Systems, devices, and methods for bioprocessing

Abstract

Systems, devices, and methods for automated bioprocessing are provided. A system for automated bioprocessing includes a workcell having a feedthrough and at least one instrument for processing RNA, and a cartridge for processing and configured to interface with the at least one instrument. The cartridge includes a transcription module and a purification module, and temperature within at least a portion of each of the transcription and purification modules is maintained by a temperature control system of the instrument. The system also includes a pump for transferring the material through the transcription and purification modules, and a robotic arm for transferring the cartridge between the feedthrough and the at least one instrument.

Description

SYSTEMS, DEVICES, AND METHODS FOR BIOPROCESSINGCROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Patent Application No. 63 / 675,215 filed July 24, 2024, the content of which is incorporated herein by reference in its entirety for all purposes.TECHNICAL FIELD

[0002] Systems, devices, and methods herein relate to manufacturing mRNA therapeutics for biomedical applications using automated systems.BACKGROUND

[0003] Messenger RNA (mRNA) is utilized in cancer treatments to stimulate a patient’s immune system against cancer cells. These treatments, which include vaccines and other therapies, some of which may be deemed “off-the-shelf’, are tailored for individual patients or groups based on the mutational profile of their cancerous tumors. In particular, diseaserelevant proteins, or neoantigens, produced by cancer cells are different from those produced by non-cancer cells. The immune system recognizes these neoantigens as foreign and will mount a response against any cell expressing these proteins. Accordingly, mRNA can be used as a treatment (e.g., vaccine) to provide a single-stranded molecule that encodes neoantigens that are likely to trigger an immune response for a particular patient or group of patients. The increased demand for both bulk (one batch treats many patients) and individualized (one batch treats one patient) mRNA therapeutics and vaccines requires a technology platform and cost-effective manufacturing process with a well-defined product characterization.

[0004] mRNA processing begins with transcribing the mRNA using linearized plasmid DNA as a template. Second, the mRNA is purified to remove unreacted reagents and other impurities. Third, special carriers such as lipid nanoparticles (LNPs) are provided with the mRNA to form a particle capable of entering cells. Fourth, and finally, the mRNA undergoes sterile filtration to ensure the sterility of the final mRNA product. The resulting mRNA product can be used to produce specific target antigens in cells and trigger an immune response.

[0005] Conventional mRNA processing is largely manual, which is costly, limits repeatability, reliability, and throughput of these processes, and risks the sterility of samples throughout processes. Manual handling introduces variability between batches, increases therisk of operator error, and requires extensive personnel training and gowning procedures for cleanroom operations. The open handling of materials during transfers between processing steps presents opportunities for microbial contamination and cross-contamination between different products. Furthermore, manual processes typically require larger cleanroom facilities with higher operational costs for maintaining appropriate environmental conditions. The labor-intensive nature of current methods also limits the ability to rapidly scale production in response to pandemic threats or to cost-effectively produce batches for personalized medicines.

[0006] Accordingly, there is a need for systems and methods that provide automated, end- to-end processing of mRNA therapeutics in a scalable way to meet the growing demand.SUMMARY

[0007] The present disclosure relates generally to systems, devices, and methods for automatic bioprocessing.

[0008] An automated system for bioprocessing may include a workcell with a feedthrough and at least one instalment for processing nucleic acid material, a cartridge for processing the nucleic acid material, and a robotic arm for transferring the cartridge between the feedthrough and the at least one instrument. The cartridge may be configured to interface with the at least one instrument. The cartridge may include a transcription module and a purification module. A temperature within at least a portion of each of the transcription and purification modules may be maintained by a temperature control system of the at least one instrument. The cartridge may further include a pump for transferring the material through the transcription and purification modules. In some variations, the workcell may have an enclosure configured to maintain a sterile environment from a beginning to an end of the bioprocessing. The sterile environment may be an ISO8 cleanroom. In some variations, the cartridge may be a first cartridge, and the system may further include a second cartridge for processing nucleic acid material. The system may be configured to process the nucleic acid material of each of the first and second cartridges in parallel. In some variations, workcell may further include a reagent management system with a storage compartment for storing reagents. A temperature of the storage compartment may be configured to be maintained within a desired range. Additionally, the reagent management system may have an interior zone that is separate from an interior zone of the workcell. In some variations, the feedthrough may be a first feedthrough, and the robotic arm may be configured to transfer the cartridge to and from thereagent management system via a second, different feedthrough. In some variations, the workcell may further include an analytical instrument system with one or more analytical instruments for determining one or more parameters of an RNA product of the system. The one or more parameters may include one or more of a percent purity, a percent yield, and a yield of the RNA product. In some variations, the system may further include a controller communicably coupled to the at least one instrument and the robotic arm. Tire controller may be configured to generate an electronic batch record for an RNA product of the system. In some variations, the RNA product may be an mRNA product.

[0009] Another automated system for bioprocessing may include a workcell with at least one instrument for processing nucleic acid material, a feedthrough for introducing materials into the workcell, a robotic arm for transferring materials within the workcell, and a cartridge for processing the nucleic acid material. The cartridge may be configured to interface with the at least one instrument. The cartridge may include a transcription module, a purification module, an integrated pump for transferring the nucleic acid material between and through the transcription and purification modules, and interfaces for thermal regulation by the at least one instrument. Tire at least one instrument may include a temperature control system configured to maintain a temperature within at least a portion of each of the transcription and purification modules of the cartridge, and one or more mechanical interfaces for engaging with the cartridge. The robotic arm may be configured to transfer the cartridge between the feedthrough and at least one instrument.

[0010] A cartridge for automated bioprocessing may include a plurality of modules, each module configured to interface with at least one instrument of a workcell to process nucleic acid material, a fluidic bus for transferring fluid through the plurality of modules, and a tube configured to engage a pump for driving fluid flow through the fluidic bus. The plurality of modules may include a transcription module, a purification module, and a particle formation module. In some variations, a temperature of one or more of the transcription, purification, and particle formation modules may be controlled by the at least one instrument. In some variations, the cartridge may further include a digestion module, and the transcription and digestion modules may have one or more shared components. The one or more shared components may include a fluid compartment. In some variations, the purification module may include one or more of a tangential flow filtration module, a magnetic separation module, and a chromatography module. In some variations, the cartridge may further include a final formation module configured to store a final product of the nucleic acid materialhaving RNA. In some variations, the cartridge may further include a waste module with a first fluid compartment for storing aqueous waste and a second fluid compartment for storing organic waste. In some variations, the cartridge may further include a storage module with a fluid compartment for storing a reagent. The fluid compartment may be configured to interface with a thermal element of the at least one instrument, and wherein the thermal element is configured to control a temperature of the reagent within the fluid compartment. In some variations, the cartridge may have a housing with one or more ports for transferring samples of the nucleic acid material to an analytical instrument during processing. In some variations, the cartridge may be a single-use cartridge.

[0011] Another cartridge for automated bioprocessing may include an enclosure comprising a plurality of processing modules, a fluidic bus fluidically coupling the plurality' of modules, one or more ports for fluid transfer, a plurality of reservoirs for storing reagents, buffers, and waste, one or more external interfaces configured to engage with a bioprocessing instrument, and one or more sensors for monitoring process parameters. The plurality of modules may be configured to perform a series of bioprocessing operations. The one or more external interfaces may be configured to facilitate one or both of energy and material transfer between the cartridge and the instrument. The cartridge may be configured to produce nucleic acid-based products.

[0012] An instrument for automated bioprocessing may include a receiving bay configured to receive and support a cartridge carrying material for RNA processing, and a plurality of actuating modules. Each module may be configured to drive one or more modules of the cartridge to process the material, and the plurality of actuating modules may include a transcription actuation module configured to actuate a transcription reaction within the cartridge. The instrument may further include a temperature control system configured to maintain a temperature within the one or more modules of the cartridge. The instrument may be housed within an enclosed workcell. In some variations, the instrument may further include a first thermal control element configured to control a first temperature of a transcription module of the one or more modules of the cartridge, and a second thermal control element configured to control a second temperature of a purification module of the one or more modules of the cartridge. The first thermal control element may be a component of the transcription actuation module. In some variations, the instrument may further include a purification actuation module, and the second thennal control element may be a component of the purification actuation module. In some variations, the instrument may further includeone or more sensors configured to monitor a parameter of the cartridge throughout the RNA processing. The one or more sensors may include one or more bubble sensors, and the parameter may be a fluid level within the one or more modules of the cartridge. The one or more sensors may be configured to transmit real-time measurements of the parameter to a controller of the workcell. In some variations, the instrument may further include a pump configured to engage one or more fluid conduits of the cartridge to pump fluid through the cartridge.

[0013] Another instrument for automated bioprocessing may include a housing structure configured to maintain controlled environmental conditions, a receiving bay configured to receive and interface with a self-contained bioprocessing cartridge, a plurality of actuating modules configured to engage with corresponding modules of the cartridge to perform an automated bioprocessing workflow, a temperature control system configured to maintain a plurality of discrete thermal zones of the cartridge, a fluid transfer system for transferring materials to and from said cartridge, and one or more sensors for real-time process monitoring and quality assessment. The instrument may be configured to perform the automated bioprocessing workflow in conjunction with the cartridge to produce a finished nucleic acid therapeutic product.

[0014] A method for automated bioprocessing may include providing a workcell having one or more instruments, a feedthrough, and a robotic arm. Via the robotic arm, the method may include transferring a first cartridge from the feedthrough to a first of the one or more instruments, where the first cartridge may be configured to process a first nucleic acid material, and transferring a second cartridge from the feedthrough to a second of the one or more instruments, where the second cartridge may be configured to process a second nucleic acid material. Both of the first and second cartridges may include at least a transcription module and a purification module. The method may further include processing the first and second nucleic acid materials in parallel. In some variations, processing the first and second nucleic acid materials may include performing a series of bioprocessing steps on each of the first and second nucleic acid materials. The sequence of bioprocessing steps may include a transcription step, a digestion step, a purification step, a particle formation step, and a final formulation step. In some variations, processing the first and second nucleic acid materials may include performing one or more analysis steps on each of the first and second nucleic acid materials. Performing the one or more analysis steps may include determining one or more of a yield, a percent yield, and a percent purity of the first and second nucleic acidmaterials. In some variations, the method may further include, prior to transferring the first and second cartridges to the first and second instruments, respectively, sterilizing the first and second cartridges within the feedthrough. In some variations, the method may further include transferring the first and second cartridges from the first instrument and the second instrument, respectively, to the feedthrough after the first and second nucleic acid materials are processed.

[0015] Another method for automated bioprocessing may include transferring a cartridge from a feedthrough of a workcell to an instrument of the workcell via a robotic arm. The cartridge may include a transcription module and a purification module, and the instrument may be configured to perform one or more processing steps on material within the cartridge. The method may further include actuating the transcription module to enable a transcription reaction therein, determining a yield of the transcription reaction, and, when the yield is about equal to or greater than a threshold, transferring the material from the transcription module to the purification module. In some variations, determining the yield may include transferring a sample of the material to an analytical tool. Transferring the sample may include removing the sample from the cartridge via a port thereon. The analytical tool may be located in the cartridge, in the instrument, or in an analytical instrument system of the workcell. In some variations, when the analytical tool is within the analytical instrument system, transferring the sample may include moving the cartridge to the analytical instrument system via the robotic arm. In some variations, the method may further include, prior to transferring the material to the purification module, transferring the material to a digestion module. In some variations, the method may further include, prior to transferring the material to the purification module, performing a digestion processing step on the material within the transcription module.BRIEF DESCRIPTION OF THE DRAWINGS

[0016] FIG. 1 A is a block diagram of an illustrative variation of a workcell of an automated bioprocessing system. FIG. IB is a block diagram of an illustrative variation of a cartridge of the automated bioprocessing system. FIG. 1 C is a block diagram of an illustrative variation of a control system of the automated bioprocessing system. FIG. ID is a block diagram of an illustrative variation of a reagent management system of the automated bioprocessing system. FIG. IE is a block diagram of an illustrative variation of an analytical instrument system of the automated bioprocessing system.

[0017] FIG. 2 depicts an illustrative variation of an automated bioprocessing system including a workcell with an instrument and a cartridge.

[0018] FIG. 3 is a schematic diagram of an illustrative variation of a cartridge for automated bioprocessing.

[0019] FIG. 4 is a flow diagram of an illustrative variation of a method for automated bioprocessing using systems and devices herein.

[0020] FIG. 5 is a flow diagram of an illustrative variation of a conditional execution method for automated bioprocessing.

[0021] FIG. 6 is a flow diagram of an illustrative variation of a method for automated mRNA processing.DETAILED DESCRIPTION

[0022] Systems, devices, and methods for integrated and automated processing for biomedical applications are described herein. These systems are designed to fully automate mRNA processing through closed systems that execute end-to-end processing of therapeutics, from initial materials to final product. The systems address both personalized products for individual patients and larger-scale batch products for multiple patients, and may be configured to process a plurality of products in parallel to maximize throughput. The systems are suitable for development, pre-clinical, clinical, or GMP manufacturing, with the capability to operate unit operations in parallel or series as needed.

[0023] RNA therapeutics, particularly mRNA products, hold significant promise in addressing various medical needs. They offer new therapeutic avenues in combatting diseases like cancer, multiple sclerosis, heart disease, and infectious diseases. mRNA vaccines have been developed for influenza, COVID- 19, HIV, Zika, Lyme disease, among others, with ongoing research for additional applications. mRNA products are also pivotal for gene editing and protein replacement therapies. Compared to vaccines utilizing viral delivery systems, mRNA vaccines demonstrate superior safety profiles, versatility, and simplified processing via templated methods.

