or incorporated into a variation of the system (100) will be described in detail below.

A. Examples of arrangements of accessory fluid handling assemblies

Fig. 6 shows an example of a system (500) including an instrument (510) and a separate fluid handling subsystem (520). The example instrument (510) may be configured and operate like the instrument of the system (100). For example, the instrument (510) of this example includes a controller (512) and an integral fluid handling assembly (514). The controller (512) may be configured and operate like the controller (121). The fluid handling assembly (514) may be configured and operated like a combination of the pressure source (117), the reagent storage rack (107), and the fluid interface assembly (109). The controller (512) is coupled with the fluid handling assembly (514) via an electrical communication path (513), which may include a plurality of wires, traces, other conductive paths, wireless links, and the like. The controller (512) is thus operable to drive operation of the fluid processing assembly (514) via the electrical communication path (513).

The apparatus (510) of this example is also operable to removably receive a processing chip (516) that is configurable and operable as any of the variations of the processing chip (111) described herein. The fluid processing assembly (514) may be coupled with the processing chip (516) via a fluid communication path (515), which may include a plurality of tubes, other fluid conduits, and the like. The instrument (510) may also have other components and functions similar to those described above with respect to the instrument of the system (100).

The example fluid handling subsystem (520) includes a controller (522) and a fluid handling assembly (524). The controller (522) may be configured and operate as the other controllers (121,512) described herein. The controller (522) is coupled to the fluid handling assembly (524) via an electrical communication path (523) that may include a plurality of wires, traces, other conductive paths, wireless couplings, and the like. The controller (522) is thus operable to drive operation of the fluid handling assembly (524) via the electrical communication path (523). The controller (522) of the fluid handling subsystem (520) is also coupled with the controller (512) of the instrument (510) via an electrical communication path (530), which may include a plurality of wires, traces, other conductive paths, wireless couplings, and the like. In some versions, controller (522) may communicate commands, data, and/or other signals to controller (512) via electrical communication path (530). Additionally or alternatively, the controller (512) may communicate commands, data, and/or other signals to the controller (522) via the electrical communication path (530). In some variations, the electrical communication path (530) is omitted such that the controllers (512,522) are not in electrical communication with each other.

The fluid handling assembly (524) of the fluid handling subsystem (520) is coupled with the fluid handling assembly (514) of the instrument (510) via a fluid communication path (532), which may include a plurality of tubes, other conduits, and the like. In some versions, the fluid handling assembly (524) may communicate fluid to the fluid handling assembly (514) via a fluid communication path (532). Additionally or alternatively, the fluid handling assembly (514) may communicate fluid to the fluid handling assembly (524) via a fluid communication path (532).

The fluid communication path (532) may be configured such that the fluid communication path (532) may be easily separated and reconnected with one or both of the fluid handling assemblies (514,524). Similarly, in versions where an electrical communication path (530) is present, the electrical communication path (530) may be configured such that the electrical communication path (530) may be easily separated and reconnected with one or both of the controllers (512,522).

Thus, in some versions, the fluid handling subsystem (520) may be easily separated and reconnected with the instrument (510). This may be desirable to accommodate different kinds of uses of the instrument (510). For example, some uses of the instrument (510) may warrant additional fluid treatment functionality provided via the fluid treatment subsystem (520), as will be described in detail below, in which case a user may wish to couple the fluid treatment subsystem (520) with the instrument (510). Other uses of the instrument (510) may not warrant additional fluid treatment functions provided via the fluid treatment subsystem (520); in such a case, the user may wish to separate the fluid handling subsystem (520) from the instrument (510).

As an example of how the fluid handling assembly (514,524) may be used together, a set of reagents may be transferred from the fluid handling assembly (524) to the processing chip (516) via the fluid handling assembly (514) and the fluid communication path (515,532). These reagents may be processed together via a processing chip (516) to form a composition. In some such scenarios, one or more other reagents residing on the fluid handling assembly (514) (e.g., in a vial supported by a structure such as the reagent storage rack (107)) may be combined with one or more reagents from the fluid handling assembly (524) on the processing chip (516). The resulting composition may ultimately be transferred back to the fluid handling assembly (524) via the fluid handling assembly (514) and the fluid communication path (515,532). The composition may then be retrieved from the fluid handling assembly (524) for further processing. Alternatively, the fluid handling assemblies (514,524) may be used together in any other suitable manner.

In the system (500) of fig. 6, the instrument (510) and the fluid handling subsystem (520) are provided as separate components that may be removably coupled together. In some scenarios, it may be desirable to integrate all of the features and functions of the instrument (510) and the fluid handling subsystem (520) into a single instrument. To this end, fig. 7 shows an example of a system (550) that includes a single instrument (560) that removably receives a processing chip (566) that may be configured and operated as any of the variations of the processing chip (111) described herein. The example instrument (560) includes a controller (562), a first fluid handling assembly (564), and a second fluid handling assembly (570). The controller (562) may be configured and operate as the other controllers (121,512,522) described herein. The controller (562) is coupled with the first fluid processing assembly (564) via a first electrical communication path (563); and coupled with the second fluid handling assembly (570) via a second electrical communication path (571). Each electrical communication path (563,571) may include a plurality of wires, traces, other conductive paths, wireless links, and the like. The controller (562) is thus operable to drive operation of the first fluid processing assembly (564) via the electrical communication path (563); and driving operation of the second fluid handling assembly (570) via the second electrical communication path (570). While in this example two fluid handling assemblies (564,570) share the same controller (562), other versions may provide separate controllers for fluid handling assemblies (564,570), where such separate controllers communicate with each other.

The first fluid treatment assembly (564) may be configured and operated as the fluid treatment assembly (514) described above. The second fluid handling assembly (570) may be configured and operate like the fluid handling assembly (524). The first fluid treatment assembly (564) is coupled with the second fluid treatment assembly (570) via a fluid communication path (573), which may include a plurality of tubes, other conduits, and the like. The first fluid processing assembly (564) may also be coupled with a processing chip (566) via a fluid communication path (565), which may include a plurality of tubes, other conduits, and the like. In accordance with the foregoing, the system (550) may operate like the system (500). Although in this example the second fluid handling assembly (570) is integrated into the instrument (560) rather than as part of a separate sub-assembly, some versions of the system (550) may allow the second fluid handling assembly (570) to be selectively coupled and uncoupled from the controller (562) and the first fluid handling assembly (564). In such versions, the presence of the second fluid handling component (570) may be selected by a user based on the intended use of the system (550).

Fig. 8 illustrates an example of a system (600) that may represent a variation of the system (500) and/or system (600) in the context of exemplary use. In this example, the system (600) includes a first fluid handling component (610), a processing chip (612), and a second fluid handling component (614). The first fluid handling assembly (610) may be configured and operate like the fluid handling assembly (514,564). The processing chip (612) may be configured and operated as any of the variations of the processing chip (111) described herein. The second fluid handling assembly (614) may be configured and operate like the fluid handling assembly (524,570). The fluid handling assemblies (610, 614) may be integrated into a single instrument (e.g., similar to the system (550)); or provided separately and coupled together (e.g., similar to system (500)).

The system (600) of this example also includes a tray support platform (620) in which a plurality of sample trays (630,640,650,660,670,680) are arranged in a grid on an upper surface (622) of the platform (620). Each sample tray (630,640,650,660,670,680) defines a plurality of sample wells (632,642,652,662,672,682). Each sample aperture (632,642,652,662,672,682) is configured to hold a volume of fluid. The fluid handling assembly (614) includes a plurality of fluid communication paths (634,644,654) configured to provide fluid communication from and/or to the sample wells (632,642,652,662,672,682). As will be described in detail below, the fluid handling assembly (614) is operable to move the fluid communication path (634,644,654) relative to the sample tray (630,640,650,660,670,680) to selectively communicate with the sample aperture (632,642,652,662,672,682). As will also be described in detail below, the tray support platform (620) is also movable relative to the fluid communication path (634,644,654) to enable the fluid communication path (634,644,654) to reach different sample wells (632,642,652,662,672,682).

As also shown in fig. 8, a separate vial (602) may be coupled with a fluid handling assembly (614) via a fluid communication path (604). The fluid communication path (604) is configured to provide a path for fluid communication from the vial (604) to the fluid handling assembly (614). As shown, in this example, the vials (602) are separated from the tray support platform (620). In some versions, vial (602) is integrated into an instrument that includes a fluid handling assembly (610) and a handling chip (612). For example, the vial (602) may be integrated into an assembly such as a reagent storage rack (107). Although only one vial (602) is shown, the system (600) may include more than one vial (602). Further, the vial (602) is merely one example of a fluid-containing structure that may be provided separately from the tray support platform (6200).

In an example for the system (600), the system (600) may be used to perform mRNA formulation as described above in the context of block (340) of fig. 4. In some such scenarios, the processing chip (612) of the system (600) may be configured and operate like the processing chip (400) shown in fig. 5. Each sample tray (630,640,650,660,670,680) may be dedicated to a particular purpose. For example, the sample tray (630) may be used as a collection tray such that the sample wells (632) receive encapsulated mRNA formulated on the processing chip (612). Such encapsulated mRNA (e.g., in the form of ANP) may be transferred to sample wells (632) of sample tray (630) via fluid handling assemblies (610, 614) and fluid communication paths (634). When the processing chip (612) is configured like the processing chip (400), the encapsulated mRNA can be transported from a channel like channel (452). As will be described in detail below, the fluid handling assemblies (610, 614) and the fluid communication path (634) may simultaneously transfer the encapsulated mRNA from several channels, such as channels (452), in the handling chip (612) to several corresponding sample wells (632) of the sample tray (630).

The sample tray (640) may be used as an mRNA source tray such that the sample wells (642) contain mRNA fluid for use in a compounding process on the processing chip (612). Such mRNA fluids may include a combination of mRNA and formulation buffer; and may be prepared by the IVT and purification process described above with reference to blocks (320, 330) of fig. 4. Such mRNA fluid from the sample tray (640) may be transferred to the processing chip (612) via the fluid processing components (610, 614) and via the fluid communication path (644). When the processing chip (612) is configured like the processing chip (400), mRNA fluid from the sample tray (640) can be transferred to a channel like channel (402 a). As will be described in detail below, the fluid processing assembly (610, 614) and the fluid communication path (644) may simultaneously deliver mRNA fluid from several sample wells (642) to several corresponding channels on the processing chip (612), such as channel (402 a).

