A microfluidic system and process for high throughput synthesis and self-contained discrete manufacturing of nanoparticles

Abstract

A system for use in synthesising nanoparticles, the system comprising a microfluidic system reader and a chip cartridge.

Description

[0001] Title

[0002] A microfluidic system and process for high throughput synthesis and self-contained discrete manufacturing of nanoparticles

[0003] Field of the Invention

[0004] The invention relates to a microfluidic system for high throughput screening and optimising of chemical formulations of nanoparticles and subsequent single use discrete production of personalised nanomedicine. More specifically, the invention relates to a high throughput nanoparticle synthesis unit comprising a chip cartridge and a pressure distributor assembly, and a system for high throughput screening and optimisation of various chemical formulations of nanoparticles and discrete personised nanomedicine manufacturing, e.g., lipid / polymer nanoparticles (NPs).

[0005] Background to the Invention

[0006] Nanomedicine in the form of nanoparticles is becoming the most important field in medicine for a variety of applications, such as gene therapy, vaccine development and production, and cell therapy. In 2020, the global nanomedicine market was $212.30 billion (USD), with a compound average growth rate (CAGR) of 9.4%. However, significant challenges remain on efficacy, safety, stability and consistency of formulations, and scale-up production, particularly in the area of personised nanomedicine with discrete, small quantity manufacturing, when translating nanoparticles from formulation into clinical application. This is because nanoparticle formation is highly dependent on the active pharmaceutical ingredients (APIs), carrier compositions, concentrations, and flow and mixing conditions. For example, the development of a lipid nanoparticle (LNP)-based delivery system typically necessitates a substantial amount of screening effort focused on the chemistry of lipids, the lipid-to-cargo ratio, and the overall composition of all lipid components, including ionizable lipids, cholesterol, helper lipids, and PEG-lipids. All these formulation parameters have a critical impact on the physicochemical properties of lipid nanoparticles and the overall therapeutic outcome of the delivery system. Therefore, a significant screening and optimisation effort is required to obtain an optimal formulation.

[0007] Conventional laboratory formulation using pipette mixing for formulation discovery lacks consistency due to human errors and is labour intensive. The traditional multi-step procedures employed in the production of nanoparticles engender large product losses, which is unfavourable when expensive reagents, e.g., ionizable lipids, are used. These bulk formulations involve the production of large-sized, highly variable nanoparticles, thus affecting their functionality. Currently there are two leading companies (Precision NanoSystems Inc (PNI) based in Canada and Unchained labs based in the US) offering microfluidics-based technology to support nanoscale medicine formulation. Dialogue with stakeholders (200+ customer discovery and 100+ one-to-one interviews) validates the urgent market need for an alternative to PNI. This is due to the poor throughput of PNI’s offerings and expensive consumable costs for formulation discovery, and waste. Unchained-labs offers reusable glass chips, but it can cause cross-contamination. There is no effective formulation screening and optimisation system that can meet the customer needs perfectly.

[0008] Bulk methods also lack precise control over the procedure and parameters, thus incurring an increase in cost due to the requirement of additional processing and quality control. Additionally, traditional batch production using large reactors for nanoparticles normally leads to batch-to-batch or intra-batch variability due to heterogeneity from insufficient mixing from fast reactions, e.g., electrostatic interaction for polyplex nanoparticles. As well as the variation of clinical efficacy, there are also safety issues to consider.

[0009] In microfluidics, small volumes of liquid reagents are rapidly mixed in a microchannel in a highly controlled manner to form nanoparticles with tuneable and reproducible structures that can be tailored for drug delivery and that can be directly used for characterisation as well as in vitro studies. To address the challenges set out above, microfluidic technology has been employed in the sector to increase the control over the physical properties of nanoparticles and to utilise low volume formulations.

[0010] The NanoAssemblr® platform from Precision Nanosystems™ is the market’s top product focusing on nanomedicine for gene therapy, cell therapy, and vaccine development. It consists of a specialised system covering formulation development, scale up, and a good manufacturing practice (GMP) system. However, the NanoAssemblr® platform has a very low efficiency in formulation discovery. Their bifurcating microfluidic channels with circular baffle are limited to only synthesis of lipid-based nanoparticles with high flow rates. It is not suitable for small quantity formulation. Also, the syringe-based design also creates large wastage of reagents.