[0024] Despite their promise, current mRNA processing solutions remain labor-intensive, time-consuming, and prone to human error and contamination, making scalability challenging. Meeting the increasing demand for mRNA therapeutics necessitates automated, integrated systems to reduce labor costs, contamination risks, and processing times while enhancing scalability. The systems described herein address these challenges by integratingunit operations within a closed environment, providing real-time process monitoring and control, and maintaining comprehensive electronic batch records for regulatory compliance - all while reducing both capital and operational costs compared to traditional manufacturing approaches.

[0025] In some variations, the systems may comprise a workcell housing bioprocessing instruments and a cartridge carrying biomaterials (e.g., DNA templates for mRNA transcription, intermediate mRNA products, and / or final mRNA products). The cartridge may be inserted into the workcell via a feedthrough and may be sterilized within the workcell using an integrated sterilization system. The cartridge may include modules for various bioprocessing steps such as transcription, digestion, capping, tailing, purification, particle formation (e.g., lipid nanoparticle formation), concentration, and filling. Each instrument may incorporate actuating modules configured to interface with specific modules within the cartridge, providing the necessary actuating, electrical, magnetic, mechanical, thermal, and / or fluidic interactions to facilitate each step of the bioprocessing workflow. The workcell may further be equipped with integrated systems like reagent management, waste management, and analytical instruments for comprehensive processing and analysis. A materials handling system may facilitate transfer of the cartridge and processing reagents between feedthroughs, instruments, and integrated systems while maintaining sterile conditions. The workcell may also include a control system configured to coordinate operations, manage data, and oversee processing.

[0026] Manufacturing personalized mRNA therapeutics presents unique challenges that these systems are specifically designed to address. Individualized mRNA neoantigen therapy, for example, requires patient-specific mRNA sequences based on each patient's unique tumor mutational profile. This demands a manufacturing process that combines automation and sterility with exceptional flexibility for small, customized batches. The system must rapidly switch between different patient-specific sequences without cross-contamination while maintaining consistent quality. Time-sensitive cancer treatment requirements add pressure for swift turnaround from tumor sequencing to vaccine delivery. These personalized medicines also require sophisticated analytical and quality control processes to verify each unique mRNA sequence and its encapsulation. Traditional economies of scale don’t apply to patientspecific production runs, necessitating novel approaches to cost-effective single or limiteddose batch manufacturing. To meet these scalability requirements, the systems and methodshere may be configured to produce multiple batches in parallel, achieving the high throughput necessary for personalized medicine production.

[0027] To accommodate these diverse processing needs, the cartridges herein may comprise a variety of sizes. Smaller cartridges may suit personalized mRNA therapeutics (e.g., individualized neoantigen cancer treatments) for individual patients, while larger cartridges may be configured to contain batches for treating many patients with the same therapeutic. The bioprocessing instruments herein may be correspondingly sized to match these cartridges. Furthermore, components of both the cartridge modules and instrument actuating modules herein may be customized or replaced, enabling tailored bioprocessing workflows.

[0028] As used herein, the terms “material”, “biomaterial”, and “genetic material”, “sample”, and “product” refer to an initial, intermediate, or final product of a bioprocessing workflow. The material may include one or more biomolecules, such as proteins, lipids, nucleic acids, carbohydrates, and combinations thereof. For mRNA processing, material may include nucleic acid-based material, such as one or more of DNA, a DNA template, and RNA (e.g., crude mRNA, unpurified and / or purified mRNA), including portions thereof. Additionally, or alternatively, in some variations, the material may include tRNA, rRNA, taRNA, derivatives thereof, and / or combinations thereof. As discussed herein throughout, the material may be carried and processed by a cartridge throughout a bioprocessing workflow. In some variations, one or more samples of the material may be taken from the cartridge and analyzed. Such samples may or may not be returned to the cartridge following analysis.

[0029] As used herein, the terms “module” and “actuating module” refer to a set of one or more components or tools of a device (e.g., a cartridge or an instrument) that are used to perform a bioprocessing operation. In some variations, the modules and / or actuating modules herein may be reconfigurable. In some variations, the modules and / or actuating modules herein may be used to perform more than one bioprocessing operation.

[0030] Detailed exemplary variations of these systems, devices, methods, and aspects thereof are elaborated below.Automated Bioprocessing System

[0031] hi general, the automated bioprocessing systems herein may comprise a workcell housing one or more bioprocessing instruments, and a cartridge carrying biomaterial for processing. The cartridge may be loaded into the workcell through a feedthrough. Theworkcell may include a sterilization system operably coupled to the feedthrough and configured to sterilize the cartridge. The cartridge may include one or more modules for performing various bioprocessing steps. For example, the cartridge may include one or more of a transcription module, a digestion module, a capping & tailing module, a purification module, a particle formation module, and the like. Each instrument may include one or more corresponding actuating modules configured to drive the one or more cartridge modules to carry out a bioprocessing workflow (or at least a portion thereof). The systems may produce RNA therapeutics, such as mRNA vaccines, using templates of synthesized DNA and / or DNA sampled from one or more patients.

[0032] The workcell may include additional integrated systems such as a reagent management system (RMS) for storing common reagents, a waste management system (WMS) for collecting processing waste, and an analytical instrument system (AIS) for analyzing the product throughout the workflow. A materials handling system (MHS) may be configured to move the cartridge between the feedthrough, the instruments, and the integrated systems. The MHS may also be configured to transfer fluids (e.g., reagents, waste, and product samples) between the cartridge and the integrated systems. The fluids may be transferred throughout the workcell (e.g., via one or more robots of the MHS) in fluid devices to maintain a sterile processing environment. Further, the workcell may include a control system to coordinate instrument and system operations, as well as manage data collection, analysis, and storage during processing. Additionally, the control system may be configured to generate one or more electronic batch records for a sample throughout its workflow.

[0033] The use of the MHS (e.g., one or more robots) and control system may enhance the automation, efficiency, and sterility of the systems herein. In some variations, the workcell may be configured to process a plurality of cartridges, and thus a plurality of products, in parallel. For example, the workcell may include a plurality of instruments, each instrument independently configured to perform one or more operations upon material (e.g., genetic material, such as DNA and / or RNA) within the cartridge.

[0034] The systems may produce biotherapeutics for use in treating patients, such as mRNA to be administered as a vaccine to one or more patients. The therapeutic products may be stored in bulk or in distinct doses, either of which may be frozen. In some variations, the therapeutic products may undergo one or more post-processing operations, such as lyophilization, to maintain and stabilize the product during storage, distribution, etc.

[0035] In some variations, the cartridges herein may be provided in various sizes to accommodate different scales of therapeutic dose production. For example, a first cartridge having a total fluid capacity on the order of microliters (pL) to milliliters (mL) may be configured to process small batches of therapeutics. Such a cartridge may be useful in processing personalized therapeutics (e.g., for cancer treatments) for a single patient or a small group of patients (e.g., fewer than 10 patients, fewer than 25 patients, fewer than 50 patients, fewer than 75 patients, or fewer than 100 patients). In contrast, a second cartridge having a total fluid capacity on the order of liters (L) may be configured to process relatively larger batches of therapeutics, such as for vaccine production for many individuals (e.g., more than 10 patients, more than 25 patients, more than 50 patients, more than 75 patients, more than 100 patients, more than 500 patients, more than 1,000 patients, more than 2,500 patients, or more than 5,000 patients). Accordingly, the first cartridge may be smaller than the second cartridge in one or more dimensions. The bioprocessing instruments of the workcell may also be provided in various sizes (e.g., a first size and a second, larger size) to accommodate different cartridge sizes. In some variations, the systems herein may include a first set of one or more cartridges having a first fluid capacity and a second set of one or more cartridges having a second, larger fluid capacity. The systems may additionally include a first set of one or more instruments configured to interface with the first set of cartridges, and a second, larger (in size - height, width, and / or length) set of one or more instruments configured to interface with the second set of cartridges. In some embodiments, the instruments may be of similar size, but may include built-in features (e.g., clamping features) that can accommodate cartridges of different sizes.

[0036] Additionally, in some variations, components of the cartridge modules and / or instrument actuating modules may be updated, removed, or replaced to provide a uniquely configured cartridge and / or instrument. To this end, highly customized bioprocessing workflows may be achieved using the automated bioprocessing systems and devices herein.

[0037] FIG. 1 A is a block diagram of an illustrative variation of an automated bioprocessing system (“system”) 100 comprising a workcell 110 and a cartridge 160. The workcell 110 and the cartridge 160 may be components of a modular system 100 that facilitates end-to-end processing of therapeutic product (e.g., RNA). That is, the product may be completely processed on the cartridge 160 within the workcell 110. In some embodiments, the workcell 110 may be designed to perform one or more unit operations as part of a development, pre-clinical, clinical, or GMP manufacturing process.

[0038] In some variations, the workcell 110 may be configured to process a plurality of cartridges 160 in parallel. The processing and parallel processing operations described herein may be designated and controlled by the control system 140. Specifically, the control system 140 may actuate one or more components of the workcell 110 to execute a processing workflow for a product. The control system 140 may also be configured to monitor and adjust the processing workflow based on real-time feedback, as is discussed herein.I. Workcell

[0039] The workcell 110 may comprise an enclosure 101 defining an interior zone 103. The enclosure 101 may comprise folly, or at least partially, enclosed housing inside which one or more bioprocessing operations may be performed in a fully, or at least partially, automated process. In some variations, the enclosure 101 may be configured to meet International Organization for Standardization (ISO) standard ISO8 or better (e.g., ISO7 or ISO6). An advantage of meeting ISO8 or better standards is that the system 100 may be used in a facility that does not meet ISO8 standards (i.e., that lack a clean room or other sufficiently filtered air space). Optionally, the facility' may be an ISO8 or ISO9 facility. In some variations, a workcell may comprise a volume of less than about 800 m3, less than about 700 m3, less than about 600 m3, less than about 500 m3, less than about 300 m3, less than about 250 m3, less than about 200 m3, less than about 150 m3, less than about 100 m3, less than about 50 m3, less than about 25 m3, less than about 10 m3, and less than about 5 m3, including all ranges and sub-values in-between.

[0040] The workcell 110 may include a feedthrough 106, which may be positioned within a side wall of the enclosure 101. In some variations, the workcell 110 may include a plurality of feedthroughs 106. Within the interior zone 103, the workcell 110 may include a robot (e.g., a first robot) of a materials handling system (MHS) 104 and instruments 120. The workcell 110 may also include reagent management system (RMS) 180 and an analytical instrument system (AIS) 190. In some variations, one or both of the RMS 180 and the AIS 190 may be separated from the interior zone 103 of the workcell by a wall or partition. The RMS 180 may be configured to store, prepare, and transfer processing reagents to the instruments 120 and / or to the AIS 190, as explained herein below. The AIS 190 may comprise one or more analytical instruments for analyzing a therapeutic product or intermediate thereof during a bioprocessing workflow. The MHS 104 may include one or more second robots, such as a second robot within an interior zone of the RMS, for moving reagents, product samples,waste (e.g., within fluidic devices) throughout the workcell 110. The workcell 110 may further include systems for monitoring and controlling workcell parameters and operations throughout processing. These systems may include one or more of an environmental monitoring system (EMS) 108, sterilization system 112, and a waste management system (WMS) 114. To enable fully automated bioprocessing within the workcell 110, a control system 140 may be communicably coupled to each of the robot 104, the feedthrough 106, the instruments 120, the EMS 108, the sterilization system 112, the WMS 113, the RMS 180, and the AIS 190 such that a network is formed for the system 100.1. Materials Handling System (MHS)

[0041] The MHS 104 may include one or more robots communicably coupled to and actuated by the control system 140. The MHS 104 may be configured to move cartridges and / or fluidic devices (carrying reagents, samples, waste) through the workcell 110. A robot of the MHS 104 may include one or more robotic arms, such as one, two, at least one, at least two, or more than two robotic arms. In some variations, the MHS 104 may comprise a first robotic arm configured to operate within the interior zone 103 of the workcell 110, and a second robotic arm configured to operate within an interior zone of the RMS 180 (or within an interior zone of a feedthrough coupled to the RMS 180). Each robotic arm may be translatable along at least one axis and / or rotatable around at least one axis. For example, a robotic arm may be coupled to and moved throughout the workcell 110 via a track or rail (e.g., linear rail) positioned centrally (e.g., on a floor or ceiling) within the workcell 110.Additionally, or alternatively, a robotic arm may comprise one or more pivot joints (e.g., one, two, three, more than three, etc.) around which at least a portion of the robotic arm may rotate. Further, each robotic arm may comprise one or more end effectors configured to precisely handle the cartridge 160 and / or a fluidic device. For example, an end effector may be configured to releasably couple ports (e.g., fluid transfer ports) on the cartridge 160 and / or the fluidic device to corresponding ports on the fluidic device and / or the cartridge 160 to enable (sterile ) fluid transfer between the cartridge 160 and the fluidic device. In some variations, the end effector may be temperature-controlled for carefid handling of sensitive reagents. The MHS 104, such as the end effector thereof, may include one or more sensors, such as a scanner and / or a camera, configured to track each product throughout its processing (e.g., via identification, such as an RFID tag, or label, on the cartridge carrying the product).2. Feedthrough

[0042] The feedthrough 106 may be configured as a window through which cartridges and / or fluidic devices are moved in and out of the interior zone 103 of the workcell 110. In some variations, the workcell 110 may include a plurality of feedthroughs (e.g., two or more feedthroughs). For example, a first feedthrough may be configured to receive cartridges for processing, and a second feedthrough may be configured to pass fluidic devices (e.g., fluid containers, including sterile liquid transfer devices) carrying processing reagents, product samples, and / or waste between an interior zone 103 and an interior zone of the RMS 180. The feedthrough 106 may comprise a receiving bay for supporting the cartridge and / or or other fluidic device therein. The feedthrough 106 may extend through a side wall of the workcell. The sidewall may face the interior zone 103 of the workcell at a first end, and may face either an external environment or an interior zone of the RMS 180 at a second, opposite end.