The sample tray (650) may be used as a DV fluid source tray such that the sample wells (642) contain DV fluid for use in a dispense process on the processing chip (612). Such DV fluids may include DV molecules in ethanol, as described above. Such DV fluids from a sample tray (650) may be transferred to a processing chip (612) via a fluid-handling assembly (610, 614) and via a fluid-communication path (654). When the processing chip (612) is configured like the processing chip (400), DV fluids from the sample tray (640) may be transferred to a channel like channel (402 b). As will be described in detail below, the fluid processing assembly (610, 614) and the fluid communication path (654) may simultaneously transfer DV fluid from a number of sample wells (652) to a number of corresponding channels on the processing chip (612), such as channel (402 b).

The sample tray (660, 670) may be used as a source tray for the flushing fluid such that the sample aperture (662,670) contains the flushing fluid for flushing the fluid communication path (644,654). The flushing fluid may also flush the channels (402 a,402 b) and structures downstream of the channels when the processing chip (612) is configured like the processing chip (400). Such a flushing fluid may comprise a combination of water and ethanol. Alternatively, any other suitable flushing fluid may be used. In some versions, the flushing fluid in the sample tray (660) is used to flush the components of the fluid communication path (654) and channel (402 b), while the flushing fluid in the sample tray (670) is used to flush the components of the fluid communication path (644) and channel (402 a). In some other versions, the rinse fluid in the sample tray (660) is used to perform a first rinse phase of the components of the fluid communication path (644,654); and the rinse fluid in the sample tray (670) is used to perform a second rinse phase of the components of the fluid communication path (644,654). In some such versions, the flush fluid in the sample tray (660) is different than the flush fluid in the sample tray (670).

The sample tray (680) may be used to collect waste from the above-described rinsing process. The sample aperture (682) may thus receive waste fluid from the fluid communication path (634). When the processing chip (612) is configured like the processing chip (400), the sample aperture (682) may also receive waste from the channel (452) and structures upstream of the channel (452).

The vial (602) may provide a diluent (e.g., PBS solution, etc.) for use in a formulation process on the processing chip (612). Such buffer solution from the vial (602) may be transferred to the processing chip (612) via the fluid processing components (610, 614) and via the fluid communication path (604). When the processing chip (612) is configured like the processing chip (400), diluent from the vial (602) may be transferred to a channel like channel (402 c). As will be described in detail below, the fluid processing assemblies (610, 614) and the fluid communication path (604) may simultaneously transfer diluent from the vial (602) to several corresponding channels on the processing chip (612), such as channel (402 c).

In some other variations, the vial (602) may contain mRNA for use in the formulation process, while the sample well (642) may contain a buffer solution. As another variation, the vial (602) may contain DV fluid, while the sample well (652) may contain a buffer solution. As yet another variation, the buffer solution, mRNA, and DV fluids may all be contained in their own respective sample trays, such that the vial (602) may be omitted.

Fig. 9-11B illustrate examples of accessory fluid treatment assemblies (700) that may be combined with or otherwise incorporated into variations of the system (100). For example, the fluid treatment assembly (700) of fig. 9 may represent a form that may be employed by the fluid treatment subsystem (520) of fig. 6. Alternatively, the fluid treatment assembly (700) of fig. 9 may represent a form that may be employed by the fluid treatment assembly (570) of fig. 7. Alternatively, the fluid treatment assembly (700) of fig. 9 may represent a form that may be employed by the fluid treatment assembly (614) of fig. 8. Alternatively, the fluid treatment assembly (700) of fig. 9 may be combined with or otherwise incorporated into a variation of the system (100) in any other suitable manner.

The example fluid treatment assembly (700) includes a frame (710) that structurally supports the remaining components of the fluid treatment assembly (700). The fluid handling assembly (700) further includes a control circuit assembly (712) secured to one side of the frame (710) and including a number of electronic components operable to drive the motorized components of the fluid handling assembly (700). The control circuit component (712) may be considered similar to the controller (522) or at least a portion of the controller (562). The fluid handling assembly (700) further includes a pneumatic assembly (714) secured to the other side of the frame (710). The pneumatic assembly (714) includes one or more pumps, flow regulators, manifolds, and/or other components for providing pressurized air to drive the pneumatic components of the fluid handling assembly (700), as described in detail below.

The fluid handling assembly (700) further includes a first tray drive assembly (720), a second tray drive assembly (730), a head support actuation assembly (740), a plurality of sampling head assemblies (750), a plurality of tray engagement assemblies (760), and a tray support platform (770). The tray support platform (770) is configured to removably receive and support a number of sample trays (780). As will be described in detail below, the tray drive assembly (720, 730) is operable to drive the tray support platform (770) relative to the frame (710) along two dimensions in a horizontal plane to selectively position the sample tray (780) relative to the sampling head assembly (750). As will also be described in detail below, the head support actuation assembly (740) is operable to drive the sampling head assembly (750) along a vertical dimension to selectively raise and lower the sampling head assembly (750) relative to the sample tray (780). As will also be described in detail below, the tray engagement assembly (760) is operable to selectively engage the sample tray (780) during at least a portion of the vertical range of travel of the sampling head assembly (750) relative to the sample tray (780). The foregoing list of components and functions of the fluid treatment assembly (700) is not intended to be exhaustive. Other components and functions may be incorporated into the fluid handling assembly (700).

B. Examples of selectively locating features of a sample tray

As noted above, and as shown in fig. 9-11B, the fluid handling assembly (700) includes a first tray drive assembly (720) and a second tray drive assembly (730) operable to drive the tray support platform (770) relative to the frame (710) along two dimensions in a horizontal plane. The first tray driving assembly (720) includes a base (722) and a bracket (not shown). The carrier is positioned and configured to support the tray support platform (770) and slide longitudinally along the base (722). The first tray drive assembly (720) further includes a motor (726) operable to drive movement of the carrier along the base (722) and thereby drive the tray support platform (770) longitudinally along the base (722). The motor (726) may be coupled with the control circuit assembly (712) via one or more cables or the like such that the control circuit assembly (712) may control activation of the motor (736). In some versions, the first tray drive assembly (720) further includes a helical gear coupled with the motor (726); and further coupled with the nut feature of the carrier such that the helical gear and nut translate rotational motion from the motor (726) into longitudinal motion of the carrier and tray support platform (770). Alternatively, any other suitable component may be used.

Fig. 10A and 10B illustrate examples of how the first tray drive assembly (730) may drive the tray support platform (770) along the horizontal "x" dimension. Fig. 10A shows the tray support platform (770) at a first position along the "x" dimension, while fig. 10B shows the tray support platform (770) at a second position along the "x" dimension. This selective positioning of the tray support platform (770) along the "x" dimension may enable certain sample apertures (782) of the sample tray (780) to be selectively positioned relative to the sampling head assembly (750), as will be described in detail below.

The second tray drive assembly (730) includes a base (734) and a bracket (732). The bracket (734) is positioned and configured to support the first tray drive assembly (720) and slide longitudinally along the base (734). The second tray drive assembly (730) also includes a motor (736) operable to drive movement of the carriage along the base (734) and thereby drive the first tray drive assembly (720) longitudinally along the base (734). The motor (736) may be coupled to the control circuit assembly (712) via one or more cables or the like such that the control circuit assembly (712) may control activation of the motor (736). When the first tray drive assembly (720) is driven longitudinally along the base (734), the tray support platform (770) will be driven longitudinally along the base (734) along with the first tray drive assembly (720). In some versions, the second tray drive assembly (730) further includes a helical gear coupled with the motor (736); and further coupled with a nut feature of the carrier (734) such that the helical gear and nut translate rotational motion from the motor (736) into longitudinal motion of the carrier (734), the first tray drive assembly (720), and the tray support platform (770). Alternatively, any other suitable component may be used.

Fig. 10A and 10C illustrate examples of how the first tray drive assembly (730) may drive the tray support platform (770) along the horizontal "y" dimension. Fig. 10A shows the tray support platform (770) at a first position along the "y" dimension, while fig. 10C shows the tray support platform (770) at a second position along the "y" dimension. This selective positioning of the tray support platform (770) along the "y" dimension, particularly in combination with the selective positioning of the tray support platform (770) along the "x" dimension as described above, may enable certain sample apertures (782) of the sample tray (780) to be selectively positioned relative to the sampling head assembly (750), as will be described in more detail below.

As noted above, and as shown in fig. 9-11B, the fluid handling assembly (700) includes a head support actuation assembly (740) operable to drive the sampling head assembly (750) along a vertical dimension to selectively raise and lower the sampling head assembly (750) relative to the sample tray (780). The head support actuating assembly (740) includes a head support plate (742), a pair of pneumatic cylinders (744), a pair of brackets (746), and a pair of rods (748). The sampling head assembly (750) is secured to a head support plate (742), as will be described in detail below. The pneumatic cylinder (744) is rigidly secured to the frame (710) by a bracket (746). The lower end of each lever (748) is rigidly secured to a head support plate (742). The upper end of each rod (748) includes a piston (not shown) contained within a corresponding pneumatic cylinder (744).

Each pneumatic cylinder (744) is coupled to the pneumatic assembly (714) via one or more tubes or other conduits such that the pneumatic assembly (714) is operable to deliver pressurized air to the pneumatic cylinders (744) to drive longitudinal translation of the rod (748) relative to the pneumatic cylinders (744). The pneumatic assembly (714) may be coupled with the control circuit assembly (712) via one or more cables or the like such that the control circuit assembly (712) may control activation of the pneumatic assembly (714). In some versions, the pneumatic cylinders (744) are double acting cylinders such that each pneumatic cylinder (744) has a first pneumatic fitting near a lower end of the pneumatic cylinder (744) and a second pneumatic fitting near an upper end of the pneumatic cylinder (744). In some other versions, the pneumatic cylinder (744) is a single acting cylinder.