[0011] Another product is the Microfluidizer® high shear fluid processors from Microfluidics International Corporation™. It consists of high shear rate homogenizers by fixed- geometry interaction chambers that induce uniform particle size reduction for nanoparticles thus proving to be suitable for their synthesis and development. The high shear stress used by the homogenizers of the Microfluidizer® can lead to degradation of nucleic acids such as DNA or RNA, thus it is not suitable for the development of nanoparticles for gene therapy. This system also is not suitable for formulation discovery and optimisation, as the single test takes a significant amount of reagent.

[0012] Lastly, Telos® Technology from Dolomite™ consists of standalone pumps, microfluidic chips, and holders for formulation. Their configurable inert chips, complemented with their configurable production systems are used for formulations. However, the system includes very expensive glass chips and their flow focusing channel design has the potential to cause blockage at the junction while synthesising nanoparticles. Also, as cross-contamination from different formulations may occur, cleaning and reuse of the channels are labour intensive and time consuming.

[0013] Although various publications describe the high-throughput combinatorial synthesis of some lipid components, limited information exists on how to incorporate multiple lipid components and prepare lipid nanoparticles (LNPs) in a high-throughput manner. It is obvious that those systems mentioned above can only test one formulation at a time and they also generate a large volume of waste of expensive reagents. They are not suitable for high throughput, small volume formulation, and thus cannot achieve optimal physicochemical properties, determine therapeutic efficacy, and enhance stability across all potential formulation parameter combinations. Additionally, manually preparing a small volume of lipid stock is prone to human error and is also time-consuming. Also, after screening, the development of personised nanomedicines, e.g. cancer vaccines, requires discrete small volume production.

[0014] It is an object of the invention to overcome at least one of the above-mentioned problems.

[0015] Summary of the Invention

[0016] There is an urgent need for a system to overcome the problem of poor throughput for formulation screening and optimisation and discrete self-contained production to meet industrial requirements for fast discovery and manufacturing of nanoparticles for pre-clinical and clinical studies. The proposed high throughput microfluidic-based nanoparticle synthesis and discrete self-contained manufacturing unit and system of the claimed invention are developed to meet these needs. The claimed invention uses an 8 or 12-channel parallel formulation system to facilitate high efficiency and consistency formulation discovery by using a small quantity of reagents without any cross contamination. By incorporating automated sample preparation, the claimed system can carry out 48 formulations / day, which will offer 67% acceleration of overall nanomedicine pre-clinical process, 68% cost saving in chips and materials, and an 80% reduction in manpower. It can also allow a single channel single use self-contained single formulation discrete manufacturing of large amount formulations for scale up. Several advantages of the self-contained manufacturing system are that the system requires no cleaning of the kit between batches, incorporates self-contained cartridges, is traceable, the flow rate of materials can be monitored in real time, it has no cross-contamination, and can be used with in-line quality control and filtration systems. The unit comprising the system and the cartridge also has the advantage of lending itself to be a continuous manufacturing process using discrete cartridges that are integrated with multiple steps, e.g. formulation, dialysis, and in-line characterisation for quality assurance.

[0017] The subject of this application is a high throughput nanoparticle synthesis and discrete self-contained manufacturing unit that comprises a pressure driven microfluidic synthesis platform (with or without automatic liquid handling), a chip cartridge with a micro-mixing microfluidic channel design for insertion inside a moveable platform, and a software embedded system for screening and optimising nanoparticle chemical formulations, e.g., lipid / polymer nanoparticles (NPs).