[0043] The feedthrough 106 may comprise one or more sensors, such as a seamier and / or a camera, configured to identify each product at the start and end of its processing workflow (e.g., via identification, such as an RFID tag, on the cartridge carrying the product).Similarly, the one or more sensors may be used to identify reagents being exchanged (e.g., via identification, such as an RFID tag, on the fluidic device carrying the reagent) and / or wastes being collected throughout processing.

[0044] The feedthrough 106 may be configured to sterilize a cartridge and / or fluidic device using ultraviolet radiation (UV), or chemical sterilizing agents provided as a vapor, spray, or wash. The feedthrough 106 may optionally be configured to automatically and / or periodically spray, wash, irradiate, or otherwise treat cartridges and / or fluidic devices (e.g., with ethanol and / or isopropyl alcohol solutions, vaporized hydrogen peroxide (VHP)) during processing to maintain sterility of the interior zone 103 of the workcell 110 (e.g., ISO 7 or better ).

[0045] Referring briefly to FIG. 2, bioprocessing system 200 is shown to include workcell 210 and cartridge 260. The workcell 210 may comprise an enclosure 201 having at least one feedthrough 206 having receiving bay 207. The interior dimensions of the receiving bay 207 may be about equal to or greater than corresponding exterior dimensions of the cartridge 260. While FIG. 2 may depict the feedthrough 206 on a particular enclosure sidewall of the workcell 210, it should be understood that the feedthrough 206 may be provided on any suitable enclosure sidewall of the workcell 210. hi some variations, more than one feedthrough 206 may be provided on a single enclosure sidewall of the workcell 210. The robot 204 may be configured to retrieve the cartridge 260 from the receiving bay 207 at thebeginning of a bioprocessing protocol, and to unload the cartridge 260 within the receiving bay 207 after processing.3. Environmental Monitoring System (EMS)

[0046] Turning back to FIG. 1A, the EMS 108 may include one or more sensors configured to monitor one or more environmental conditions of the interior zone 103, including, but not limited to, temperature, humidity, particle counts, and particle concentrations. The monitoring may be continuous, periodic, or may occur upon request (e.g., by the control system 140). In some variations, the measured environmental parameter may be compared to a range of acceptable values. If the measurement is outside of its designated range, control system 140 may automatically alert an operator. Such a notification may be provided on a workcell and / or RMS-specific display.4. Sterilization System

[0047] The sterilization system 112 may comprise one or more of a sterilant source, fluid source, and a pump. One or more components of the system 100, such as the cartridge 160 and / or other fluidic devices, may be sterilized and / or aerated by circulating one or both of a sterilant and fluid (e.g., heated air, vaporized hydrogen peroxide (VHP)) via the sterilization system 112. In some variations, the sterilization system 112 may comprise one or more of vaporized hydrogen peroxide (VHP), UV light, electron-beam (e-beam) sterilization, dry thermal decontamination, and steam-in-place. In general, the sterilant or decontaminant may be configured to sterilize any external surface of any component of the workcell 110. At a first end, the sterilant and / or fluid source may be connected to the pump and to one or more areas within the workcell. For example, the sterilant and / or fluid source may be fluidically and / or electrically connected to the pump, and may be connected (e.g., via tubing) to one or more of the feedthrough 106, the RMS 180 (e.g., to a feedthrough and / or storage compartment thereof), and the AIS 190 (e.g., at least a portion thereof). In some embodiments, with respect to sterilizing fluidic devices within the RMS 180 (discussed below), for fluidic devices carrying temperature-sensitive reagents, the sterilant may include VHP, while for fluidic devices carrying reagents configured to be stored at room-temperature, the sterilant may include UV-C light.

[0048] The control system 140 may control one or more sterilization parameters. For example, the control system may be communicably coupled to the pump to control a rate offluid and / or sterilant flow throughout the workcell 110. In some variations, the control system 140 may control a sterilant / fluid delivery duration. Additionally, or alternatively, the sterilization system 112 may comprise one or more sensors for monitoring sterilization parameters such as, for example, the sterilant / fluid flow rate, the sterilant / fluid delivery duration, a stored sterilant / fluid amount (e.g., mass, volume), and the like.

[0049] In some variations, the sterilization system may provide a sterility assurance level (SAL) of at least 10-3 SAL.5. Reagent Management System (RMS)

[0050] The RMS 180 may be configured to store, sterilize, stabilize, prepare, and transfer commonly used reagents for bioprocessing, such as for RNA processing. Tire reagents may include one or more genetic materials (e.g., DNA templates), lipids, buffer, cytokines, proteins, enzymes, polynucleotides, nutrients, cryoprotectants, solvents, cellular materials, and pharmaceutically acceptable excipients. Some reagents may be sensitive to conditions like temperature, humidity, and / or light. For example, nucleotide triphosphates (NTPs) may be sensitive to freeze-thaw cycles, and may be best preserved at temperatures of less than 0°C (e.g., for long term storage). Enzymes like RNA polymerase, DNase, RNA triphosphatase, guanylyltransferase, and methyltransferase, poly(A) polymerase, etc., as well as cap analogs and modified nucleotides, may similarly require storage at temperatures of less than 0°C. The activity of these enzymes may be maintained when stored as single-use aliquots within the RMS 180. To store DNA templates, a humidity-controlled refrigerated storage compartment (2-8°C) may be an ideal environment for storing DNA templates. Storage temperature requirements for Lipid components for RNA-lipid particle formation, such as lipid nanoparticles (LNP) may depend on a particle type.

[0051] A block diagram of an illustrative variation of the RMS 180 is depicted in FIG. ID. As shown, the RMS 180 may define an interior zone 181 within which storage compartments 182 are housed. The storage compartments 182 may be temperature-controlled, as described in detail below. The interior zone 181 may be separated from the interior zone 103 of the workcell 110 by a wall or partition, which may comprise a feedthrough for transferring reagents between the two zones. One or more sensors 186 may be configured to monitor one or more environmental conditions of the interior zone, including, but not limited to, temperature, humidity, particle counts, and particle concentrations. The monitoring may be continuous, periodic, or may occur upon request. In some variations, the measuredenvironmental parameter may be compared to a range of acceptable values. If the measurement is outside of its designated range, the control system 140 may automatically alert an operator. Such a notification may be provided on a workcell 110 and / or RMS- specific display.

[0052] The RMS 180 may also include fluidic devices 184. In general, reagents for bioprocessing may be maintained in the fluidic devices 184, which may enable sterile fluid transfer therein and out via one or more ports. In general, the fluidic devices 184 herein may have a volume of about 1 mL to about 1 ,000 mL to accommodate various reagent volumes. Additionally, each fluidic device 184 may comprise a unique barcode or RFID such that the reagent therein may be tracked throughout storage, preparation, and processing. Further, in some variations, the fluidic devices 184 may comprise one or more integrated sensors for monitoring a volume and / or mass of the reagent therein. Such data may be used (e.g., by the control system 140 ) to determine when to refill a fluidic device, and / or to monitor fluid transfer in real time. The fluidic devices 184 herein may be reusable or may be configured for single-use.

[0053] The MHS 104 may be configured to move the fluidic devices throughout the interior zone. As detailed above, a robot of the MHS 104 may have barcode and / or RFID reading capabilities to validate chain of custody and chain of command for all fluidic devices. The robot may be configured to transfer the fluidic devices to and from an RMS -specific feedthrough, as discussed above.

[0054] The RMS 180 may include one or more storage compartments (e.g., a plurality thereof) of for the fluidic devices. The storage compartments may comprise drawers, cabinets, vaults, carousels, and / or the like. A storage compartment may have a storage capacity of about 5 L to about 250 L, such as about 10 L to about 200 L, about 15 L to about 150 L, about 20 L to about 100 L, about 25 L to about 75 L, or about 30 L to about 50 L, including all ranges and subranges therebetween (e.g., about 20 L, about 50 L, about 100 L, or about 200 L).

[0055] In some variations, storage compartments of varying capacities may be utilized. Moreover, the storage compartments may be temperature-controlled storage compartments to stabilize the reagents stored therein. That is, a predetermined temperature (or acceptable range of temperatures) may be monitored and controlled for each storage compartment. A storage capacity of a storage compartment may depend on an amount of reagent requiring temperature control within a temperature range of the storage compartment. In somevariations, a temperature range of a storage compartment may be adjustable. In some variations, the temperature to be maintained may be set as a threshold temperature and when the temperature deviates from the threshold temperature, a control system 140 may be engaged to try to maintain the threshold temperature. In some embodiments, the threshold temperature may be within a temperature range (e.g., between about 2 degrees C and about 8 degrees C). In some variations, the temperature range may be between about 4 degrees C and about 8 degrees C, about 3 degrees C and about 5 degrees C, about 2 degrees C and about 6 degrees C, about 0 degrees C and about 10 degrees C, about 0 degrees C and about 12 degrees C, about 0 degrees C and about 14 degrees C, about 0 degrees C and about 16 degrees C, about -2 degrees C and about 10 degrees C, about -2 degrees C and about 8 degrees C, about -4 degrees C and about 10 degrees C, about -4 degrees C and about 8 degrees C, or about -4 degrees C and about 14 degrees C. In some variations, the storage compartment may be configured to maintain a temperature of about 4 degrees C.

[0056] When the threshold temperature cannot be maintained, the control system 140 may be engaged to provide one or more alerts (e.g., audible, visual, etc.) to an operator of the workcell 110 or to one or more displays of the workcell 110. The threshold temperature may set to less than about 4 degrees C, such as about 3 degrees C, about 2 degrees C, about 1 degree C, about 0 degrees C, about -2 degrees C, about -4 degrees C, about -6 degrees C, about -8 degrees C, or colder. The storage compartment may be operatively coupled to a sensor system such that a measurement measured by the sensor system may determine a response by the storage compartment.

[0057] hi some examples, a freeze storage compartment may be maintained at about equal to or less than 0°C (and may have, e.g., about 20L to about 50 L capacity). A refrigerated storage compartment may be maintained at a temperature of between about 0°C and about 10°C, such as about 2°C to about 8°C. This storage compartment may have about 50 L to about 200 L capacity. Finally, room temperature storage compartment may be maintained at a temperature of between about 15°C to about 25°C. This storage compartment may have about 100 L to about 200 L capacity.

[0058] In some variations, one or more conditions within a storage compartment 184 may be monitored and controlled. Non-limiting examples of such conditions may include temperature, humidity, and / or brightness within the storage compartment. For example, the storage compartment may comprise one or more sensors (e.g., temperature, humidity, and / or light sensors) configured to detect (continuously, periodically, or upon request) a storagecompartment parameter, and provide the data to the control system in real-time. The control system 140 may analyze the feedback (e.g., compare the measurement(s) to a parameter threshold or condition) and adjust the parameter appropriately. In some variations, the measured storage compartment parameter may be compared to a range of acceptable values. If the measurement is outside of its designated range, the control system 140 may automatically alert an operator. Such a notification may be provided on a workcell and / or RMS-specific display.

[0059] Further, the RMS may comprise a reagent preparation station (RPS) 188. The reagent preparation station 188 may be an area within the interior zone 181 of the RMS where a reagent may be prepared for immediate use in processing. The RPS 188 may include automated tools configured to perform one or more reagent preparation steps. For example, a reagent may need to be thawed, heated, or cooled prior to use to maximize its potency. Accordingly, the RPS 188 may include thermal control elements configured to interface with the fluidic device carrying the reagent and provide precise temperature control thereto. The thermal control elements may be digitally controlled by the control system 140. As another example, a particularly sensitive reagent may be lyophilized so that it may be stored at ambient temperatures. To prepare the lyophilized reagent for use, the RPS 188 may be configured to automatically reconstitute the reagent using water or buffer. Furthermore, the RPS may comprise one or more mixing chambers, which may be temperature-controlled (e.g., via thermal elements) to facilitate buffer preparation, and LNP formation. A stir rate within the one or more mixing chambers may be controlled by the control system 140.

[0060] hi some variations, the RMS 180 may not be utilized for storing some or all of the reagents for bioprocessing. For example, reagents for enzymatic reactions (e.g., one or more of RNA polymerase, DNase, RNA triphosphatase, guanylyltransferase, and methyltransferase, poly(A) polymerase, etc.) may be stored outside of the workcell 110, while the temperature, moisture, and light-sensitive reagents may be stored within the temperature controlled storage units.

[0061] In some variations, the RMS 180 may include a controller that is communicably coupled to but distinct from a central controller of the control system 140. The RMS controller may include a data management system configured to provide real-time inventory management, reagent usage monitoring and forecasting, automated reordering of reagents, and / or generation of detailed audit trails. In some variations, the RMS may include a user interface (e.g., touch screen display) for monitoring and, in some cases, operating one ormore components of the RMS 180. In some variations, the controller may employ 21 CFR Part 11 compliant software.

[0062] In some variations, the RMS may include the waste management system (WMS) 114, which may comprise fluidic devices and / or storage compartments for storing waste produced during cartridge processing.6. Analytical Instrument System (AIS)

[0063] The AIS may comprise one or more analytical instruments for analyzing a sample. FIG. IE is a block diagram of an illustrative variation of the AIS 190. As shown, the AIS 190 may have an interior zone 192 within with analytical instruments 194 may be stored. In some variations, a cartridge may be transferred from the interior zone 103 of the workcell 110 to the interior zone 192 of the AIS 190 via a feedthrough. In some variations, a fluidic device may be transferred from the interior zone 103 of the workcell or the interior zone of the RMS 180 110 to the interior zone 192 of the AIS 190 via a feedthrough.