Fig. 11A and 11B illustrate examples of how the head support actuation assembly (740) may drive the sampling head assembly (750) along the vertical "z" dimension. Fig. 11A shows the head support plate (742) and sampling head assembly (750) at a first position along the "z" dimension, while fig. 11B shows the head support plate (742) and sampling head assembly (750) at a second position along the "z" dimension. In the state shown on fig. 11A, the lever (748) is in a retracted position relative to the corresponding pneumatic cylinder (744); while in the state shown in fig. 11B, the lever (748) is in an extended position with respect to the corresponding pneumatic cylinder (744). By driving the sampling head assembly (750) from the upper position shown in fig. 11A to the lower position shown in fig. 11B, the head support actuation assembly (740) is operable to selectively place the sampling head assembly (750) in fluid communication with the sample apertures (782) of the sample tray (780), as will be described in detail below.

After reaching the lower position shown in fig. 11B and achieving the desired fluid communication between the sampling head assembly (750) and the sample wells (782) of the sample tray (780), the head support actuation assembly (740) may return to the state shown in fig. 11A to allow the tray drive assembly (720, 730) to drive the tray support platform (770) along a horizontal plane relative to the frame (710) to align the different sample wells (782) with the sampling head assembly (750). The above-described movements caused by the tray drive assembly (720, 730) and the head support actuation assembly (740) may be performed any suitable number of times to provide selective fluid communication between the sampling head assembly (750) and any suitable number of sample wells (782).

Although in this example the tray drive assembly (720, 730) is driven by a motor (726,736), any other suitable kind of mechanism may be used to actuate the tray drive assembly (720, 730). For example, the tray drive assembly (720, 730) may be solenoid driven, pneumatically driven, hydraulically driven, or otherwise driven. Similarly, although in the present example the head support actuator assembly (740) is pneumatically driven by the pneumatic cylinder (744) and the lever (748), any other suitable kind of mechanism may be used to actuate the head support actuator assembly (740). For example, the head support actuation assembly (740) may be motor driven, solenoid driven, hydraulically driven, or otherwise driven.

C. Examples of features to index sample trays

It may be desirable to provide features (e.g., error proofing features) that consistently provide proper positioning of the sample tray (780) on the tray support platform (770). This may help ensure that the sample aperture (782) is properly positioned relative to the sampling head assembly (750) throughout operation of the fluid handling assembly (700). Fig. 12-14 illustrate examples of such features.

As shown in fig. 12-14, the tray support platform (770) includes a set of first indexing features (800) and a set of second indexing features (810). The tray support platform (770) is configured such that one first indexing feature (800) and three second indexing features (810) will engage the sample tray (780). Each first indexing feature (800) includes a body (802) rotatably mounted to a post (804). The body (802) includes a tangentially extending arm (806) configured to engage a corner (786) of a sample tray (780). The body (802) is configured to rotate through a range of angles of motion about the post (804). A biasing member (not shown) is configured to resiliently urge the body (802) toward the angular position shown in fig. 13A. Such a biasing member of the first indexing feature (800) may comprise a torsion spring or any other suitable component. The biasing member of the first indexing feature (800) may have a shape memory that urges the biasing member toward a particular "neutral" shape or configuration (e.g., a straight configuration). The biasing member may be deformable away from the particular neutral shape or configuration (e.g., deformed into a curled shape or configuration or otherwise deformed into a curved shape or configuration), although such deformation may induce stresses in the material comprising the biasing member. Such stress may urge the biasing member back to a neutral shape or configuration.

In the context of the body (802), the biasing member of the first indexing feature (800) may be in a neutral state in the angular position shown in fig. 13A; and is in a stress state in an angular position other than the angular position shown in fig. 13A. In some other variations, the biasing member of the first indexing feature (800) may be pre-stressed when the body (802) is in the angular position shown in fig. 13A, wherein the mechanical stop feature prevents further rotation of the body (802) in a counter-clockwise direction (in the views shown in fig. 13A-13C) from the angular position shown in fig. 13A. In such versions, the biasing member of the first indexing feature (800) may be in a first stress configuration when the body (802) is in the angular position shown in fig. 13A; and is in another stressed configuration when the body (802) is rotated clockwise (in the views shown in fig. 13A-13C) to other angular positions, such that the stress in the biasing member increases when the body (802) is rotated away from the angular position shown in fig. 13A.

Whether the biasing member of the first indexing feature (800) is in a neutral state in the angular position shown in fig. 13A or in a pre-stressed state in the angular position shown in fig. 13A, as the body (802) rotates clockwise (in the views shown in fig. 13A-13C) from the position shown in fig. 13A to the other angular position, the induced or increased stress in the biasing member of the first indexing feature (800) may urge the body (802) to rotate counterclockwise (in the views shown in fig. 13A-13C) toward the angular position shown in fig. 13A. In the event that the body (802) contacts the sample tray (780) when the biasing member of the first indexing feature (800) is in a stressed state (e.g., via the arm (806)), stress in the biasing member of the first indexing feature (800) may cause the body (802) to bear against the sample tray (780). In other words, when the body (802) contacts the sample tray (780) and the body (802) is in an angular position that is not the angular position shown in fig. 13A, the body (802) will resiliently bear against the sample tray (780) because the biasing member of the first indexing feature (800) resiliently urges the body (802) back to the angular position shown in fig. 13A. This resilient bearing of the body (802) against the sample tray (780) may in turn resiliently urge the sample tray (780) into an appropriate index position along a horizontal plane (i.e., the x-y plane), as described in detail below.

Each second indexing feature (810) includes a body (812) eccentrically and rotatably mounted to a post (814). Since the body (812) is eccentrically mounted to the post (814), the off-axis region (816) of the body (812) is laterally offset from the post (814). The body (812) is configured to rotate through a range of angles of motion about the post (814). A biasing member (not shown) is configured to resiliently urge the body (812) toward the angular position shown in fig. 13A. Such a biasing member may comprise a torsion spring or any other suitable component. As best seen in fig. 14, the body (812) also includes tapered sidewalls (818). Due to the tapering of the side walls (818), the diameter of the body (812) is smaller at the upper surface (772) of the tray support platform (770) than at the top of the body (812).

Fig. 13A-13C illustrate examples of sequences of operations of indexing features (800, 810) and sample trays (780). As indicated above, fig. 13A shows the indexing features (800, 810) in a state in which the indexing features are not engaged by the sample tray (780). To load the sample tray (780) on the tray support platform (770), the user may manipulate the first indexing feature (800) to rotate the first indexing feature (800) clockwise to the position shown in fig. 13B, thereby providing clearance for the corner (786) of the sample tray (780). In the state shown in fig. 13B, the user does not have to position the sample tray (780) at the precise index position. Once the sample tray (780) has been positioned on the upper surface (772) of the tray support platform (770), such as in the position shown in fig. 13B, the user may release the first indexing feature (800).

When the user releases the first indexing feature (800) after reaching the state shown in fig. 13B, the resilient bias in the first indexing feature (800) may urge the first indexing feature (800) counterclockwise such that the arm (806) contacts the corner (786) and thereby resiliently urges the sample tray (780) into engagement with the second indexing feature (810), as shown in fig. 13C. When the outer edge (784) of the sample tray (780) engages the second indexing feature (810), the body (812) of the second indexing feature may rotate about the respective post (814). When the state shown in fig. 13C is reached, the resilient bias applied by the first and second indexing features (800, 810) on the sample tray (780) can properly index the sample tray (780) along a horizontal plane (i.e., the x-y plane). In other words, in the indexed state shown in fig. 13C, the sample aperture (782) may be properly positioned to receive the shaft (920) of the sampling head (900) of the sampling head assembly (750), as will be described in detail below.

In addition to facilitating indexing of the sample tray (780) along a horizontal plane, the second indexing feature (810) may also help ensure that the sample tray (780) is properly seated against the upper surface (772) of the tray support platform (770). As shown in fig. 14, the tapered configuration of the side walls (818) may provide a camming action against the outer edge (784) of the sample tray (780) that pushes the bottom edge (788) of the sample tray (788) fully seated on the upper surface (772) of the tray support platform (770). Such vertical pushing of sample tray (780) in the "z" direction may further facilitate proper indexing of sample aperture (782) relative to shaft (920) of sampling head (900) of sampling head assembly (750).

In some variations, the second indexing feature (810) does not rotate about the corresponding axis (814). In some such variations, the second indexing feature (810) is rigidly fixed in the angular position shown in fig. 13C.

In this example, the tray support platform (770) is sized and configured to receive six sample trays (780). Alternatively, the tray support platform (770) may be sized and configured to receive any other number of sample trays (780). In addition, the tray support platform (770) and indexing features (800, 810) may be sized and configured to accommodate sample trays (780) having 96 sample holes (782) per sample tray (780). Alternatively, the tray support platform (770) and indexing features (800, 810) may be sized and configured to accommodate sample trays (780) having any other suitable number of sample holes (782) per sample tray (780).

D. Examples of sampling head assemblies

Fig. 15-19 show components of the sampling head assembly (750) in more detail. While the fluid handling assembly (700) of the present example has three sampling head assemblies (750), other variations of the fluid handling assembly (700) may have more or less than three sampling head assemblies (750). As shown in fig. 15, each sampling head assembly (750) of the present example includes a main body (752) and four sampling heads (900). While each sampling head assembly (750) of the present example has four sampling heads (900), other variations of sampling head assemblies (750) may have more or less than four sampling heads (900).

The body (752) is rigidly mounted to a head support plate (742) of the head support actuation assembly (740). The body (752) defines a set of chambers (754), upper slots (756), and lower openings (758). The chamber (754) is configured to house a body (950) of the sampling head (950), wherein an upper portion (952) of the body (950) passes through the upper slot (756); and wherein a lower portion (756) of the body (950) passes through the lower opening (758).

As best seen in fig. 16, each sampling head (900) includes a compression member (910), a shaft (920), a ferrule (930), a spring (940), a body (950), a pneumatic fitting (970), and a washer (980). The compression member (910) includes a body (912) having a threaded region (914) and a central passage (916). The shaft (920) is hollow and includes an open upper end (922) and an open lower end (924). The collar (930) includes a frustoconical portion (932) and an annular base (934). The spring (940) of the present example is a coil spring, although any other suitable kind of resilient member may be used.

The body (950) includes a cylindrical upper portion (952), a middle block portion (954), and a cylindrical lower portion (956). As best seen in fig. 17, the upper portion (952) defines an upper passageway (960) having an inner floor (964). A main passageway (966) extends through the remainder of the length of the main body (950) from the inner floor (964) to the lower opening (968). A lateral passage (962) extends through the intermediate block portion (954) to the main passage (966).

The spring (940) is configured to fit around the upper portion (952) of the body (950). The lower end of the spring (940) is positioned and configured to bear against the upper surface of the intermediate block portion (954). The upper end of the spring (940) is positioned and configured to bear against an upper inner surface (753) in the chamber (754) of the body (752). The lower surface of the intermediate block portion (954) is positioned and configured to abut a lower inner surface (755) in a chamber (754) of the body (752). The spring (940) resiliently urges the intermediate block portion (954) toward the lower inner surface (755) such that the lower surface of the intermediate block portion (954) resiliently bears against the lower inner surface (755). However, as will be described in detail below, the springs (940) may also compress during operation of the fluid treatment assembly (700) to allow the intermediate block portion (954) to travel upward away from the lower inner surface (755).

The pneumatic fitting (970) is configured to fit in the transverse passage (962). In some versions, the pneumatic fitting (970) provides a threaded fluid-tight fit in the lateral passage (962). The pneumatic fitting (970) is further configured to couple with a flexible tube or other conduit such that the pneumatic fitting (970) can be placed in fluid communication with the pneumatic assembly (714) via the pneumatic fitting (970). This may enable the pneumatic assembly (714) to communicate pressurized air to the main passage (966) via the pneumatic fitting (970) and the lateral passage (962), as will be described in detail below.

In this example, the gasket (980) is a resilient annular member. The gasket (980) may include rubber and/or any other suitable material. The gasket (980) is configured to rest against a lower surface (958) of the lower portion (956) of the body (950), as best seen in fig. 18. The gasket (980) is also configured to provide a fluid-tight seal against a top region of a sample aperture (782) of the sample tray (780) during operation of the fluid handling assembly (700), as will be described in detail below.

The shaft (920) is coaxially disposed within the central passage (916), the ferrule (930), and the main passage (966). As shown in fig. 18-19, the main passage (966) has a diameter that is greater than the outer diameter of the shaft (920) such that a gap (967) is defined between an inner surface of the main passage (966) and an outer surface of the shaft (920). The gap (967) extends distally to a lower opening (968). The lateral passage (962) is in fluid communication with the gap (967) such that the lateral passage (962) and the gap (967) together define a passage for pressurized air to flow from the pneumatic fitting (970) to the lower opening (968). As shown in fig. 19, the collar (930) is positioned and configured to effectively seal the upper end of the gap (967). The annular seat (934) of the collar (930) is sized and configured to seat against the floor (964) of the upper passageway (966). With the shaft (920) disposed in the collar (930) and the annular base (934) seated against the bottom plate (964), the threaded region (914) of the compression member (910) may be inserted into the upper passage (966) and rotated such that the threads of the threaded region (914) engage complementary threads formed in the upper passage (966). The compression member (910) may be rotated relative to the body (950) until the compression member (910) reaches the position shown in fig. 19, at which point the distal end of the compression member (910) may bear against the frustoconical portion (932) of the collar (930) and thereby deform the frustoconical portion (932) against the shaft (920). The compressed and deformed collar (930) may thus form a seal against the shaft (920), preventing pressurized air from escaping upward from the gap (967). Alternatively, any other suitable component may be used to seal the upper end of the gap (967).

The upper end (922) of each shaft (920) may be coupled with a corresponding flexible tube (not shown) or other fluid conduit. These flexible tubes or other conduits coupled with the upper end (922) of the shaft (920) may form a fluid communication path from the fluid handling assembly (570) to the reagent storage rack (107) of the system (100). In other words, a flexible tube or other conduit coupled with the upper end (922) of the shaft (920) may be used together as a fluid communication path (532) in an arrangement such as those shown in fig. 6; or as a fluid communication path (573) in an arrangement such as those shown in fig. 7. The lower end (924) of each shaft (920) is configured to fit into a sample aperture (782) of a sample tray (780). Thus, the lower end (924) may be used to transfer fluid from or to the sample wells (782). With the ends (922, 924) open and the shaft (920) hollow (920), the shaft (920) and a flexible tube or other conduit coupled to the upper end (922) of the shaft (920) may together form a fluid communication path, such as a fluid communication path (634,644,654) between the sample tray (780) and another fluid handling assembly, such as the fluid handling assembly (610).

E. Examples of tray engagement assemblies

As noted above, the tray engagement assembly (760) is operable to selectively engage the sample tray (780) during at least a portion of the vertical range of travel of the sampling head assembly (750) relative to the sample tray (780). As shown in fig. 20-21, each tray engagement assembly (760) includes a pair of pneumatic actuators (762) and a tray engagement plate (774). Each pneumatic actuator (762) includes a pneumatic cylinder (764) and a rod (766). The pneumatic cylinder (764) is rigidly fixed to the head support plate (742). The lower end of each rod (766) is rigidly secured to a tray engagement plate (774). The upper end of each rod (766) includes a piston (not shown) contained within a corresponding pneumatic cylinder (764). Each pneumatic cylinder (764) has an upper pneumatic fitting (770) and a lower pneumatic fitting (772). The pneumatic fitting (770,772) is coupled with the pneumatic assembly (714) via one or more tubes or other conduits such that the pneumatic assembly (714) is operable to communicate pressurized air to the pneumatic cylinder (764) to drive longitudinal translation of the rod (766) relative to the pneumatic cylinder (764). In this example, the pneumatic cylinder (764) is a double acting cylinder such that the rod (766) can be actively pneumatically driven upward relative to the pneumatic cylinder (764) and actively pneumatically driven downward relative to the pneumatic cylinder (764). In some other versions, the pneumatic cylinder (764) is a single acting cylinder.

The tray engagement plate (774) includes a horizontally extending base (776) configured to engage a sample tray (780), as described in detail below. The base (776) defines a plurality of openings (778) corresponding to the sampling heads (900). The opening (778) is sized and positioned to receive the shaft (920), the washer (980), and the lower portion (956) of the body (950).

Although in this example the tray engagement assembly (760) is pneumatically driven by a pneumatic cylinder (764) and a lever (766), any other suitable kind of mechanism may be used to actuate the tray engagement assembly (760). For example, the tray engagement assembly (760) may be motor driven, solenoid driven, hydraulically driven, or otherwise driven. In some versions, the tray engagement assembly (760) is omitted.

F. Examples of sequences of engaging sample trays

Fig. 22A to 22G show examples of sequences of operating states of the head support actuation assembly (740), the sampling head assembly (750), and the tray engagement assembly (760). Although only one sampling head assembly (750) and one tray engagement assembly (760) are shown in fig. 22A-22G, the sampling head assembly (750) and tray engagement assembly (760) of the fluid handling assembly (700) may all operate synchronously throughout the stages shown in fig. 22A-22G.

In the stage shown in fig. 22A, the head support actuation assembly (740) is in an upper position such that the sampling head assembly (750) is vertically spaced above the sample tray (780). The tray engagement assembly (760) is in a configuration in which the tray engagement plate (774) is in a lowered position relative to the sampling head assembly (750) and the tray engagement plate (774) is vertically spaced above the sample tray (780). Thus, at this stage, portions of the sampling head assembly (750) or tray engagement assembly (760) are not engaged with the sample tray (780). When the head support actuation assembly (740), sampling head assembly (750), and tray engagement assembly (760) are in the state shown in fig. 22A, the tray drive assemblies (720, 730) can be actuated as described above with reference to fig. 10A-10C to properly position certain sample apertures (782) directly beneath corresponding sampling heads (900).

Once the target sample aperture (782) is properly positioned below the corresponding sampling head (900), the head support actuation assembly (740) may be actuated to drive the sampling head assembly (750) and the disk engagement assembly (760) downward toward the sample tray (780) to achieve the state shown in fig. 22B. During the transition from the state shown in fig. 22A to the state shown in fig. 22B, the sampling head assembly (750) and the tray engagement assembly (760) may travel downward in unison together such that the tray engagement assembly (760) does not move relative to the sampling head assembly (750). Actuating the head support actuation assembly (740) to achieve the state shown in fig. 22B may include: the pneumatic assembly (714) is activated to drive pressurized air to a pneumatic cylinder (744) (which is not shown in fig. 22A-22G). Upon reaching the state shown in fig. 22B, the base (776) of the tray engagement plate (774) engages the top of the sample tray (780), thereby helping the indexing features (800, 810) to maintain the position of the sample tray (780) on the tray support platform (770) during some subsequent stages of operation.

After reaching the state shown in fig. 22B, the head support actuation assembly (740) may be further actuated to drive the sampling head assembly (750) downward toward the sample tray (780) to reach the state shown in fig. 22C. When the head support actuation assembly (740) drives the sampling head assembly (750) downward, the tray engagement assembly (760) can be actuated to maintain the vertical position of the tray engagement plate (774) relative to the sample tray (780). Thus, when the sampling head assembly (750) moves downward relative to the sample tray (780), the tray engagement plate (774) may move upward relative to the sampling head assembly (750), resulting in no vertical movement of the tray engagement plate (774) relative to the sample tray (780) during the transition from the state shown in fig. 22B to the state shown in fig. 22C. This actuation of the tray engagement assembly (760) may include activating the pneumatic assembly (714) to drive pressurized air to the lower pneumatic fitting (772) of the pneumatic cylinder (764).

When the sampling head assembly (750) transitions from the state shown in fig. 22B to the state shown in fig. 22C, the lower end (924) of the shaft (920) enters the target sample aperture (782) of the sample tray (780). In a scenario in which sample fluid (782) contains a fluid, lower end (924) may be disposed in the fluid in the state shown in fig. 22B. In some versions, the sample tray (780) includes a cover member (e.g., film, plastic sheet, foil, etc.) for preventing evaporation and/or contamination of the contents of the sample wells (782). In such versions, during the transition from the state shown in fig. 22B to the state shown in fig. 22C, the lower end (924) of the shaft (920) may penetrate such a cover member to enter the sample aperture (782). In some such versions, the lower end (924) includes a sharp point, cutting edge, or other feature that aids in penetrating the cover member over the sample aperture (782) during the transition from the state shown in fig. 22B to the state shown in fig. 22C. Such penetration assisting features are optional.