[0064] The analytical instruments 194 may be (releasably) couplable to a cartridge and / or a fluidic device for collecting and analyzing a fluid sample therefrom. For example, an analytical instrument 194 may be connected (e.g., by a robot or by an operator) to the cartridge 160 by one or more fluidic conduits. The analytical instrument 194 may then perform an analysis to quantify and / or characterize the fluid sample, during or after which the instrument 194 may be disconnected from the cartridge 160. A sample taken for analysis may generally comprise a known volume of about 0 p F to about 300 pL of the sample being processed depending on the analytical instrument 194 used. In some variations, more than one analytical instrument 194 may be connected to the cartridge 160 at once. In some variations, one or more analytical instruments 194 may be reusable. The analytical instruments 194 may comprise one or more of a flow cytometer, a cell counter, a quantitative thermocycler (e.g., qPCR), a fluorimeter, a chromatograph (e.g., a high-performance liquid chromatograph (HPLC)), an electropherogram, a UV-Vis spectrophotometer, a dynamic light scattering (DLS) instrument, an electrophoretic light scattering (ELS) instrument, and the like. A chromatograph may include one or more columns such as a reverse-phase column (RP-HPLC), an ion pair reverse-phase column (IP-RP HPLC), an ion-exchange column, a liquid exchange column, and a liquid-solid exchange column. In some variations, the analytical instruments 194 may comprise two or more of an instrument type, such as two or more liquid chromatographic columns. Each of the analytical instruments 194 may becommunicably coupled to the control system 140 so that the bioprocessing workflow may be automatically adjusted in response to the analysis.

[0065] Products may be sampled after one or more steps of a bioprocessing workflow to determine whether adjustments should be made to the workflow. Specifically, a parameter of a resultant product (e.g., intermediate, or final product) of a given bioprocessing operation may be determined, and when the parameter does not meet a standard (e.g., threshold or condition), the workflow may be modified (e.g., to repeat at least a portion of the preceding operation) until the parameter does meet the threshold or condition. Examples of this analysis for RNA (e.g., mRNA) processing are described below.

[0066] RNA processing may include the steps of transcribing a DNA template to mRNA, digesting remaining portions of the DNA template (which may occur simultaneously as or overlap with the transcribing), purifying the DNase-treated mRNA, capping (and, in some variations, tailing) the purified mRNA, repurifying the capped and tailed mRNA, forming mRNA-loaded lipid particles, and formulating the mRNA-lipid product for fill & finish. In some variations, one or more of the processing operations may occur in the same compartment (e.g., fluidic compartment on the cartridge 160). In some variations, an RNA product may only be purified after the formulating step (e.g., only a fully processed rnRNA- LNP may be purified). Determining a product yield or purity' percentage after these operations may inform whether modifications should be made to the workflow. For example, if the measured product yield or purity percentage is below a threshold, the preceding operation may be repeated to ensure that the process results in a high-yield, high-purity mRNA product. Additionally, RNA and RNA therapeutics may be degraded by exposure to heat, hydrolysis, oxidation, light, and ribonucleases throughout manufacturing. Thus, it may be important to assess batch-to batch consistency, process repeatability, and the resulting mRNA quality during mRNA processing.

[0067] Accordingly, the analytical instruments 194 may include one or more HPLC instruments (RP-HPLC, IP-RP-HPLC, and / or other column / separation modes) for determining purity percent, yield, and / or yield percent of one or more of the crude RNA, the DNase-treated mRNA, the purified mRNA, the capped and tailed mRNA, the purified capped and tailed mRNA, the mRNA-lipid product, and the purified and concentrated mRNA-lipid product. An HPLC instrument may also be used to identify impurities within the aforementioned intermediate products. Additionally, or alternatively, the analytical instruments 194 may include one or more UV-Vis spectrophotometers for determiningmRNA concentration and purity of one or more of the crude RNA, the DNase-treated mRNA, the purified mRNA, the capped and tailed mRNA, and the purified capped and tailed mRNA. Additionally, or alternatively, the analytical instruments 194 may include one or more Raman spectrophotometers .

[0068] In some variations, a threshold purity and / or yield percent may be about 50% to about 100%, such as about 55% to about 99%, about 65% to about 95%, about 65% to about 90%, about 70% to about 85%, or about 75% to about 80% (including all ranges and subranges therebetween).7. Instruments

[0069] The instruments 120 may comprise one or more bioprocessing instalments (BPI). The BPI(s) may be configured to execute workflows for processing therapeutic products (e.g., RNA). For example, a BPI may be configured to execute one or more RNA processing steps including transcription, digestion, capping & tailing, purifying (e.g., RNA and / or RNA- lipid particle purification), and / or concentrating, hi some variations, the BPI may be configured to execute all of the aforementioned RNA processing steps. In general, a control system may be communicably coupled to an instalment and may provide instructions thereto for automatically executing a processing workflow. The instalment may be configured to engage the cartridge to carry out at least a portion of the workflow on material (e.g., biomaterial, genetic material) within the cartridge. The engagement may be achieved via electrical, mechanical, magnetic, fluidic, optical, and / or other interfaces. For example, electrical interfaces may enable the instalment to transfer power and / or data to the caaridge. As another example, mechanical interfaces, such as locks, may stabilize the cartridge within the instalment. As yet another example, magnetic interfaces may allow the instrument to drive one or more bioprocessing operations (e.g., mRNA purification by magnetic separation). In still another example, fluidic interfaces may allow the instalment to transfer fluid (e.g., reagent) to and from the cartridge. As another example, optical interfaces may allow the instrument to monitor one or more cartridge parameters, such as a current level of fluid within one or more fluid compartments thereof. Typically, an instalment may be configured to engage a single cartridge at once. Accordingly, the systems herein may include a plurality of instalments (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or more than 15 BPIs) to process a plurality of cartridges in parallel.

[0070] In some variations, an instrument 120 may be configured to automatically detect and verify a cartridge type and / or batch identification information of the cartridge introduced therein. For example, the instrument 120 may include barcode and / or RFID reading capabilities, and may transmit this information to the control system 140 to await affirmative instructions from the control system 140 to continue the bioprocessing workflow.

[0071] Discussed below are variations of instrument systems and their actuating modules, which may be used to engage and actuate corresponding modules on the cartridge. The systems may enable an instrument, such as a bioprocessing instrument (BPI), to monitor and / or control one or more process parameters throughout the processing. As used herein, the term “actuating module”, refers to a set of components (e.g., one or more components having shared and / or dissimilar functions) that are capable of engaging with a corresponding module on the cartridge to complete a particular processing step, such as a portion of an RNA manufacturing process. The modules may be actuating modules configured to drive corresponding modules of the cartridge to perform specific bioprocessing operations.

[0072] Further, discussed below are variations of instrument systems and their constituent or actuating modules designed to enable comprehensive monitoring and control of process parameters throughout the RNA manufacturing workflow. In this context, an “instrument system,” such as a BPI, refers to an integrated platform that advantageously oversees and manages the entire RNA production process by interacting with various modules of the cartridge to execute specific steps in the manufacturing workflow. The term “module” denotes a set of components designed to perform a particular processing step within the RNA manufacturing process. These modules may comprise one or more components with shared or distinct functions and can be categorized into two main types: instrument or actuating modules, which are part of the BPI or other instrument systems, and cartridge modules, which are components within the consumable cartridge. Instrument modules are designed to interface with and control corresponding cartridge modules, providing the necessary mechanical, thermal, fluidic, or other interactions to drive specific bioprocessing operations within the cartridge.

[0073] Some or all of the systems and actuating modules may be integrated in a fixed configuration within the instrument, though they need not be. For example, one or more of the actuating modules may be configurable or moveable (e.g., by an operator, controller, and / or robot of the workcell) within the instrument enclosure, permitting various formats of instruments to be assembled. That is, the instrument may have configurable or moveableactuating modules coupled by configurable fluidic, mechanical, optical, and / or electrical connections. Similarly, in some variations, the components of various actuating modules herein may be interspersed with each other such that each module may be defined by the set of connected components that collectively perform a predetermined function. However, these components of an actuating module may or may not be physically grouped within the instrument.

[0074] FIG. IE is a block diagram showing an illustrative variation of the instrument 120. The instrument 122 may include a receiving bay 122. The receiving bay 122 may be positioned on or within a sidewall of a workcell, and may be partially or fully enclosed. In some variations, the receiving bay 122 may comprise a floor, a ceiling, and one or more sidewalls (e.g., at least one, at least two, or at least three sidewalls, such as a three or four sidewalls). In some variations, the instrument 120 may include a cover, such as a door, configured to transition the receiving bay 122 between an open configuration and a closed configuration. The receiving bay 122 may be configured to receive a cartridge in the open configuration, and may be actively processing a cartridge in the closed configuration.

[0075] Referring again to FIG. 2, the bioprocessing system 200 may include the cartridge 260 and instrument 220. The instrument 220 may include receiving bay 222. The interior dimensions of the receiving bay 222 may be about equal to or greater than corresponding exterior dimensions of the cartridge 260. Within the receiving bay 222, a plurality of interfaces (e.g., electrical, mechanical, magnetic, optical, fluidic, and / or the like, not shown) enable systems and actuating modules of the instrument 220 to physically engage, detect, and / or otherwise interface with the cartridge 260. The robot 204 may move the cartridge between the instrument 120, the feedthrough 206, and other areas of the workcell 210 (e.g., the RMS and / or the AIS, as described herein) throughout a bioprocessing workflow.

[0076] Turning back to FIG. 1, to process the cartridge, the instruments 120 may include systems for controlling process parameters of the cartridge, including fluid flow (e.g., amounts, paths, rates thereof), temperature, pressure, and the like. Such systems may include the fluid transfer system 124, temperature control system 126, and / or the analytical system 128.7.1. Fluid Transfer System (FTS)

[0077] The FTS 124 may comprise one or more pumps and one or more pump actuators (e.g., a motor coupled to a rotor of a pump). These components may be coupled together aswell as coupled to an interior sidewall of the instrument 120 (e.g., mounted thereto). Each of the one or more pumps actuators (e.g., motors) may be operably coupled to the control system 140, which may control an operational speed thereof. Adjusting this operational speed may correspondingly adjust a pressure within a corresponding fluid transfer system of the cartridge (e.g., within its fluidic tubing), to cause fluid flow through the cartridge.7.2. Temperature Control System (TCS)

[0078] As discussed herein, some bioprocessing operations and reagents may require precise temperature control to function. The TCS 126 may interface with one or more thermal elements configured to contact or be positioned adjacent to a portion of the cartridge 160 that is within the instrument receiving bay 122. The portion of the cartridge 160 may comprise a module or portion thereof, such as a bottom of a fluid compartment of a cartridge module. Tire thermal elements may comprise Peltier elements and / or resistive heaters, and may be controlled by a controller of the TCS. In some variations, the thermal elements may provide heat exchange (heating and cooling) through direct contact with the cartridge 160. This exchange may be allow for temperature control that results in an actual temperature (of the portion of the cartridge) that is about 0.01 °C to about 1 °C of the target temperature, such as about 0.05 °C to about 0.75 °C, about 0.1 °C to about 0.5 °C, or about 0.15 °C to about 0.25 °C of the target temperature, (including all ranges and subranges therein). In some variations, the TCS 126 may comprise at least one thermal element in contact with at least a portion of each module of the cartridge 160. The TCS 126 may advantageously enable different modules of the cartridge to maintain different target temperatures or temperature ranges, thereby accommodating the varied thermal requirements of mRNA manufacturing. Accordingly, using the TCS 126, one or more cartridge modules may comprise a distinct temperature-controlled zone.7.3. Analytical System

[0079] The instruments 120 may include an integrated analytical system 128 comprising sensor(s) and / or analytical tool(s) for monitoring a bioprocessing workflow in real-time. In some variations, one or more components of the analytical system 128 may be communicably coupled to the control system 140 such that continuous feedback on the workflow may be analyzed by the control system 140. The control system 140 may use this information to inform subsequent operations of the workflow (e.g., to modify such subsequent operations).

[0080] In some variations, the analytical system 128 may include one or more pH sensors and / or conductivity sensors in line with one or both of the purification and particle formation modules to monitor such parameters throughout all purification and particle formation processes. In some variations, the analytical system 128 may include one or more bubble sensor and camera in line with at least one fluid compartment sidewall of the cartridge 160. The bubble sensor(s) and / or camera(s) may provide real-time feedback on cartridge parameters like a fluid level within a module.

[0081] Additionally, or alternatively, the analytical system 128 may include one or more liquid chromatographic columns for determining purity percent, yield, and / or yield percent of one or more of the crude RNA, the DNase-treated mRNA, the purified mRNA, the capped and tailed mRNA, the purified capped and tailed mRNA, the mRNA-lipid product, and the purified and concentrated mRNA-lipid product. Additionally, or alternatively, the analytical system 128 may include one or more UV-Vis spectrophotometers for determining mRNA concentration and purity of one or more of the crude RNA, the DNase-treated mRNA, the purified mRNA, the capped and tailed mRNA, and the purified capped and tailed mRNA. Additionally, or alternatively, the analytical system 128 may include one or more DLS and / or ELS instruments for characterizing one or both of a particle size and a particle size distribution of mRNA-lipid product and / or purified and concentrated mRNA-lipid product.