When the state shown in fig. 22C is reached, the gasket (980) of the sampling head (900) may contact the upper surface surrounding the corresponding sample aperture (782). To facilitate a fluid-tight seal at such an interface between the gasket (980) and the upper surface surrounding the corresponding sample aperture (782), the head support actuation assembly (740) may be further actuated to drive the sampling head assembly (750) downward toward the sample tray (780) to achieve the state shown in fig. 22D. When the head support actuation assembly (740) drives the sampling head assembly (750) downward, the tray engagement assembly (760) can be actuated to maintain the vertical position of the tray engagement plate (774) relative to the sample tray (780). Thus, when the sampling head assembly (750) moves downward relative to the sample tray (780), the tray engagement plate (774) may move upward relative to the sampling head assembly (750), resulting in no vertical movement of the tray engagement plate (774) relative to the sample tray (780) during the transition from the state shown in fig. 22C to the state shown in fig. 22D.

During the transition from the state shown in fig. 22C to the state shown in fig. 22D, the gasket (980) may bear against the upper surface surrounding the sample aperture (782) such that the upper surface surrounding the sample aperture (782) provides an upward opposing force against the sampling head (900). The opposing force may cause the spring (940) to compress such that the lower surface of the intermediate block portion (954) disengages the lower inner surface (755) of the body (752). In other words, during the transition from the state shown in fig. 22C to the state shown in fig. 22D, the main body (752) continues to travel downward together with the head support plate (742); while the sampling head (900) remains vertically stationary during this transition. In the state shown in fig. 22D, the compression spring (940) resiliently urges the washer (980) against the upper surface surrounding the sample aperture (782). This elastic urging of the gasket (980) against the upper surface surrounding the sample aperture (782) may facilitate a fluid-tight seal at the interface between the gasket (980) and the upper surface surrounding the sample aperture (782).

With the gasket (980) securely sealed against the upper surface surrounding the sample apertures (782) during the state shown in fig. 22D, fluid may be transferred from or to the sample apertures (782) via the shaft (920). In a scenario in which fluid is transferred from the sample bore (782) to the shaft (920), such fluid communication may be achieved by activating the pneumatic assembly (914) to transfer pressurized air to the gap (967) via the pneumatic fitting (970) and the lateral passage (962). The pressurized air may exit the sampling head (900) via a lower opening (968) and thereby pneumatically bear against the surface of the fluid contained in the sample bore (782). This pressurization of the fluid by air may drive the fluid up through the lumen defined by the shaft (920). Fluid from the sample aperture (782) may leave the upper end (922) of the shaft (920) and then travel along a flexible tube or other conduit coupled to the upper end (922) of the shaft (920) to ultimately reach a destination, such as a reagent storage rack (107), a fluid handling assembly (514), a fluid handling assembly (564), a fluid handling assembly (610), and the like.

In a scenario in which fluid is transferred from the shaft (920) to the sample aperture (782), such fluid communication may be achieved by activating a source, such as a reagent cartridge (107), a fluid handling assembly (514), a fluid handling assembly (564), a fluid handling assembly (610), etc., to drive the fluid along a flexible tube or other conduit coupled to the upper end (922) of the shaft (920). Fluid may enter the lumen of the shaft (920) via the upper end (922) and exit the shaft (920) via the lower end (924) to enter the sample aperture (782). During this process, the gap (967), the lateral passages (962), and the pneumatic fitting (970) may be vented to the atmosphere, allowing air to escape from the sample aperture (782) when fluid is deposited in the sample aperture (782).

After the desired fluid has been transferred from or to the sample well (782), the process may be reversed. As part of this reversal, the head support actuation assembly (740) may be actuated to drive the sampling head assembly (750) upward toward the sample tray (780) to achieve the state shown in fig. 22E. At the same time, the tray engagement assembly (760) may be actuated to maintain the vertical position of the tray engagement plate (774) relative to the sample tray (780). Thus, when the sampling head assembly (750) is moved upward relative to the sample tray (780), the tray engagement plate (774) can be moved downward relative to the sampling head assembly (750), resulting in no vertical movement of the tray engagement plate (774) relative to the sample tray (780) during the transition from the state shown in fig. 22D to the state shown in fig. 22E. During the transition from the state shown in fig. 22D to the state shown in fig. 22E, the spring (940) may also drive the intermediate block portion (954) back into engagement with the lower inner surface (755) of the body (752).

The head support actuation assembly (740) may continue to be actuated to drive the sampling head assembly (750) upward toward the sample tray (780) to reach the state shown in fig. 22F. At the same time, the tray engagement assembly (760) may continue to be actuated to maintain the vertical position of the tray engagement plate (774) relative to the sample tray (780). Thus, as the sampling head assembly (750) continues to move upward relative to the sample tray (780), the tray engagement plate (774) may move downward relative to the sampling head assembly (750), resulting in no vertical movement of the tray engagement plate (774) relative to the sample tray (780) during the transition from the state shown in fig. 22E to the state shown in fig. 22F. During the transition from the state shown in fig. 22E to the state shown in fig. 22F, the shaft (920) may be moved away from the sample aperture (782) such that the lower end (924) of the shaft (920) is spaced above the sample tray (780) in the state shown in fig. 22F. In versions where the cover member is disposed over the sample tray (780) and the lower end (924) has penetrated the cover member during the transition from the state shown in fig. 22B to the state shown in fig. 22C, the continued engagement between the base (776) and the sample tray (780) may help prevent friction between the shaft (920) and the cover member to cause the sample tray (780) to lift upward as the shaft (920) transitions from the position shown in fig. 22E to the position shown in fig. 22F. While fig. 22F shows the base (776) remaining engaged with the sample tray (780) after the lower end (924) has substantially moved away from the top of the sample tray (780), the base (776) may be moved away from the sample tray (780) faster in the process (e.g., immediately after the lower end (924) has moved away from the top of the sample tray (780)).

After the lower end (924) has sufficiently moved away from the top of the sample tray (780), the head support actuation assembly (740) may be actuated to drive the sampling head assembly (750) and the disk engagement assembly (760) upward away from the sample tray (780) to achieve the state shown in fig. 22G. During the transition from the state shown in fig. 22F to the state shown in fig. 22G, the sampling head assembly (750) and the tray engagement assembly (760) may travel upward in unison together such that the tray engagement assembly (760) does not move relative to the sampling head assembly (750). After reaching the state shown in fig. 22G, the tray drive assemblies (720, 730) can be actuated as described above with reference to fig. 10A-10C to properly position the other sample wells (782) directly under the corresponding sampling heads (900). The process shown in fig. 22A-22G may be repeated again until the desired number of sample wells have been processed (782).

G. Examples of methods utilizing an automated fluid delivery system

Fig. 23 illustrates an example of a method of operation that may be performed using the fluid treatment assembly (700) in any of the various systems (500,550,600) described above. In some versions, the method is performed for screening purposes to determine which combination of variables produces the most appropriate encapsulated mRNA by a formulation process on a processing chip (516,566,612) such as processing chip (400). Such variables may include, but are not necessarily limited to, reagent type, buffer composition, DV formulation, reagent concentration, reagent mass ratio, process temperature, fluid flow rate ratio, and the like. Alternatively, the method may be used for any other suitable purpose.

In the context of screening applications, the process may begin with the fluid handling assembly (700) already having one, two, or more sample trays (780) loaded with reagents securely positioned on the tray support platform (770), as described above with reference to fig. 13A-13C. The sampling head assembly (750) may be positioned over the target sample aperture (782), as shown in block (1000) of fig. 23. The positioning may include actuating the tray drive assembly (720, 730) as described above with reference to fig. 10A-10C to properly position the target sample aperture (782) directly below the corresponding sampling head (900).

With the sample aperture (782) properly positioned relative to the sampling head assembly (750), the head support actuation assembly (740) and the tray engagement assembly (760) can be actuated to engage the sample tray, as shown in block (1002) of fig. 23. The engagement may include the various operational states shown in fig. 22A-22D and described above, wherein the shaft (920) is properly positioned in the target sample aperture (782), and wherein the gasket (980) provides a fluid-tight seal with an upper surface surrounding the sample aperture (782).

Next, the process may include priming the fluid pathway on the processing chip (516,566,612), as shown in block (1004) of fig. 23. In an arrangement such as that shown in fig. 6, the priming may include activating the sampling head (900) to drive reagent fluid from the sample well (782) toward the processing chip (516) via the fluid communication path (532), the fluid processing assembly (514), and the fluid communication path (515). In an arrangement such as that shown in fig. 7, the priming may include activating the sampling head (900) to drive reagent fluid from the sample well (782) toward the processing chip (566) via the fluid communication path (573), the fluid processing assembly (564), and the fluid communication path (565). In an arrangement such as that shown in fig. 8, the priming may include activating the sampling head (900) to drive reagent fluid from the sample wells (642, 652, 782) toward the processing chip (612) via the fluid communication path (644,654) and the fluid handling assembly (610, 614). Priming may also include activating one or both of the fluid handling assemblies (610, 614) to drive reagent fluid (e.g., buffer) from the vial (602) toward the processing chip (612) via the fluid communication path (604) and the fluid handling assemblies (610, 614). In some versions, the priming process includes driving the fluid at a pressure of about 0.3 psi. Further, the priming process may include driving the fluid at a flow rate in the range of about 100 microliters per minute to about 400 microliters per minute.

In some versions, the perfusion process represented by block (1004) in fig. 23 is automated. In some such versions, after sample tray (780) has been properly engaged (as represented by block (1002) in fig. 23), sampling head (900) is automatically activated to drive reagent fluid from sample wells (782) as described above until such reagent fluid reaches a predetermined location on processing chip (516,566,612). At the same time, one or both of the fluid handling assemblies (610, 614) may be automatically activated to drive reagent fluid from the vial (602). Once the reagent fluid reaches a predetermined location on the processing chip (516,566,612), fluid communication may cease until further input is provided. In some such versions, one or more sensors (e.g., sensor (105)) are used to track fluid movement on the processing chip (516,566,612) such that the one or more sensors can communicate the presence of reagent fluid in a predetermined location to a controller (e.g., controller (121,512,522,562)); and such that the controller may then automatically cease further delivery of reagent fluid until further input is provided.

In some versions where the processing chip (516,566,612) is configured as an imaging processing chip (400), predetermined locations for monitoring automatic priming from the sample well (782) may be located along the fluid channels (402 a,402 b) such that reagent fluid flow may be at least temporarily stopped before reagent fluid flows through the first mixing chamber (430). In versions where the processing chip (516,566,612) is configured like the processing chip (400) and a separate vial (602) is used to provide buffer, predetermined locations for monitoring automatic priming from the vial (602) may be located along the fluid channel (402 c) such that buffer fluid flow may be at least temporarily stopped before buffer fluid flows through the second mixing chamber (440). In versions where the controller (e.g., controller (121,512,522,562)) automatically stops further delivery of the reagent fluid in response to the fluid reaching a predetermined position, such automatic stopping may include automatically transitioning the valves (424 a,424b,424 c) to a closed state.

In providing an automatic priming version in which the controller automatically stops further delivery of reagent fluid in the primed fluid channel (402 a,402b,402 c) until a further input is provided, such further input may include user input. For example, the controller may inform the user (e.g., via the user interface (123)) that all of the appropriate fluid channels (402 a,402b,402 c) within the processing chip (400,516,566,612) have been properly primed and then wait for user input (e.g., approval) before proceeding to a later stage of the process. As another variation, the controller may track the perfusion of all fluid channels (402 a,402b,402 c) within the processing chip (516,566,612), and then automatically continue the subsequent stages in the process after the controller has determined that all appropriate fluid channels (402 a,402b,402 c) within the processing chip (400,516,566,612) have been properly perfused.

Once the processing chip (516,566,612) has been properly primed, the process may continue with performing the formulation on the processing chip (516,566,612), as represented by block (1006) in fig. 23. The formulation process may be performed as described above with reference to blocks (340, 350) of fig. 4 to produce encapsulated mRNA (e.g., in the form of ANP). In some versions, the compounding process may be completed in less than 10 milliseconds. The fluid containing the encapsulated mRNA produced by the formulation process may be transferred to the appropriate sample wells (782) in the sample tray (780), as represented by block (1008) in fig. 23.

In an arrangement such as those shown in fig. 6, communication of the fluid containing the encapsulated mRNA with the sample wells (782) may include activating the processing chip (516), the fluid processing assembly (514), and/or the fluid processing assembly (524) to drive the fluid containing the encapsulated mRNA to the appropriate sample well (782) in the sample tray (780) via the fluid communication path (515,532) and the shaft (920). In an arrangement such as those shown in fig. 7, this communication of the fluid containing the encapsulated mRNA with the sample wells (782) may include activating the processing chip (566), the fluid processing assembly (564), and/or the fluid processing assembly (570) to drive the fluid containing the encapsulated mRNA to the appropriate sample well (782) in the sample tray (780) via the fluid communication path (565,573) and the shaft (920). In an arrangement such as those shown in fig. 8, this communication of the fluid containing the encapsulated mRNA with the sample wells (632,782) may include activating the processing chip (612), the fluid processing assembly (610), and/or the fluid processing assembly (614) to drive the fluid containing the encapsulated mRNA to the appropriate sample wells (632,782) in the sample tray (630,780) via the fluid communication path (634), which may include the shaft (920).

Although the compounding and collection phases are shown in separate boxes (1006, 1008) in fig. 23, these phases may actually overlap in time. For example, reagent fluid may be transferred from a corresponding sample well (782) in the sample tray (780) while fluid containing encapsulated mRNA is transferred to other sample wells (782) in the sample tray. In the context of an arrangement such as the arrangement shown in fig. 8, the shaft (920) of the sampling head (900) forming part of the fluid communication path (634) may be disposed in the sample bore (632), while the sampling head (900) forming part of the fluid communication path (644) is disposed in the sample bore (642); and a sampling head (900) that simultaneously forms part of the fluid communication path (654) is disposed in the sample aperture (652).

As represented by block (1008) of fig. 23, to achieve transfer of the encapsulated mRNA to the appropriate sample well (782) in the sample tray (780), the purge volume of air may be transferred by: a shaft (920) that has been used to collect reagent fluid; and other fluid communication components downstream of these reagent collection shafts (920), including corresponding passages in the processing chip (400,516,566,612). In some versions of arrangements such as those shown in fig. 8, this purging may be accomplished after the reagent fluid has been evacuated from the sample aperture (662,672) such that further delivery of pressurized air via the pneumatic fitting (970) will eventually reach the shaft (920) of the sampling head (900) forming part of the fluid communication path (644,654). Pressurized air may flow through these reagent collection shafts (920) and other fluid communication components downstream of these reagent collection shafts (920) to eventually exit the shaft (920) of the sampling head (900) forming part of the fluid communication path (634).

After the fluid containing the encapsulated mRNA has been transferred to the appropriate sample wells (782) in the sample tray (780) (including the air purge described above), the process may then include flushing the reagent pathways within the fluid handling assembly (700), as represented by block (1010) of fig. 23. As part of the flush procedure, the fluid handling assembly (700) may be actuated to travel through the operational states shown in fig. 22E-22G as described above, and then the tray drive assembly (720, 730) is actuated to properly position the sample wells (782) containing the flush fluid under the corresponding sampling heads (900). In the context of the arrangement shown in fig. 8, this may include positioning the sample apertures (662) of the sample tray (660) below the corresponding sampling heads (900) of the fluid communication path (644); sample apertures (672) of sample tray (670) are positioned below corresponding sampling heads (900) of fluid communication path (654).

Once the sample aperture (782) containing the flushing fluid is properly positioned relative to the sampling head (900), the fluid handling assembly (700) can be actuated to travel through the operational states shown in fig. 22A-22D as described above to place the shaft (920) in sealed fluid communication with the flushing fluid. The pneumatic fitting (970) may be pressurized as described above to drive the flushing fluid through: a shaft (920) that has been used to collect reagent fluid; and other fluid communication components downstream of these reagent collection shafts (920), including corresponding passages in the processing chip (400,516,566,612). Thus, the flushing fluid may flush the reagent collection shafts (920) as well as other fluid communication components downstream of the reagent collection shafts (920). In the context of an arrangement such as that shown in fig. 8, the rinse fluid may be collected via the shaft (920) of fluid communication paths (634,644) that have been used to collect reagents during the priming and formulation phases represented by blocks (1004, 1006) of fig. 23.

During the flush process represented by block (1010) of fig. 23, waste fluid generated by the flush may be transferred to dedicated sample wells (782) in the sample tray (780). For example, in the system (600) described above, the sample tray (780) may be designated as a waste sample tray (680) such that the sample aperture (682) is dedicated to receiving waste. In the arrangement of the system (600) shown in fig. 8, the shaft (920) of the sampling head (900) forming part of the fluid communication path (644) may be disposed in the sample bore (682) while the sampling head (900) forming part of the fluid communication path (634) is disposed in the sample bore (672); and a sampling head (900) that simultaneously forms part of the fluid communication path (654) is disposed in the sample bore (662). Thus, the sample well (682) can easily receive waste fluid via the fluid communication path (634), while the flush fluid is transferred from the sample well (662,672) via the fluid communication path (644,654).

After an appropriate volume of rinse fluid has been delivered through the reagent collection shaft (920) and other fluid communication components downstream of the reagent collection shaft (920), these fluid passages may be dried, as represented by block (1012) of fig. 23. The drying process may include delivering pressurized air through the reagent collection shaft (920) and other fluid communication components downstream of the reagent collection shaft (920), including corresponding passages in the processing chip (400,516,566,612). In some versions of arrangements such as those shown in fig. 8, this drying may be accomplished after the flushing fluid has been evacuated from the sample aperture (662,672) such that further delivery of pressurized air via the pneumatic fitting (970) will eventually reach the shaft (920) of the sampling head (900) forming part of the fluid communication path (644,654). Pressurized air may flow through these reagent collection shafts (920) and other fluid communication components downstream of these reagent collection shafts (920) to eventually exit the shaft (920) of the sampling head (900) forming part of the fluid communication path (634). The pressurized air may flow for any suitable duration to achieve the desired dry state.

Once drying has been completed, the controller (e.g., controller (121,512,522,562)) can determine whether there are additional sample wells (782) from which to aspirate reagent, as represented by block (1014) of fig. 23. If there are additional sample wells (782) from which to draw reagent, the process may provide for positioning of the sampling head assembly (750) over the next set of target sample wells (782), as shown in block (1000) of FIG. 23. The above-described phase, represented by block (1000,1002,1004,1006,1008,1010,1012,1014), may be repeated until there are no additional sample wells (782) from which reagents are aspirated.

Once there are no additional sample wells (782) from which to draw reagent, the process may alert the user that all reagents have been used, as represented by block (1016) of fig. 23. In some versions, the alert may include an audible alert, such as a beep or other audible notification. Additionally or alternatively, the alert may include a visual alert, such as an illumination light, a graphical and/or text message on a user interface (e.g., user interface (123)), or other visual notification. In versions where the system (500,550,600) is coupled to a network, the alert may include a text message, an email message, or other type of message that is transmitted to the user over the network. Alternatively, any other suitable kind of user alert may be provided. In some versions, the user alert is omitted. Thus, the user alert represented by block (1016) of fig. 23 is optional.

After the foregoing stages have been completed, the user may retrieve the fluid containing the encapsulated mRNA and perform a test to determine the suitability of the encapsulated mRNA, as represented by block (1018) of fig. 23. In the context of the arrangement shown in fig. 8, this may include removing the sample tray (630) from the tray support platform (620) and then retrieving the fluid containing the encapsulated mRNA from the sample well (632). In some other versions, fluid containing encapsulated mRNA is retrieved from the sample well (632) prior to removing the sample tray (630) from the tray support platform (620). While in the present example the fluid handling assembly (700) is configured to deposit fluid containing encapsulated mRNA in the sample well (782), other variations may deposit fluid containing encapsulated mRNA in other types of containers (e.g., vials, etc.).