[0082] The instrument 120 may also include actuating modules for executing the bioprocessing operations. These modules may include the transcription actuating module 130, purification actuating module 132, and particle formation actuating module 134.7.4. Enzyme Reaction Actuating Module (ERAM)

[0083] The ERAM 130 may actuate one or more of the transcription, digestion, and capping & tailing modules of the cartridge 160. In doing so, the ERAM 130 may ensure homogenous reaction conditions within these modules (e.g., within a mixing chamber thereof). In some variations, one or more modules of the cartridge may be actuated simultaneously by the ERAM 130. Accordingly, the ERAM 130 may drive one or more of the transcription, digestion, and capping & tailing steps of mRNA manufacturing. To do so, the ERAM 130 may interface with one or more of the aforementioned modules via a magnetic coupling. In particular, a magnetic stirrer or mixing paddle within a fluid compartment of the cartridge may be drawn to the magnet of the ERAM 130. This magnet may be rotatable, and may have a variable operational speed of about 0 RPM to about 500RPM, such as about 25 RPM to about 400 RPM, about 50 RPM to about 350 RPM, about 75 RPM to about 300 RPM, about 100 RPM to about 250 RPM, or about 150 RPM to about 200 RPM (including all ranges and subranges therein). The speed of the rotatable magnet of the ERAM 130 may be controlled by the control system 140. In some variations, the rotatable magnet, via instructions from the control system 140, may be configured to apply one or more programmable mixing patterns within the transcription, digestion, and / or capping & tailing modules.7.5. Purification Actuating Module (PAM)

[0084] The PAM 132 may include one or more sub-actuating-modules for actuating corresponding submodules of the purification module of the cartridge. The sub-actuatingmodules may include one or more of a tangential flow filtration actuating module (TFFAM), a magnetic selection actuating module (MSAM), and a chromatography actuating module (CAM).

[0085] The TFFAM be configured to automatically control a pressure differential transversely across the membrane of the TFFM, and to regulate a flow rate of fluid longitudinally across the membrane of the TFFM. The TFFAM may achieve automated pressure control via a pressure regulator coupled to a compartment of the TFFM. Additionally, the TFFAM may be configured to control the diafiltration cycle during TFF by actuating the fluidic tubing of the cartridge 160 with the pump actuator of the instrument 120 to control buffer exchange.

[0086] The MSAM may comprise one or more permanent magnets mounted on a mechanism (e.g., a swivel) that is configured to engage the flowcell on the cartridge 160. Accordingly, the MSAM may enable magnetic separation used to purify an mRNA intermediate product. Additionally, the MSAM may include one or more sensors, such as bubble sensors and / or cameras, to monitor one or both of a fluid level and a flow rate within the flowcell. When the fluid level and / or flow rate is about equal to or greater than a threshold (determined by the control system 140), indicating that the flowcell may overflow, the magnetic separation process may be modified to relieve prevent such overflow. Thus, the MSAM may advantageously help prevent device and system damage by monitoring fluid transfer within the cartridge 160.

[0087] Like the TFFM, the CAM may utilize the pump actuator of the instrument 120 to deliver buffer for the chromatography module. Additionally, the CAM may operate one ormore columns of the chromatography using valves. Further, the CAM may include one or more pressure sensors configured to detect pressure within the one or more columns. When the measured pressure is about equal to or greater than a threshold (determined by the control system 140), the chromatography process may be modified to relieve the pressure therein. Thus, the CAM may help prevent device and system damage by monitoring pressure within the cartridge 160. Additionally, the TFFM may include one or more thermal elements of the TCS 126 for controlling a temperature of its corresponding cartridge module.7.6. Particle Formation Actuating Module (PF AM)

[0088] The PF AM 134 may employ the pump actuator(s), thermal elements of the TCS 126, as well as automated cleaning and flushing procedures (via, e.g., the control system 140) to maintain optimal conditions for the formation of lipid nanoparticles encapsulating mRNA within the particle formation module of the cartridge 160.IL Control System

[0089] The control system 140 may include one or more controllers for monitoring and controlling a cell processing workflow. In some variations, the control system 140 may include at least one central controller in communication with each automated component of the system 100. As such, the controller may be configured to control and / or manage one or more bioprocessing workflows being carried out within the system 100. In some variations, the controller may simultaneously control a plurality of bioprocessing workflows being carried out on samples of a corresponding plurality of bioprocessing cartridges.

[0090] In some variations, the control system 140 may be configured to perform a conditional execution method where data may be received from one or more sensors of the systems and devices herein, the data may be compared to a threshold or condition, and when the threshold or condition is not met, the control system 140 may adjust the corresponding bioprocessing workflow to optimize for the parameter or condition (e.g., to achieve a higher product yield).

[0091] In some variations, the control system 140 may include a data management system configured to retrieve data from the RMS 180 to accomplish real-time inventory management, reagent usage monitoring and forecasting, automated reordering of reagents, and / or generation of detailed audit trails.

[0092] Referring to FIG. IB, the control system 140 (e.g., controller or computing device) of the workcell 110 may include one or more of a processor 142, memory 144, communication device, 146, input device 148, and display 150. A processor of the system controller (e.g., processor 142) may process data and / or other signals to control one or more components of the system. The processor 142 may be configured to receive, process, compile, compute, store, access, read, write, and / or transmit data and / or other signals. Additionally, or alternatively, the processor may be configured to control one or more components of a device (e.g., console, touchscreen, personal computer, laptop, tablet, server).

[0093] In some variations, the processor 142 may be configured to access or receive data and / or other signals from one or more of workcell 110, server, control system 140, and a storage medium (e.g., memory, flash drive, memory card, database). In some variations, the processor 142 may be any suitable processing device configured to run and / or execute a set of instructions or code and may include one or more data processors, image processors, graphics processing units (GPU), physics processing units, digital signal processors (DSP), analog signal processors, mixed-signal processors, machine learning processors, deep learning processors, finite state machines (FSM), compression processors (e.g., data compression to reduce data rate and / or memory requirements), encryption processors (e.g., for secure wireless data transfer), and / or central processing units (CPU). The processor 142 may be, for example, a general-purpose processor, Field Programmable Gate Array (FPGA), an Application Specific Integrated Circuit (ASIC), a processor board, and / or the like. The processor may be configured to run and / or execute application processes and / or other modules, processes and / or functions associated with the system. The underlying device technologies may be provided in a variety of component types, such as metal-oxide semiconductor field-effect transistor (MOSFET) technologies like complementary metal- oxide semiconductor (CMOS), bipolar technologies like emitter-coupled logic (ECL), polymer technologies (e.g., silicon-conjugated polymer and metal-conjugated polymer-metal structures), mixed analog and digital, and / or the like.

[0094] The processor 142 may operate the systems / perform the methods herein using software (executed on hardware), hardware, or a combination thereof. Hardware modules may include, for example, a general -purpose processor (or microprocessor or microcontroller), a field programmable gate array (FPGA), and / or an application specific integrated circuit (ASIC). Software modules (executed on hardware) may be expressed in a variety of software languages (e.g., computer code), including structured text, typescript, C,C++, C#, Java®, Python, Ruby, Visual Basic®, and / or other object-oriented, procedural, or other programming language and development tools. Examples of computer code include, but are not limited to, micro-code or micro-instructions, machine instructions, such as produced by a compiler, code used to produce a web service, and files containing higher-level instructions that are executed by a computer using an interpreter. Additional examples of computer code include, but are not limited to, control signals, encrypted code, and compressed code.

[0095] The memory 144 of the control system 140 may be configured to store data and / or information. In some variations, the memory may include one or more of a random-access memory (RAM), static RAM (SRAM), dynamic RAM (DRAM), a memory buffer, an erasable programmable read-only memory (EPROM), an electrically erasable read-only memory (EEPROM), a read-only memory (ROM), flash memory, volatile memory, nonvolatile memory, combinations thereof, and the like. In some variations, the memory may store instructions to cause the processor to execute modules, processes, and / or functions associated with the device, such as image processing, image display, sensor data, data and / or signal transmission, data and / or signal reception, and / or communication. Some embodiments described herein may relate to a computer storage product with a non-transitory computer- readable medium (also may be referred to as a non-transitory processor-readable medium) having instructions or computer code thereon for performing various computer-implemented operations. The computer-readable medium (or processor-readable medium) is non-transitory in the sense that it does not include transitory propagating signals per se (e.g., a propagating electromagnetic wave carrying information on a transmission medium such as space or a cable). The computer code (also may be referred to as code or algorithm) may be those designed and constructed for the specific purpose or purposes. In some variations, the memory may be configured to store any received data and / or data generated by the controller and / or workcell. In some variations, the memory may be configured to store data temporarily or permanently.

[0096] The input device 148 of the control system 140 may comprise or be coupled to a display (e.g., display 150). The input device 148 may be any suitable device that is capable of receiving input from an operator via, for example, a keyboard, buttons, touch screen, and / or the like. The input device may include at least one switch configured to generate a user input. For example, an input device may include a touch surface for a user to provide input (e.g., finger contact to the touch surface) corresponding to a user input. An input device including atouch surface may be configured to detect contact and movement on the touch surface using any of a plurality of touch sensitivity technologies including capacitive, resistive, infrared, optical imaging, dispersive signal, acoustic pulse recognition, and surface acoustic wave technologies. In embodiments of an input device including at least one switch, a switch may have, for example, at least one of a button (e.g., hard key, soft key), touch surface, keyboard, analog stick (e.g., joystick), directional pad, mouse, trackball, jog dial, step switch, rocker switch, pointer device (e.g., stylus), motion sensor, image sensor, and microphone. A motion sensor may receive user movement data from an optical sensor and classify a user gesture as a user input. A microphone may receive audio data and recognize a user voice as a user input.

[0097] Graphical and / or image data may be output on the display 150 of the controller. In some variations, a display may include at least one of a light emitting diode (LED), liquid crystal display (LCD), electroluminescent display (ELD), plasma display panel (PDP), thin film transistor (TFT), organic light emitting diodes (OLED), electronic paper / e-ink display, laser display, and / or holographic display. In some variations, a GUI may be configured for designing a process and monitoring a product and may be shown on the display.

[0098] Further, in some variations, the controller may include a communication device (e.g., communication device 126) configured to communicate with another controller and one or more databases. The communication device may be configured to connect the controller to another system (e.g., Internet, remote server, database, workcell) by wired or wireless connection. In some variations, the system may be in communication with other devices via one or more wired and / or wireless networks. In some variations, the communication device may include a radiofrequency receiver, transmitter, and / or optical (e.g., infrared) receiver and transmitter configured to communicate with one or more devices and / or networks. The communication device may communicate by wires and / or wirelessly.III. Cartridge

[0099] The cartridge 160 may be configured to carry process material throughout a bioprocessing workflow. For RNA processing, the material may be genetic material, such as DNA, RNA, derivatives thereof, proteins, lipids, and / or combinations thereof (e.g., DNA template(s), unpurified mRNA, purified mRNA, unpurified lipid nanoparticles, and / or purified lipid nanoparticles).

[0100] The cartridge 160 may comprise one or more modules configured to interface and / or engage with (e.g., releasably couple to) an instrument 120 within the workcell 110. Asused herein, the term “module”, refers to a set of components (e.g., one or more components having shared and / or dissimilar functions) that are used to complete a particular processing step, such as a portion of an RNA manufacturing process. The modules may be actuated by corresponding actuating modules of a bioprocessing instrument.

[0101] Some or all of the modules may be integrated in a fixed configuration within the cartridge, though they need not be. For example, one or more of the modules may be configurable or moveable (e.g., by an operator, controller, and / or robot of the workcell) within an enclosure of the cartridge 160, permitting various formats of instruments to be assembled. That is, a cartridge may have configurable and / or moveable modules coupled by configurable fluidic, mechanical, optical, magnetic, and / or electrical connections. Similarly, in some variations, the components of various modules herein may be interspersed with each other such that each module may be defined by the set of connected components that collectively perform a predetermined function. However, these components of a module may or may not be physically grouped within the instrument.

[0102] Various materials can be used to construct the cartridge and the cartridge housing, including metal, plastic, rubber, and / or glass, or combinations thereof. The cartridge, its components, and its housing may be molded, machined, extruded, 3D printed, or any combination thereof. The cartridge may contain components that are commercially available (e.g., tubing, valves, fittings); these components may be attached or integrated with custom components or devices. The housing of the cartridge may constitute an additional layer of enclosure that further protects the sterility of the cell product. The operator may perform loading or unloading of the cartridge in an ISO8 or cleaner environment, utilizing aseptic technique to ensure that sterility of the contents of the cartridge is maintained when the cartridge 160 is opened. In some variations, the operator may perform loading or unloading of the cartridge 160 using manual aseptic connections (e.g., sterile tube welding). The robotic system may also perform sterile loading or unloading of liquids into and out of the cartridge through the use of the sterile liquid transfer instrument and sterile liquid transfer ports on the cartridge 160. The cartridge 160 may be configured for single- or multi-use.

[0103] A fluid capacity of the cartridge 160 may be determined by a fluid capacity of each module and system therein having a fluid compartment(s). In some variations, the cartridge 160 may be configured to process a bioproduct for a large group of individuals (e.g., over 10, over 50 individuals, over 100 individuals). In such variations, the cartridge may have a fluid capacity of about 5 L to about 20 L, such as about 6 L to about 17.5 L, about 7 L to about 15L, about 8 L to about 12.5 L, or about 9 L to about 10 L (including all ranges and subranges therebetween). In some variations, the cartridge 160 may be configured to process a bioproduct for a large group of individuals (e.g., over 100 individuals, over 150 individuals, over 200 individuals, over 250 individuals, over 500 individuals, etc.). In such variations, the cartridge may have a fluid capacity of about 5 L to about 20 L, such as about 6 L to about 17.5 L, about 7 L to about 15 L, about 8 L to about 12.5 L, or about 9 L to about 10 L, including all ranges and subranges therebetween (e.g., about 7 L or about equal to or less than 7 L). In some variations, the cartridge 160 may be configured to process a small volume of product, such as for one individual. In such variations, the cartridge may have a fluid capacity of about 50 mL to about 1 L, such as about 75 mL to about 750 mL, about 100 mL to about 500 mL, about 125 mL to about 400 mL, or about 150 mL to about 300 mL, about 175 mL to about 275 mL, or about 200 mL to about 250 mL, including all ranges and subranges therebetween (e.g., about 250 mL or about equal to or less than 250 mL).