As part of the analysis represented by block (1018) of fig. 23, the user may analyze the fluid containing the encapsulated mRNA to determine various properties of the encapsulated mRNA and the fluid in which the mRNA is contained. Such properties may include, but are not necessarily limited to, encapsulation efficiency, particle size distribution, zeta potential, in vitro bioactivity, in vivo bioactivity, biodistribution in an animal (e.g., in a target organ), toxicity, stability, and the like. In some variations, other components of the fluid handling system (700) and/or system (500,550,600) include one or more integral features operable to perform at least some analysis on a fluid comprising encapsulated mRNA. For example, the instrument (510, 560) may include a dynamic light scattering station operable to detect particle size and particle distribution in a fluid containing the encapsulated mRNA.

In versions where the instrument (510, 560) and/or other components of the system (500,550,600) include one or more features that may perform an automated analysis of a fluid containing encapsulated mRNA, the system (500,550,600) may be further configured to provide real-time adjustment of delivery of reagents to the processing chip (516,566,612) in response to the results of such testing. In other words, the integrated test feature may be used to provide a feedback loop that allows the controller (512,522,562) of the system (500,550,600) to attempt to improve the compounding process to produce more desirable results.

In some scenarios, a system (500,550,600) such as those described above is operable to perform the above-described process and generate 96 discrete samples of fluid containing encapsulated mRNA in sample wells (782) in a sample tray (780) in less than two hours. In some cases, this total processing time may be significantly faster than would otherwise be required to generate a similar number of samples of fluid containing encapsulated mRNA using a system like the system (100) without the use of an accessory fluid processing component (524,570,614,700).

In this example, all sample wells (782) containing reagents contain reagents of the same formulation, so that the above procedure can be used to perform 96 tests of the same formulation procedure using the same formulation input. In some other versions, different sample wells (782) contain reagents of different formulations so that these different formulations can be tested by the procedure described above. Although the sample trays (780) of the present example each have 96 sample wells (782), the sample trays (780) may alternatively have more or less than 96 sample wells (782). Although system (500,550,600) is described above in the context of performing a screen for mRNA formulation process, system (500,550,600) may be used in any other suitable variety of processes.

VI. Miscellaneous items

The previous description is provided to enable any person skilled in the art to practice the various configurations described herein. While the subject technology has been described in detail with reference to various figures and configurations, it should be understood that these are for illustrative purposes only and should not be taken as limiting the scope of the subject technology.

Many other ways of implementing the subject technology are possible. The various functions and elements described herein may differ from those shown without departing from the scope of the subject technology. Various modifications to these implementations will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other implementations. Accordingly, many changes and modifications may be made to the subject technology by one of ordinary skill in the art without departing from the scope of the subject technology. For example, a different number of given modules or units may be employed, different types of given modules or units may be employed, given modules or units may be added, or given modules or units may be omitted.

Some versions of the examples described herein may be implemented using a processor, which may be part of a computer system, that communicates with a plurality of peripheral devices via a bus subsystem. Versions of the examples described herein implemented using a computer system may be implemented using a general purpose computer programmed to perform the methods described herein. Alternatively, versions of the examples described herein implemented using a computer system may be implemented using a special purpose computer constructed from hardware arranged to perform the methods described herein. The versions of the examples described herein may also be implemented using a combination of at least one general purpose computer and at least one special purpose computer.

In versions implemented using a computer system, each processor may include a Central Processing Unit (CPU), a microprocessor, an Application Specific Integrated Circuit (ASIC), other types of hardware components, and combinations thereof of the computer system. The computer system may include more than one type of processor. Peripheral devices of a computer system may include storage subsystems (including, for example, memory devices and file storage subsystems), user interface input devices, user interface output devices, and network interface subsystems. The input devices and output devices may allow a user to interact with the computer system. The network interface subsystem may provide an interface to external networks, including interfaces to corresponding interface devices in other computer systems. The user interface input device may comprise a keyboard; pointing devices such as a mouse, trackball, touch pad, or tablet; a scanner; a touch screen incorporated into the display; audio input devices such as speech recognition systems and microphones; as well as other types of input devices. Generally, use of the term "input device" is intended to include all possible types of devices and ways of inputting information into a computer system.

In versions implemented using a computer system, the user interface output device may include a display subsystem, a printer, a facsimile machine, or a non-visual display such as an audio output device. The display subsystem may include a Cathode Ray Tube (CRT), a flat panel device such as a Liquid Crystal Display (LCD), a projection device, or some other mechanism for creating visual images. The display subsystem may also provide for non-visual displays, such as audio output devices. Generally, the use of the term "output device" is intended to include all possible types of devices and ways to output information from a computer system to a user or to another machine or computer system.

In versions implemented using a computer system, the 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 are typically executed by a processor of a computer system, either alone or in combination with other processors. The memory used in the storage subsystem may include a plurality of memories including a main Random Access Memory (RAM) for storing instructions and data during program execution and a Read Only Memory (ROM) in which fixed instructions are stored. The file storage subsystem may provide persistent storage for program and data files, and may include a hard disk drive, a floppy disk drive along with associated removable media, a CD-ROM drive, an optical disk drive, or a removable media cartridge. Modules implementing certain embodied functions may be stored by the file storage subsystem, in a storage subsystem, or in other machines accessible by a processor.

In a form implemented using a computer system, the computer system itself may be of various 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 group of loosely networked computers, or any other data processing system or user device. Due to the ever-changing nature of computers and networks, the examples of computer systems described herein are intended only as specific examples for purposes of illustrating the disclosed technology. Many other configurations of computer systems having more or fewer components than those described herein are possible.

As an article of manufacture, rather than a method, a non-transitory 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 onto 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 disclosed methods.

The underlined and/or italicized headings and subheadings are used for convenience only, do not limit the subject technology, and are not involved in 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 are intended to be encompassed by the subject technology. Furthermore, nothing disclosed herein is intended to be dedicated to the public regardless of whether such disclosure is explicitly recited in the above description.

It is to be understood that all combinations of the foregoing concepts and additional concepts discussed in more detail below (assuming such concepts are not mutually inconsistent) are considered to be part of the inventive subject matter disclosed herein. In particular, all combinations of claimed subject matter appearing at the end of this disclosure are considered part of the inventive subject matter disclosed herein.

Claims

1. A system, comprising:

a chip receiving part for receiving a processing chip having a microfluidic channel;

a first fluid handling assembly for delivering fluid to a microfluidic pathway of a processing chip received by the chip receiving component;

a second fluid handling assembly, the second fluid handling assembly comprising:

a sample support feature for supporting a sample container, and a plurality of sampling heads for selectively delivering fluid from the sample container supported by the sample support feature; and

a fluid communication path including a plurality of conduits for providing fluid communication between the first fluid handling assembly and the plurality of sampling heads, the first fluid handling assembly for further transferring fluid from the fluid communication path to a microfluidic pathway of a processing chip received by the chip receiving member.

2. The system of claim 1, further comprising: an instrument having a housing within which the chip receiving component and the first fluid handling assembly are positioned.

3. The system of claim 2, the second fluid handling assembly positioned within the housing.

4. A system according to any one of claims 1 to 3, further comprising: a first controller for driving operation of the first fluid handling assembly.

5. The system of claim 4, further comprising: a second controller for driving operation of the second fluid handling assembly.

6. The system of claim 5, the second controller in communication with the first controller.

7. The system of any one of claims 4 to 6, the first controller to drive operation of the second fluid handling assembly.

8. The system of any one of claims 1 to 7, the chip receiving component comprising a seat mount.

9. The system of any one of claims 1 to 8, the first fluid handling assembly comprising a reagent storage rack.

10. The system of any one of claims 1 to 9, the first fluid handling assembly for storing one or more fluids.

11. The system of claim 10, the first fluid handling assembly to store one or more reagents.

12. The system of any one of claims 10 to 11, the first fluidic component for storing one or more compositions produced by a processing chip received in the chip receiving part.

13. The system of any one of claims 1 to 12, the plurality of conduits comprising a plurality of flexible tubes.

14. The system of claim 13, the flexible tube removably coupled with one or both of the first fluid handling assembly or the second fluid handling assembly.

15. The system of any one of claims 1 to 14, the sample support feature for supporting a plurality of sample trays having a plurality of sample wells.

16. The system of claim 15, the sample support feature comprising a plurality of tray indexing features for indexing the sample tray relative to the sampling head.

17. The system of claim 16, at least one of the tray indexing features for resiliently bearing against a sample tray.

18. The system of any one of claims 15 to 17, the sample support feature to support:

A first reagent sample tray for providing a first reagent, and a composition sample tray for receiving a composition formed using the processing chip received by the chip receiving part, the composition formed using the first reagent.

19. The system of claim 18, the sample support feature to support:

a second reagent sample tray for providing a second reagent, the composition sample tray for receiving a composition formed using the processing chip received by the chip receiving member, the composition formed using the first reagent and the second reagent.

20. The system of any one of claims 18 to 19, the sample support feature to support:

a flush sample tray for providing a flush fluid, and a waste sample tray for receiving waste generated by a flush process including flushing one or more of the microfluidic channels of a processing chip received by the chip receiving component.

21. The system of any one of claims 1 to 20, the second fluid handling assembly further comprising a sample support feature drive assembly for driving the sample support feature along one or more dimensions to position a sample container supported by the sample support feature relative to the sampling head.

22. The system of claim 21, the sample support feature drive assembly for driving the sample support feature along two dimensions in a horizontal plane.

23. The system of any one of claims 1 to 22, the second fluid handling assembly further comprising a head support actuation assembly for driving the sampling head to position a fluid receiving portion of the sampling head in a fluid held by a sample container supported by the sample support feature.

24. The system of claim 23, the head support actuation assembly for driving the sampling head vertically to lower and raise the sampling head relative to the sample support feature.

25. The system of any one of claims 1 to 24, each sampling head comprising:

A body defining a first passageway, an

A hollow shaft is disposed in the first passageway of the body for transferring fluid from a sample container supported by the sample support feature to the fluid communication path.

26. The system of claim 25, the first passageway having an inner diameter, the hollow shaft having an outer diameter that is less than the inner diameter such that a gap is defined between an outer surface of the hollow shaft and an inner surface of the first passageway.

27. The system of claim 26, the body further defining a lower opening in fluid communication with the gap, the body for delivering pressurized air to the lower opening via the gap.

28. The system of claim 27, each sampling head further comprising:

pneumatic fitting

A second passage, the second passage and the pneumatic fitting being in fluid communication with the gap, the pneumatic fitting and the second passage for delivering pressurized air to the gap.