[0104] The cartridge 160 herein may additionally include systems configured to monitor and / or control one or more process parameters or conditions throughout the processing. That is, the systems may be configured to perform process maintenance, while the modules may carry out specific bioprocessing like transcription, digestion, capping & tailing, particle formation, purification, and formulation (i.e., storage and concentration). For example, the systems may be configured to manage fluid transfer (including sampling for analytics), waste collection, reagent storage, and temperature control on the cartridge. In some variations, one or more systems of the cartridge may function by engaging a corresponding system of a bioprocessing instrument.

[0105] FIG. 1C is a block diagram of an illustrative variation of the cartridge 160, which may include common housing 161 to support one or more systems and one or more modules. Systems of the cartridge 160 may include a fluid transfer system (FTS) 162, a storage system 166, a waste system 167, and a temperature control system (TCS) 168, and an analytical system 169. In some variations, components of these systems may be shared, and / or two or more of these systems may work together. For example, as described below, the TCS 168 may be configured to precisely control a temperature of one or more fluid compartments of the storage system 166. As described below, one or more cartridge systems may include fluid compartments. In some variations, the fluid compartments may be reconfigurable in size (e.g., fluid capacity) and / or number to provide workflow flexibility and process scalability.1. Fluid Transfer System (FTS)

[0106] The FTS 162 may include a fluidic bus configured to couple to one or more of the cartridge systems and modules (e.g., all of them) via fluid conduit(s). In some variations, the fluidic bus may be located (e.g., centrally) within a housing of the cartridge 160. The fluidic bus may thus fluidically connect the cartridge systems and modules to enable process flow. The FTS 162 may also include one or more interfaces for coupling to a pump actuator of an instrument. For example, the pump actuator may be configured to engage one or more portions of fluidic conduits (e.g., tubing) of the fluidic bus to drive fluid flow throughout the cartridge 160. Further, the FTS 162 may include one or more ports configured to enable sampling of fluid from within the cartridge 160. In particular, the port(s) may facilitate sterile transfer of samples for analysis (e.g., by instruments of the AIS and / or offline instruments) throughout a bioprocessing workflow such that sterility of the cartridge and product therein maybe maintained throughout.2. Storage System

[0107] The storage system 166 may be configured to store reagents for use during a bioprocessing workflow. In some variations, reagents may be preloaded into the storage system. Additionally, or alternatively, reagents may be added or removed therefrom at any point during the workflow. The cartridge 160 may be configured to engage the RMS 180 to have such reagents added and / or removed.

[0108] The storage system 166 may include one or more fluid compartments, such as a plurality thereof, for storing different reagents and for providing varying storage conditions for the reagents. In some variations, the storage system 166 may comprise 1 to 20 fluid compartments, such as 2 to 15, 3 to 14, 4 to 13, 5 to 12, 6 to 11, 7 to 10, or 8 to 9 fluid compartments, including all ranges and subranges therebetween (e.g., 5, 6, 7, 8, 9, or 10 fluid compartments). A plurality of fluid compartments may include fluid compartments of various sizes. Each fluid compartment may have a fluid capacity of about 50 mL to about 500 mL, such as about 75 mL to about 450 mL, about 100 mL to about 400 mL, about 125 mL to about 350 mL, about 150 mL to about 300 mL, about 175 mL to about 250 mL, including all ranges and subranges therein, such as about 200 mL. A total fluid capacity of the storage system 166 may be about 1 L to about 5 L, such as about 1.25 L to about 3 L, about 1.5 L to about 2.75 L, about 1.75 L to about 2.5 L, or about 2 L to about 2.25 L.

[0109] Each fluid compartment of the storage system 166 may be temperature-controlled by the TCS of the instrument 120. Each fluid compartment may be adjacent to or in contact with one or more (e.g., at least one) thermal element of the TCS. In some variations, temperatures within the storage system 166 may be maintained at about 2 deg C to about 8 deg C, such as at about 4 deg C. Accordingly, the storage system may provide a suitable environment for one or more reagent types, including enzymes, buffers, and / or lipids used throughout RNA processing.3. Waste System

[0110] The waste system 167 may be configured to collect, and in some variations, segregate wastes formed within the cartridge 160 during bioprocessing. The waste system 167 may include one or more fluid compartments, such as a plurality thereof, for storing wastes. In some variations, the fluid compartments may comprise a, or at least one, first fluid compartment configured to store aqueous waste (e.g., from the TFFM and / or CM), and a, or at least one, second fluid compartment configured to store organic waste (e.g., from the PFM). The first and second fluid compartments may be a same or different size. For example, a fluid capacity of one or both of the first and second fluid compartments may be about 0.25 L to about 2 L, such as about 0.5 L to about 1.75 L, about 0.75 L to about 1.5 L, about 0.8 L to about 1.25 L, about 0.9 L to about 1.1 L, including all ranges and subranges therebetween (e.g., about 1 L). A total fluid capacity of the waste system 167 may be about 1 L to about 5 L, such as about 1.25 L to about 3 L, about 1.5 L to about 2.75 L, about 1,75 L to about 2.5 L, or about 2 L to about 2.25 L.

[0111] In some variations, each fluid compartment of the waste system 167 may be temperature-controlled by the TCS of the instrument 120. For example, fluid compartment may be adjacent to or in contact with one or more (e.g., at least one) thermal element of the TCS. In some variations, temperatures within the waste system 167 may be maintained at about 2 deg C to about 8 deg C, such as at about 4 deg C. Further, in some variations, one or more sensors (e.g., bubble sensor(s), flow sensor(s), pressure sensor(s). and / or camera(s)) of the cartridge 160 and / or an interfacing instrument 120 may be configured to detect a fluid level and / or flow rate within the waste system 167 (e.g., within one or more fluid compartments thereof) to prevent overflow therein. When the waste system 167 is at capacity or about full, the cartridge 160 may be transferred to the RMS 180 or WMS 114 of the workcell so that the waste be removed.4. Temperature Control System (TCS)

[0112] The TCS 168 may include one or more on-board temperature sensors configured to detect (e.g., continuously) temperatures within one or more modules of the cartridge 160. The temperature sensors may provide continuous feedback to the control system 140, which may determine whether the measurements are acceptable or out of range. When out of range, the control system 140 may adjust a temperature applied by the thermal elements (of the instrument TCS). In some variations, each cartridge module may include one or more (e.g. at least one) temperature sensor of the TCS 168. In some variations, each cartridge module requiring temperature control may include one or more (e.g. at least one) temperature sensor of the TCS 168.5. Analytical System

[0113] The cartridge 160 may include an integrated analytical system 169 comprising onboard sensor(s) and / or analytical tool(s) for monitoring a bioprocessing workflow in realtime. In some variations, one or more components of the analytical system 169 may be communicab ly coupled to the control system 140 such that continuous feedback on the workflow may be analyzed by the control system 140. Tire control system 140 may use this information to inform subsequent operations of the workflow (e.g., to modify or verify such subsequent operations).

[0114] In some variations, the analytical system 169 may include one or more sensors for continuous or periodic detection of process and / or product parameters throughout a bioprocessing workflow. The sensor(s) may include one or more pH sensor(s), conductivity sensor(s), temperature sensor(s), pressure sensor(s), bubble sensor(s), and / or camera(s). The pH, conductivity, and temperature sensor(s) may be configured to provide real-time feedback on sample parameters (pH, conductivity, temperature), and the pH sensor(s) may be configured to provide real-time feedback on process parameters like fluid flow rate throughout the FTS 162, pressure differential of the TFFM, and the like. The bubble sensor(s) and / or camera(s) may provide real-time feedback on cartridge parameters like a fluid level within a module.

[0115] Additionally, or alternatively, the analytical system 169 may include one or more liquid chromatographic columns for determining purity percent, yield, and / or yield percent of RNA material (e.g., crude RNA, DNase-treated mRNA, purified mRNA, capped and tailedmRNA, purified capped and tailed mRNA, mRNA-lipid product, and / or purified and concentrated mRNA-lipid product). Additionally, or alternatively, the analytical system 169 may include one or more UV-Vis spectrophotometers for determining mRNA concentration and purity of the RNA material (e.g., one or more of the crude RNA, the DNase-treated mRNA, the purified mRNA, the capped and tailed mRNA, and the purified capped and tailed mRNA). Additionally, or alternatively, the analytical system 169 includes one or more Raman spectrophotometers for detecting contaminants in any intermediate or final product throughout a bioprocessing workflow.

[0116] The cartridge modules described below may each be configured to perform a step of a bioprocessing workflow, such as of an RNA processing workflow. Such a workflow may include one or more of each of a transcription step, a digestion step, a capping / tailing step, a purification step, a particle formation step, and a concentration step. In some variations, one or more of the modules herein may share some or all of a set of components. The cartridge 160 may be configured with all or fewer than all of the following modules.6. Transcription Module

[0117] The transcription module 170 may comprise an in vitro transcription (IVT) module. Material entered into the transcription module 170 may include buffer, transcription enzymes, and DNA templates. The transcription module 170 may comprise at least one fluid compartment, or mixing chamber, for supporting reactions. The fluid compartment may include a magnetic stirrer or paddle that may be actuated by the instrument 120 to provide homogeneous reaction conditions within the fluid compartment. The mixing chamber may have a fluid capacity of between about 100 mL and about 300 mL, such as about 200 mL to about 250 mL (e.g., about 200 mL or about 250 mL). The TCS may engage with the transcription module 170 to provide a controlled temperature therein of about 30 deg C to about 50 deg C, such as about 35 deg C to about 45 deg C, or about 37 deg C to about 42 deg C (e.g., about 37 deg C). One or more sensors on the cartridge and / or instrument, such as temperature and / or pH sensors, may also help maintain reaction conditions within the transcription module 170 by providing continuous feedback on reaction conditions.7. Digestion Module

[0118] The digestion module 171 may be a DNase digestion module for digesting portions of untranscribed DNA template following transcription by the transcription module 170.Accordingly, the output from the transcription module 170 may enter into the digestion module 171 for processing. In some variations, the digestion module 171 and the transcription module 170 may share one or more components, such as the same fluid compartment / mixing chamber. Like the transcription module 170, the (same or different) mixing chamber of the digestion module 171 may have a fluid capacity of between about 100 mL and about 300 mL, such as about 200 mL to about 250 mL (e.g., about 200 mL, about 250 mL, or about 300 mL). The TCS may engage with the digestion module 171 to provide a controlled temperature therein of about 30 deg C to about 50 deg C, such as about 35 deg C to about 45 deg C, or about 37 deg C to about 42 deg C (e.g., about 37 deg C). One or more sensors on the cartridge and / or instrument, such as temperature and / or pH sensors, may also help maintain reaction conditions within the digestion module 171 by providing continuous feedback on reaction conditions.8. Purification Module

[0119] The purification module 172 may include one or more sub-modules including one or more of a tangential flow filtration module (TFFM), a magnetic selection module (MSM), and a chromatography module. The purification module 172 may include any combination of these modules for purifying intermediate and / or final RNA products.

[0120] The TFFM may be configured to concentrate and purify RNA, exchange buffers, and remove impurities in the sample material. To do so, the TFFM may include a tangential flow filtration (TFF) system including feed, retentate, and permeate reservoirs, as well as a membrane for flowing the sample across. The reservoirs may each have a fluid capacity of about 200 mL to about 550 mL, such as about 250 mL to about 500 mL. In some variations, one or more of the reservoirs may have a unique fluid capacity. For example, in some variations, the feed reservoir and the permeate reservoir may have a fluid capacity of about 500 mL, and the permeate reservoir may have a fluid capacity of about 250 mL. In some variations, the membrane may have an area of about 0.01 m2 to about 0.05 m2, and pore size that is about equal to or less than a size of the RNA material.

[0121] The TCS may engage with the TFFM to provide a controlled temperature therein of about 4 deg C to about 25 deg C. One or more sensors on the cartridge and / or instrument, such as one or more pressure sensors, may also help maintain reaction conditions within the TFFM.

[0122] The MSM may comprise one or more flowcells for flowing a sample therethrough (via fluid conduits of the fluidic bus). The sample may be mixed with magnetic beads such that the material therein may be sorted via magnetic separation. The flowcell may have a volume of about 10 mL to about 100 mL.

[0123] The chromatography module may be configured to purify the product by removing impurities and truncated sequences. The chromatography module may include one or more vessels for loading, chromatography, and elution of an RNA product. A load preparation fluid compartment may have a fluid capacity of about 700 mL to about 800 mL, such as about 750 mL. A chromatography column may have a fluid capacity of about 25 mL to about 100 mL, such as about 40 mL, about 50 mL, or about 60 mL. An elution collection fluid compartment may have a fluid capacity of about 200 mL to about 300 mL, such as about 250 mL.9. Capping and Tailing Module (C&TM)

[0124] The C&TM 173 may be configured to synthesize mRNA from pre-mRNA by providing a cap and tail (e.g., 5' cap and 3' poly(A) tail) to the pre-mRNA. In some variations, the C&TM 173, the digestion module 171, and / or the transcription module 170 may share one or more components, such as the same fluid compartment / mixing chamber. In some variations, the cartridge 160 may include a single set of components (e.g., mixing chamber, magnetic stirrer, sensors, etc.) for using each of the C&TM 173, the digestion module 171, and the transcription module 170. Together, these modules may form a more general enzymatic reaction module, or ERM.

[0125] Like the digestion module 171 , a mixing chamber of the C&TM 173 may have a fluid capacity of between about 100 mL and about 300 mL, such as about 200 mL to about 250 mL (e.g., about 200 mL, about 250 mL, or about 300 mL). The TCS may engage with the digestion module 171 to provide a controlled temperature therein of about 30 deg C to about 50 deg C, such as about 35 deg C to about 45 deg C, or about 37 deg C to about 42 deg C (e.g., about 37 deg C). One or more sensors on the cartridge and / or instalment, such as temperature and / or pH sensors, may also help maintain reaction conditions within the digestion module 171 by providing continuous feedback on reaction conditions.10. Particle Formation Module (PFM)

[0126] The PFM 174 may be an RNA-loaded lipid nanoparticle (LNP) module. In particular, the PFM 174 may be configured to encapsulate RNA in lipid (nano) particles sothat the RNA may be capable of entering the cytosol of a patient. To support this particle formulation, the PFM 174 may include at least one fluid compartment, or mixing chamber, configured to engage thermal element(s) and temperature sensor(s) of the TCS. In some variations, the PFM 174 may include a mixing chamber, a mixer (e.g., microfluidic mixer, such as one of standard herringbone design), and a stabilization chamber. The mixing chamber may have a fluid capacity of between about 50 mL and about 500 mL, such as about 100 mL. The stabilization chamber may also have a fluid capacity of between about 50 mL and about 500 mL, such as about 400 mL. The TCS may engage the mixing chamber to maintain a temperature of about 20 deg C to about 40 deg C (e.g., 30 deg C) during mixing. After being mixed, the product may be rapidly cooled in the stabilization chamber (via the TCS) to a temperature of about 2 deg C to about 8 deg C (e.g., about 4 deg C). The mixer may include two inlet ports fluidically coupled to the fluidic bus 162. The mixer may receive the RNA (mRNA in aqueous buffer) via a first of the two inlets, and the lipids (e.g., LNPs in ethanol) through a second of the two inlets. From the outlet of the mixer, the fluidic bus 162 may direct the RNA-lipid product back to the stabilization chamber for cooling.

[0127] In some variations, the PFM 174 may include one or more on-cartridge pressure sensors for providing flow verification and clog detection. Additionally, real-time feedback systems described herein, like in-line DLS, may be used with the PFM 174 to control flow rate of the product therethrough.11. Final Formulation Module (FFM)

[0128] The FFM 175 may be configured to store a final mRNA product. Material entered into the FFM 175 may include an RNA-lipid suspension (e.g., mRNA-LNP suspension) The FFM 175 may comprise at least one fluid compartment for storing the RNA product. The fluid compartment may have a fluid capacity of between about 100 mL and about 700 mL, such as about 500 mL. The TCS may engage the FFM 175 to provide a controlled temperature therein of about 2 deg C to about 8 deg C, such as about 4 deg C. In some variations, the FFM 175 may be configured to filter and / or concentrate the RNA-lipid product. For example, the FFM 175 may include a membrane for sterile filtration of the product. In some variations, the product may be transferred out of the workcell 110 within the FFM 175 of the cartridge 160 when the bioprocessing protocol is complete.

[0129] In some variations, the cartridge 160 may include one or more additional modules, such as a final formulation module configured to repurify and concentrate an RNA-lipid product.

[0130] FIG. 3 is a schematic diagram of an exemplary' variation of a cartridge 300 for automated bioprocessing. As shown, the cartridge 300 may include an FTS having pump(s) 304, fluidic bus 306 and port(s) 308 for transferring fluid among the cartridge modules and for providing sterile transfer of samples for analysis. The fluidic bus 306 may be centrally positioned within the cartridge 300 such that each cartridge module may couple thereto via one or more fluid conduits. The cartridge modules may include, on a first side of the fluidic bus 306, a TM 310, a digestion module 312, and purification module 314 including one or more of a TFFM, MSM, and chromatography module. On a second, opposite side of the fluidic bus 306, PFM 316, FFM 318, and reagent and waste systems 320, 322, respectively, may be positioned.

[0131] In summary, the automated bioprocessing systems described herein provide a comprehensive, integrated platform for end-to-end manufacturing of nucleic acid therapeutics, particularly mRNA products. The workcell, with its controlled environment and integrated subsystems (including the MHS, RMS, and AIS), may work in concert with the modular cartridge to enable fully automated, sterile processing from DNA template to final formulated product. The integrated modules of the cartridge- spanning transcription, digestion, purification, particle formation, and final formulation - may interface with corresponding actuating modules of the bioprocessing instruments, while the control system may orchestrate the entire workflow, monitor process parameters in real-time, and enable responsive process control. This architecture may support both personalized medicine applications requiring small-batch production and larger-scale manufacturing needs, with the flexibility to process multiple cartridges in parallel for increased throughput. The following section describes methods for utilizing these systems and devices to execute automated bioprocessing workflows, including specific protocols for mRNA manufacturing and adaptive process control strategies that leverage the real-time monitoring and analysis capabilities of the integrated platform.Methods for Automated Bioprocessing

[0132] Described herein are also methods for bioprocessing, such as methods for processing RNA (e.g., mRNA therapeutics). The methods may be for use with or executed bythe automated bioprocessing systems and devices, such as with or by the system 100 of FIGS. 1A-1E and variations thereof. While the methods herein may be described as having particular steps with a particular order, it should be understood that, in some variations: every step of the method may not be performed (e.g., may be optional), and / or the steps of the method may be performed in a different order, and / or two or more steps of the method may be performed simultaneously (or overlap), and / or one or more additional steps may be performed.

[0133] FIG. 4 provides a flowchart of an illustrative variation of a method for automated bioprocessing using the systems and devices herein. The method 400 may first include loading 402 a cartridge with a product into a workcell. The cartridge may be loaded into a feedthrough of the workcell. Optionally, the loading 402 may include sterilizing the cartridge within the feedthrough, such as via hydrogen peroxide or another decontaminant. In some variations, the sterilization may involve exposing the cartridge to vaporized hydrogen peroxide (VHP) for a duration sufficient to achieve a target sterility assurance level (SAL). For example, the feedthrough may also include UV-C sterilization capabilities for additional decontamination. During loading, sensors within the feedthrough may scan identification markers (e.g., RFID tags or barcodes) on the cartridge to verify the cartridge type, batch information, and intended processing protocol.

[0134] Next, the cartridge may be transferred 404 to an instrument of the workcell, such as to a bioprocessing instrument. The transferring 404 may be performed by a robot within the workcell. In some variations, the robot may utilize precision end effectors to securely grasp the cartridge and navigate along predetermined paths (e.g., via ceiling or floor-mounted rails) to avoid contamination. For example, the transfer may include alignment procedures to ensure proper mechanical, electrical, fluidic, and thermal interfaces between the cartridge and instrument. The robot may verify successful docking through feedback sensors before releasing the cartridge.

[0135] The method 400 may then include performing 406 a bioprocessing operation on the product of the cartridge. The bioprocessing operation may include a portion of a biotherapeutic manufacturing workflow, such as a portion of an mRNA manufacturing protocol. For example, the operation may include transcription, digestion, capping, tailing, purification, concentration, particle formation, and storage. In some variations, during transcription, the instrument may maintain the transcription module at about 30°C to about 50°C (e.g., about 37°C) while actuating magnetic stirrers at about 150 RPM to about 200RPM to ensure homogeneous mixing of DNA template, RNA polymerase, and reaction buffer. For purification operations, the instrument may control tangential flow filtration parameters to optimize RNA recovery while removing impurities. In some variations, more than one bioprocessing operation may be performed during the step 406. For instance, digestion and capping may occur simultaneously within the same reaction chamber, with DNase enzymes removing residual DNA template while capping enzymes add the 5' cap structure to the nascent mRNA. Tire performing 406 may be achieved via the instrument (e.g., one or more actuating modules thereof) actuating the cartridge (e.g., one or more modules thereof). This step 406, or a variation thereof (e.g., performing a modified or different bioprocessing operation), may be repeated any number of times until the workflow is complete.

[0136] Next, the method 400 may include performing 408 an analytical operation on the product. The analytical operation may include using one or more sensors and / or analytical instruments or tools to detect a product parameter (e.g., % purity, % yield, yield, concentration, and / or the like) or condition (e.g., molecule type, particle type). For example, UV-Vis spectrophotometry may be used to determine RNA concentration and purity. In some variations, HPLC analysis may quantify percent purity by separating full-length mRNA from truncated sequences and other impurities. For lipid nanoparticle characterization, dynamic light scattering (DLS) may be employed to measure particle size distribution and polydispersity.

[0137] The sensor(s) and / or analytical instrument(s) may be positioned on the cartridge, the instrument, and / or elsewhere in the workcell, such as at the AIS 190. hi some variations, the step 408 may include transferring and / or coupling an analytical instrument to the cartridge to retrieve a sample of the product therefrom. Sample volumes may range from about 0 iiL to about 300 pL depending on the analytical technique, and may be withdrawn through dedicated sampling ports that maintain sterility through septum or valve mechanisms. In some variations, the step 408 may include transferring the cartridge from the instrument to elsewhere in the workcell, such as the RMS 180 or AIS 190, so that the product may be sampled and analyzed. In some variations, a robot of the workcell may perform such transferring (of the cartridge and / or the analytical tool). The analysis may determine whether the bioprocessing workflow may proceed as planned, or whether the workflow may be modified (as discussed in more detail below with reference to FIG. 5). In some variations, real-time data from the analysis may be transmitted to the control system 140, whichcompares measured values against predefined quality attributes and acceptance criteria. Like the step 406, the step 408, or a variation thereof (e.g., such as performing a modified or different analytical operation), may be repeated any number of times until the workflow is complete.

[0138] Once the workflow is complete, the method 400 may include unloading 410 the cartridge (and thus the product therein) from the workcell. In some variations, the workflow may be complete when, after a final bioprocessing operation in the protocol is performed, an analysis of the product results in confirmation that the product has achieved one or more standards (e.g., of purity, yield, concentration, and / or the like). For example, final product specifications may require the product to meet predetermined thresholds for parameters such as intact mRNA percentage, encapsulation efficiency for lipid nanoparticles, endotoxin levels, and residual DNA content. The unloading 410 may include transferring the cartridge (e.g., via the robot) to a feedthrough, which may be a same or different feedthrough through which the cartridge was loaded into the workcell. In some variations, the unloading 410 may include sterilizing the cartridge within the feedthrough. This post-processing sterilization may prevent any potential contamination from exiting the controlled environment and may utilize UV-C exposure or chemical sterilants appropriate for the filled product. Finally, the method 400 may include generating 412 an electronic batch record for the product. In some variations, the electronic batch record may compile all process parameters, analytical results, deviations, and operator actions throughout the workflow, and may be formatted to meet regulatory compliance requirements

[0139] Turning to FIG. 5, an illustrative variation of another method for automated bioprocessing using the systems and devices herein is shown. Like the method 400, the method 500 may include first loading 502 a cartridge cartying a product into a bioprocessing workcell, and performing 504 at least a portion of a bioprocessing workflow on the product. In some variations, at least a portion of the workflow may include one or more partially or frilly completed bioprocessing operations, such as the mRNA manufacturing operations discussed herein throughout, of the workflow. For instance, the workflow portion may encompass the complete enzymatic synthesis phase (transcription, digestion, and capping / tailing) before proceeding to downstream purification, or may include only the transcription step before pausing for quality assessment.

[0140] Next, the method 500 may include analyzing 506 the product. The analyzing 506 may involve using one or more sensors and / or analytical instruments or tools to measure aproduct parameter (e.g., % purity, % yield, yield, concentration, and / or the like) or detect a product condition (e.g., molecule type, particle type). In some variations, specific analytical methods may include reverse-phase HPLC for separating capped from uncapped mRNA species, gel electrophoresis for assessing RNA integrity, quantitative PCR for detecting residual DNA template, and appropriate assays for endotoxin testing. The sensor(s) and / or analytical instrument(s) may be positioned on the cartridge, the instrument, and / or elsewhere in the workcell, such as at the AIS 190. In some variations, the step 408 may include transferring and / or coupling an analytical instrument to the cartridge to retrieve a sample of the product therefrom. In some variations, the analyzing 506 may include transferring the cartridge from the instrument to elsewhere in the workcell, such as the RMS 180 or AIS 190, so that the product may be sampled and analyzed. In some variations, a robot of the workcell may perform such transferring (of the cartridge and / or the analytical tool).

[0141] hi some variations, the analyzing 506 may include determining whether the detected parameter or condition meets a standard, such as a threshold value (or is within an acceptable range of values), such as by comparing the parameter or condition to the threshold. For example, the control system may compare measured yield values against predetermined thresholds (e.g., about 50% to about 100%, such as about 75% to about 80%) and flag any values falling outside the acceptable range as requiring process modification. In some variations, the analyzing 506 may be repeated any number of times using one or more same or different sensor(s) and / or analytical instrument(s). Multiple analytical techniques may be employed in parallel or series to provide orthogonal confirmation of product quality.