29. The system of any one of claims 25 to 28, each sampling head for driving fluid from a sample container supported by the sample support feature by delivering pressurized air to an interior region of the sample container.

30. The system of claim 29, each sampling head further comprising a sealing member for engaging a portion of a sample container supported by the sample support feature.

31. The system of claim 30, the sealing member comprising an annular gasket.

32. The system of any of claims 30 to 31, further comprising: a biasing member for resiliently urging the sealing member into engagement with the portion of the sample container supported by the sample support feature.

33. The system of any one of claims 1 to 31, the second fluid handling assembly further comprising a sample container engagement assembly for selectively engaging a sample container supported by the sample support feature.

34. The system of claim 33, the sample container engagement assembly comprising:

a base

One or more actuators for selectively driving the base into and out of engagement with a sample container supported by the sample support feature.

35. The system of any one of claims 1 to 34, the sample support feature comprising a platform.

36. An apparatus, comprising:

a sample support feature for supporting a sample container;

a sampling head assembly, the sampling head assembly comprising:

mounting body

A plurality of sampling heads supported by the mounting body, each sampling head comprising:

a main body of the sampling head,

a hollow shaft supported by the sampling head body, the hollow shaft including a lower end for receiving fluid from a sample container supported by the sample support feature,

a sealing member for sealing against a surface of a sample container supported by the sample support feature, an

An opening for delivering pressurized air into a space defined above a volume of fluid in a sample container supported by the sample support feature, thereby driving the fluid from the sample container into the hollow shaft; and

a head support actuation assembly, the head support actuation assembly comprising:

a head support plate to which the mounting body is mounted, and

one or more actuators for driving the head support plate toward the sample support feature to selectively urge the lower end of the hollow shaft into a sample container supported by the sample support feature.

37. The apparatus of claim 36, further comprising: a plurality of fluid conduits for coupling the sampling head assembly with a fluid handling assembly for delivering fluid from a sample container supported by the sample support feature to a microfluidic channel in a processing chip via the fluid handling assembly.

38. The apparatus of any one of claims 36 to 37, further comprising: a sample support feature drive assembly for driving the sample support feature along one or more dimensions to position a sample container supported by the sample support feature relative to the sampling head assembly.

39. The apparatus of claim 38, the sample support feature drive assembly for driving the sample support feature along two dimensions in a horizontal plane.

40. The device of any one of claims 36 to 39, the sampling head body defining a first passageway, the hollow shaft being disposed in the first passageway of the sampling head body.

41. The device of claim 40, the first passageway having an inner diameter, the hollow shaft having an outer diameter that is less than the inner diameter such that a gap is defined between an outer surface of the hollow shaft and an inner surface of the first passageway.

42. The apparatus of claim 41, the opening of the sampling head assembly in fluid communication with the gap, the sampling head body for delivering pressurized air to the opening via the gap.

43. The apparatus of claim 42, each sampling head further comprising:

pneumatic fitting

44. The device of any one of claims 36 to 43, the sealing member comprising an annular gasket.

45. The apparatus of any one of claims 36 to 44, each sampling head assembly further comprising a resilient member for resiliently urging the seal into engagement with the surface of a sample container supported by the sample support feature.

46. The device of claim 45, the resilient member interposed between a portion of the mounting body and the sampling head body.

47. The apparatus of claim 46, the resilient member for compressing to accommodate a vertical range of motion of the sampling head body relative to the mounting body.

48. The apparatus of any one of claims 36 to 47, further comprising: a sample container engagement assembly for selectively engaging a sample container supported by the sample support feature.

49. The apparatus of claim 48, the sample container engagement assembly comprising:

a base

50. The apparatus of claim 49, the one or more actuators being secured to the head support plate.

51. The device of any one of claims 49 to 50, the base defining a plurality of openings, each opening configured to receive a corresponding hollow shaft of the plurality of sampling heads.

52. An apparatus, comprising:

a sample support feature for supporting a sample container;

a sampling head assembly, the sampling head assembly comprising:

mounting body

sampling head body

A hollow shaft supported by the sampling head body, the hollow shaft including a lower end for receiving fluid from a sample container supported by the sample support feature;

a head support plate to which the mounting body is mounted, and

one or more actuators for driving the head support plate toward the sample support feature to selectively urge the lower end of the hollow shaft into a sample container supported by the sample support feature; and

a sample container engagement assembly for selectively engaging a sample container supported by the sample support feature, the sample container engagement assembly comprising:

a base

53. The apparatus of claim 52, the one or more actuators being secured to the head support plate.

54. The device of any one of claims 52-53, the base defining a plurality of openings, each opening configured to receive a corresponding hollow shaft of the plurality of sampling heads.

55. The apparatus of any one of claims 52 to 54, further comprising: a plurality of fluid conduits for coupling the sampling head assembly with a fluid handling assembly for delivering fluid from a sample container supported by the sample support feature to a microfluidic channel in a processing chip via the fluid handling assembly.

56. The apparatus of any one of claims 52 to 55, further comprising: a sample support feature drive assembly for driving the sample support feature along one or more dimensions to position a sample container supported by the sample support feature relative to the sampling head assembly.

57. The apparatus of claim 56, said sample support feature drive assembly for driving said sample support feature along two dimensions in a horizontal plane.

58. The device of any one of claims 52 to 57, the sampling head body defining a first passageway, the hollow shaft being disposed in the first passageway of the sampling head body.

59. The device of claim 58, the first passageway having an inner diameter, the hollow shaft having an outer diameter that is less than the inner diameter such that a gap is defined between an outer surface of the hollow shaft and an inner surface of the first passageway.

60. The apparatus of claim 59, the opening of the sampling head assembly in fluid communication with the gap, the sampling head body for delivering pressurized air to the opening via the gap.

61. The apparatus of claim 60, each sampling head further comprising:

pneumatic fitting

62. The apparatus of any one of claims 52 to 61, each sampling head further comprising:

An opening for delivering pressurized air into a space defined above a volume of fluid in a sample container supported by the sample support feature, thereby driving the fluid from the sample container into the hollow shaft.

63. The apparatus of claim 62, the sealing member comprising an annular gasket.

64. The device of any one of claims 62 to 63, each sampling head assembly further comprising a resilient member for resiliently urging the seal into engagement with the surface of a sample container supported by the sample support feature.

65. The device of claim 64, the resilient member interposed between a portion of the mounting body and the sampling head body.

66. The device of claim 65, the resilient member for compressing to accommodate a vertical range of motion of the sampling head body relative to the mounting body.

67. A method, comprising:

positioning a plurality of sampling heads over a plurality of fluid containers;

inserting a hollow shaft of the sampling head into the plurality of fluid containers;

driving a first reagent from a first subset of the fluid containers via the first subset of hollow shafts toward microfluidic channels in a processing chip;

driving a second reagent toward a microfluidic channel in the processing chip;

combining the first reagent and the second reagent via the processing chip to form a composition; and

the composition is driven from the processing chip to a second subset of the fluid receptacles via a second subset of the hollow shafts.

68. The method of claim 67, the second reagent contained in a third subset of the fluid receptacles, the driving the second reagent toward a microfluidic channel in the processing chip comprising: the second reagent is driven from the third subset of the fluid containers via the third subset of the hollow shafts.

69. The method of any one of claims 67 to 68, further comprising: driving a buffer toward a microfluidic channel in the processing chip, the combining the first reagent and the second reagent via the processing chip to form a composition comprising: combining the buffer with the first reagent.

70. The method of any one of claims 67-69, the first agent comprising mRNA.

71. The method of any one of claims 67 to 70, wherein the second agent comprises a delivery vehicle.

72. The method of any one of claims 67-71, the composition comprising an encapsulated mRNA.

73. The method of claim 72, the encapsulated mRNA comprising mRNA encapsulated in a delivery vehicle molecule having nanoparticle geometry.

74. The method of any one of claims 67 to 71, further comprising: the microfluidic channels in the processing chip are perfused.

75. The method of claim 74, the priming comprising:

monitoring a target area in the microfluidic channel in the processing chip,

detecting the presence of a fluid in the target area, an

Movement of fluid in the microfluidic channel is prevented in response to detecting the presence of fluid in the target region.

76. The method of any one of claims 67 to 75, further comprising: the hollow shaft of the sampling head is removed from the plurality of fluid containers.

77. The method of claim 77, further comprising

Combining the plurality of fluid containers with a base prior to inserting the hollow shaft of the sampling head into the plurality of fluid containers; and

after removing the hollow shaft of the sampling head from the plurality of fluid containers, releasing the plurality of fluid containers from the base.

78. The method of any one of claims 67 to 77, further comprising: after driving the composition from the processing chip to the second subset of the fluid containers via the second subset of the hollow shafts, the microfluidic channels in the hollow shafts and the processing chip are rinsed.

79. The method of claim 78, further comprising: after rinsing the hollow shaft and the microfluidic channels in the processing chip, the hollow shaft and the microfluidic channels in the processing chip are dried.

80. The method of any one of claims 78 to 79, further comprising: a flushing waste fluid is collected in a third subset of the fluid containers, the flushing waste fluid being generated by flushing the hollow shaft and the microfluidic channels in the processing chip.

81. The method of any one of claims 67 to 80, further comprising: determining whether another subset of the fluid containers contains more of the first reagent.

82. The method of claim 81, further comprising:

determining that a third subset of the fluid containers contains more of the first reagent;

positioning a plurality of sampling heads over the third subset of fluid containers;

inserting a hollow shaft of the sampling head into the third subset of fluid containers;

driving a first reagent from the third subset of the fluid containers via the first subset of the hollow shafts toward microfluidic channels in a processing chip;

driving a second reagent toward a microfluidic channel in the processing chip;

combining the first reagent and the second reagent via the processing chip to form a subsequent composition; and

the subsequent composition is driven from the processing chip to a fourth subset of the fluid containers via the second subset of the hollow shafts.

83. The method of any one of claims 81 to 82, further comprising:

determining that another subset of the fluid containers does not contain more of the first reagent; and

Alerting the user to the completion of the composition forming process.

84. A processor readable medium comprising content for causing a processor to process data by performing the method of any one of claims 67 to 83.

85. A system according to any one of claims 1 to 35, comprising: the device of any one of claims 36 to 66.