[0142] Next, the method 500 may include modifying 508 (e.g., via a control system) the bioprocessing workflow when the analyzed product does not meet a standard. In some variations, the modifying may include delaying one or more operations, replacing one or more operations, repeating one or more operations, canceling one or more operations, and / or adjusting parameters for one or more operations of the protocol. Examples of specific modifications may include: extending enzyme reaction times if yield is suboptimal, adding additional purification cycles if purity falls below threshold values, adjusting buffer conditions to improve enzyme activity, modifying mixing speeds within the range of about 0 RPM to about 500 RPM to enhance mass transfer, implementing buffer exchange steps if conductivity exceeds specifications, or adjusting formulation parameters to improve product characteristics. In some variations, the control system 140 may automatically implementthese modifications based on predefined decision trees or algorithms utilizing historical process data.

[0143] An exemplary method 600 for manufacturing mRNA is detailed in FIG. 6. The method 600 may be performed with the systems and devices herein, such as with the cartridge 160, workcell 110, and controller 140 of the system 100 of FIGS. 1 A- IE. At step 602, transcription, DNA template is mixed with RNA polymerase, buffer, RNase inhibitor, and pyrophosphatase. At step 604, enzymatic addition, the crude mRNA output (e.g., pre- rnRNA) is mixed with buffer and one or more of DNase, capping enzyme(s) (e.g., RNA triphosphatase, guanylyltransferase, and methyltransferase), tailing enzyme(s) (e.g., poly(a) polymerase). Accordingly, the enzymatic addition at step 604 may capture one or both of digestion and capping & tailing (which may be carried out simultaneously). In some variations, the enzymatic addition (e.g., digestion and / or capping & tailing) may be performed by controlling a mixing speed and temperature of the solution. At step 606, the mRNA product from step 604 is mixed with diafiltration buffer and stabilizers (e.g., EDTA) to undergo purification and concentration via tangential flow filtration. At step 607, wastes collected during steps 602-606 are collected. Next, at step 610, the mRNA product from step 608 is mixed with functionalized beads (e.g., magnetic carrier material) that attach to contaminants within the product so that the product may undergo purification via magnetic separation. As shown in FIG. 6, in some variations, both of steps 608 and 610 may be performed. In alterative variations, only one of steps 608 and 610 may be performed. At step 612, the mRNA product from step 608 and / or step 610 may be mixed with solvent, lipids (e.g., LNPs), and buffer to complete particle formation of mRNA-encapsulated lipids. In some variations, the mixing may be achieved by controlling a flow rate of the solution through a mixer, such as a micro fluidic mixer. At step 614, the mRNA-lipid product is mixed with buffer for purification. In some variations, the purification may involve sterile filtration. Following step 614, the finished product may be stored or shipped to a patient / institution for use.

[0144] Throughout this application, the term “about” is used to indicate that a value includes the inherent variation of error for the device or the method being employed to determine the value, or the variation that exists among the samples being measured. Unless otherwise stated or otherwise evident from the context, the term “about” means within 10% above or below the reported numerical value (except where such number would exceed 100% of a possible value or go below 0%). When used in conjunction with a range or series ofvalues, the term “about” applies to the endpoints of the range or each of the values enumerated in the series, unless otherwise indicated. As used in this application, the terms “about” and “approximately” are used as equivalents.

[0145] Additionally, it should be appreciated that ranges disclosed herein may be exemplary, and include all ranges and subranges therein.

[0146] While certain variations are described and illustrated herein, those of ordinary skill in the art will readily envision a variety of other means and / or structures for performing the function and / or obtaining the results and / or one or more of the advantages described herein, and each of such variations and / or modifications is deemed to be within the scope of the inventive variations described herein. More generally, those skilled in the art will readily appreciate that all parameters, dimensions, materials, and configurations described herein are meant to be exemplary and that the actual parameters, dimensions, materials, and / or configurations will depend upon the specific application or applications for which the inventive teachings is / are used. Those skilled in the art will recognize or be able to ascertain using no more than routine experimentation, many equivalents to the specific inventive variations described herein. It is, therefore, to be understood that the foregoing variations are presented by way of example only and that, within the scope of the appended claims and equivalents thereto; inventive variations may be practiced otherwise than as specifically described and claimed. Inventive variations of the present disclosure are directed to each individual feature and / or method described herein. In addition, any combination of two or more such features and / or methods, if such features and / or methods are not mutually inconsistent, is included within the inventive scope of the present disclosure.

Claims

CLAIMSWhat is claimed is:

1. An automated system for bioprocessing, comprising: a workcell comprising a feedthrough and at least one instrument for processing nucleic acid material; a cartridge for processing the nucleic acid material, wherein the cartridge is configured to interface with the at least one instrument, wherein the cartridge comprises: a transcription module and a purification module, wherein a temperature within at least a portion of each of the transcription and purification modules is maintained by a temperature control system of the at least one instrument; and a pump for transferring the nucleic acid material through the transcription and purification modules; a robotic arm for transferring the cartridge between the feedthrough and the at least one instrument.

2. The system of claim 1, wherein the workcell comprises an enclosure configured to maintain a sterile environment from a beginning to an end of the bioprocessing.

3. The system of claim 2, wherein the sterile environment comprises an ISO8 cleanroom.

4. The system of claim 1, wherein the cartridge comprises a first cartridge, the system further comprising a second cartridge for processing nucleic acid material, wherein the system is configured to process the nucleic acid material in each of the first and second cartridges in parallel.

5. The system of claim 1, wherein the workcell further comprises a reagent management system comprising a storage compartment for storing reagents.

6. The system of claim 5, wherein a temperature of the storage compartment is configured to be maintained within a desired range.

7. The system of claim 5, wherein the reagent management system comprises an interior zone that is separate from an interior zone of the workcell.

8. The system of claim 5, wherein the feedthrough comprises a first feedthrough, and wherein the robotic arm is configured to transfer the cartridge to and from the reagent management system via a second, different feedthrough.

9. The system of claim 1 further comprising an analytical instrument system comprising one or more analytical instruments for determining one or more parameters of an RNA product of the system.

10. The system of claim 9, wherein the one or more parameters comprise one or more of a percent purity, a percent yield, and a yield of the RNA product.

11. The system of claim 1 further comprising a controller communicably coupled to the at least one instrument and the robotic ann.

12. The system of claim 11, wherein the controller is configured to generate an electronic batch record for an RNA product of the system.

13. The system of claim 1, wherein the nucleic acid material comprises material to form mRNA.

14. An automated system for bioprocessing, comprising: a workcell comprising at least one instrument for processing nucleic acid material, a feedthrough for introducing materials into the workcell, and a robotic arm for transferring materials within the workcell; and a cartridge for processing tire nucleic acid material, wherein the cartridge is configured to interface with the at least one instrument, the cartridge comprising a transcription module, a purification module, an integrated pump for transferring the nucleic acid material between and through the transcription and purification modules, wherein at least one instrument includes a temperature control system configured to maintain a temperature within at least a portion of each of the transcription and purification modules of the cartridge, and one or more mechanical interfaces for engaging with the cartridge, andwherein the robotic arm is configured to transfer the cartridge between the feedthrough and at least one instrument.

15. A cartridge for automated bioprocessing, comprising: a plurality of modules, each module configured to interface with at least one instrument of a workcell to process nucleic acid material, wherein the plurality of modules comprises: a transcription module; a purification module; and a particle formation module; a fluidic bus for transferring fluid through the plurality of modules; and a tube configured to engage a pump for driving fluid flow through the fluidic bus.

16. The cartridge of claim 15, wherein a temperature of one or more of the transcription, purification, and particle formation modules is controlled by the at least one instrument.

17. The cartridge of claim 15 further comprising a digestion module, wherein the transcription and digestion modules comprise one or more shared components.

18. The cartridge of claim 17, wherein the one or more shared components comprise a fluid compartment.

19. Tire cartridge of claim 15, wherein the purification module comprises one or more of a tangential flow filtration module, a magnetic separation module, and a chromatography module.

20. The cartridge of claim 15 further comprising a final formation module configured to store a final product of the nucleic acid material, wherein the final product comprises RNA.

21. The cartridge of claim 15 further comprising a waste module comprising a first fluid compartment for storing aqueous waste and a second fluid compartment for storing organic waste.

22. The cartridge of claim 15 further comprising a storage module comprising a fluid compartment for storing a reagent, wherein the fluid compartment is configured to interface with a thermal element of the at least one instrument, and wherein the thermal element is configured to control a temperature of the reagent within the fluid compartment.

23. The cartridge of claim 15, wherein the cartridge comprises a housing comprising one or more ports for transferring samples of nucleic acid material to an analytical instalment during processing.

24. The cartridge of claim 15, wherein the cartridge is a single-use cartridge.

25. A cartridge for automated bioprocessing, comprising: an enclosure comprising a plurality of processing modules, wherein the plurality of modules is configured to perfonn a series of bioprocessing operations; a fluidic bus fluidically coupling the plurality of modules; one or more ports for fluid transfer; a plurality of reservoirs for storing reagents, buffers, waste; one or more external interfaces configured to engage with a bioprocessing instalment, wherein the one or more external interfaces are configured to facilitate one or both of energy and material transfer between the cartridge and the instalment; and one or more sensors for monitoring process parameters, wherein the cartridge is configured to produce nucleic acid-based products.

26. An instrument for automated bioprocessing, comprising: a receiving bay configured to receive and support a cartridge carrying material for RNA processing; a plurality of actuating modules, each module configured to drive one or more modules of the cartridge to process the material, wherein the plurality of actuating modules comprises a transcription actuation module configured to actuate a transcription reaction within the cartridge; and a temperature control system configured to maintain a temperature within the one or more modules of the cartridge, wherein the instrument is housed within an enclosed workcell.

27. The instrument of claim 26 further comprising: a first thermal control element configured to control a first temperature of a transcription module of the one or more modules of the cartridge; and a second thermal control element configured to control a second temperature of a purification module of the one or more modules of the cartridge.

28. The instrument of claim 27, wherein the first thermal control element is a component of the transcription actuation module.

29. The instrument of claim 27 further comprising a purification actuation module, wherein the second thermal control element is a component of the purification actuation module.

30. The instrument of claim 26 further comprising one or more sensors configured to monitor a parameter of the cartridge throughout the RNA processing.

31. The instrument of claim 30, wherein the one or more sensors comprise one or more bubble sensors, and wherein the parameter comprises a fluid level within the one or more modules of the cartridge.

32. The instrument of claim 30, wherein the one or more sensors are configured to transmit real-time measurements of the parameter to a controller of the workcell.

33. The instrument of claim 26 further comprising a pump configured to engage one or more fluid conduits of the cartridge to pump fluid through the cartridge.

34. An instrument for automated bioprocessing, comprising: a housing structure configured to maintain controlled environmental conditions; a receiving bay configured to receive and interface with a self-contained bioprocessing cartridge; a plurality of actuating modules configured to engage with corresponding modules of the cartridge to perform an automated bioprocessing workflow;a temperature control system configured to maintain a plurality of discrete thermal zones of the cartridge; a fluid transfer system for transferring materials to and from said cartridge; and one or more sensors for real-time process monitoring and quality assessment, wherein the instrument is configured to perform the automated bioprocessing workflow in conjunction with the cartridge to produce a finished nucleic acid therapeutic product.

35. A method for automated bioprocessing, comprising: providing a workcell comprising: one or more instalments; a feedthrough; and a robotic arm; via the robotic arm: transferring a first cartridge from the feedthrough to a first of the one or more instruments, wherein the first cartridge is configured to process a first nucleic acid material; and transferring a second cartridge from the feedthrough to a second of the one or more instruments, wherein the second cartridge configured to process a second nucleic acid material, and wherein both of the first and second cartridges comprise at least a transcription module and a purification module; and processing the first and second nucleic acid material in parallel.

36. The method of claim 35, wherein processing the first and second nucleic acid materials comprises performing a series of bioprocessing steps on each of the first and second nucleic acid materials.

37. The method of claim 36, wherein the sequence of bioprocessing steps comprises a transcription step, a digestion step, a purification step, a particle formation step, and a final formulation step.

38. The method of claim 35, wherein processing the first and second nucleic acid materials comprises performing one or more analysis steps on each of the first and second nucleic acid materials.

39. The method of claim 38, wherein performing the one or more analysis steps comprises determining one or more of a yield, a percent yield, and a percent purity of the first and second nucleic acid materials.

40. The method of claim 35 further comprising, prior to transferring the first and second cartridges to the first and second instruments, respectively, sterilizing the first and second cartridges within the feedthrough.

41. The method of claim 35 further comprising transferring the first and second cartridges from the first instrument and the second instrument, respectively, to the feedthrough after the first and second nucleic acid materials are processed.

42. A method for automated bioprocessing, comprising: transferring a cartridge from a feedthrough of a workcell to an instrument of the workcell via a robotic arm, wherein the cartridge comprises a transcription module and a purification module, and wherein the instrument is configured to perform one or more processing steps on material within the cartridge; actuating the transcription module to enable a transcription reaction therein; determining a yield of the transcription reaction; and when the yield is about equal to or greater than a threshold, transferring the material from the transcription module to the purification module.

43. The method of claim 42, wherein determining the yield comprises transferring a sample of the material to an analytical tool.

44. The method of claim 43, wherein transferring the sample comprises removing the sample from the cartridge via a port thereon.

45. The method of claim 43, wherein the analytical tool is located in the cartridge, in the instrument, or in an analytical instrument system of the workcell.

46. The method of claim 45, wherein, when the analytical tool is within the analytical instrument system, transferring the sample comprises moving the cartridge to the analytical instrument system via the robotic ami.

47. The method of claim 42 further comprising, prior to transferring the material to the purification module, transferring the material to a digestion module.

48. The method of claim 42 further comprising, prior to transferring the material to the purification module, performing a digestion processing step on the material within the transcription module.

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