[0018] The chip cartridge integrates three rows of wells for accommodation of reagent solutions (organic solution and aqueous solution) and synthetic sample solutions, and multiple independent microfluidic channels for nanoparticle synthesis into one chip cartridge. The reagent solutions are driven by air pressure to eliminate additional fluidic connections and dead volumes, contributing to minimize the loss of reagents and potential cross contamination of reagents. The shape of the wells of the chip cartridge can be any shape (for example, rectangle, circle or oval), and the accommodating volume required can be achieved by changing the size of wells (cross-section area and height). The volume of each well can range from microliters to millilitres. Moreover, the distance between two adjacent channels is same with that of adjacent wells of 96 well plate, making it compatible with 96 well plate systems. The chip cartridge is made from polymers (such as polycarbonate (PC), polystyrene (PS), polypropylene (PP), poly(methyl methacrylate) (PMMA), cyclic olefin co / polymer (COC or COP), etc.) through injection moulding, which are bio-compatible, low cost and disposable. The chip cartridge is inserted into a chip cartridge holder of the instrument of the microfluidic system. Reagents can be run in a single or high-throughput (1 to maximum channel) manner into specially designed microfluidic mixing channels which can be controlled by a corresponding solenoid valve connected to each microfluidic channel (the solenoid valve is an electromechanically operated valve that regulates the flow of fluid in the microfluidic channel by opening and closing the channel). Then the nanoparticles are formed via precipitation after effective mixing. For typical lipid nanoparticles, four ingredients (including a cationic / ionizable lipid, 1 ,2-distearoyl-sn-glycero-3-phosphocholine / 1 ,2-dioleoyl-sn-glycero-3-phosphocholine (DSPC / DOPC), polyethylene glycol (PEG)-lipid, and cholesterol) in an organic solution are mixed with a payload (for example, RNAs, small molecules, peptide, proteins) in aqueous solution to form nanoparticles via an electric-static self-assembly process. The mixing process is critical to form high quality nanoparticles. To shorten the time from start to stable flow, it is necessary to use an organic solution for microfluidic channel prefilling before executing the formulation process to exhaust air from the microfluidic channels, thereby reducing the size polydispersity of nanoparticles. During the formulation process, pneumatic pumps apply precise pressures to the wells containing the reagent, pushing the reagents into a microfluidic channel to synthesise the nanoparticles, and yielding synthetic samples in the wells. The precise pressures are controlled by pneumatic pumps and can achieve a precise feedback control of the flow conditions by monitoring the flow rate within the system. A real-time image processing algorithm processes real-time captured images of an inlet well and an outlet well on the chip cartridge and provide flow rate monitoring output. A heating unit with a temperature sensor is integrated into a chip cartridge holder to provide a desired temperature for the system. To ensure consistency within the multiple microfluidic channels, a pressure distributor splits the pressure from the pneumatic pumps equally to each well of each microfluidic channel (just like a branching tree) by keeping the channel lengths the same from the pressure inlets of the pressure distributor to the pressure inlets of the wells.

[0019] Up to at least one to twelve (or more) different formulations ingredients or reagents can be transported to the chip cartridge and then inserted into the moveable platform to synthesis lipid or polymer nanoparticles via a specially designed microfluidic channel with a baffle or an aerofoil structure for efficient mixing. Formulation and flow conditions are easily setup on a touchscreen-based user interface, and the pressure to drive the flow synthesis is automatically calculated by the microfluidic system. After nanoparticle synthesis, the nanoparticles can be characterised physically both in vitro and in vivo for the screening and optimisation for fit to purpose formulations. The claimed system can test multiple formulations of various nanoparticles that are synthesised simultaneously in a highly efficient manner, while controlling the properties of the nanoparticles (for example, size, PDI, encapsulation efficiency) by rapid mixing of components with a much lower waste output.

[0020] To reduce labour time and minimise manual error between batches, a fully automated system can also be used. The automated system of the claimed invention includes a robotic system which handles the transfer of components of the formulations for the drug products being made into the microfluidic chip cartridges for use in the microfluidic system. This microfluidic system, which can be automated, provides a hands-free operation with DoE or machine learning tools embedded for automatic formulation generation.

[0021] In addition, the claimed invention can quickly judge whether the chemical composition is correctly synthesising the desired nanoparticles under typical flow conditions, which can be scaled up.

[0022] In one aspect, there is provided a high throughput nanoparticle synthesis and (discrete) self-contained manufacturing unit as described in the appended claims.

[0023] In one aspect, there is provided a microfluidic system as described in the appended claims.

[0024] In one aspect, there is provided a chip cartridge as described in the appended claims.

[0025] In one aspect, there is provided a method of synthesising nanoparticles using the chip cartridge and the microfluidic system as described in the appended claims.

[0026] In one aspect, there is provided high throughput nanoparticle synthesis and discrete self-contained manufacturing unit comprising (a) a chip cartridge (20) and (b) a microfluidic system (1);

[0027] the chip cartridge (20) comprises a base (21) and a plurality of spaced apart wells (22) there upon, wherein the wells (22) are arranged in a first row (22a), a second row (22b), and a third row (22c); wherein the wells (22) in the first row (22a) act as a first inlet well (24); wherein the wells (22) in the second row (22b) act as a second inlet well (26); wherein the wells (22) of the third row (22c) act as an outlet well (28); and wherein each well (22) comprises at least one microfluidic channel (29), the microfluidic channel (29) comprising a first inlet (29a), a second inlet (29b) and an outlet (29c) which are in fluid communication with the first inlet well (24), the second inlet well (26) and the outlet well (28), respectively; and the microfluidic system (1) comprising a pressure distributor assembly (50), a chip cartridge holder (8), and a monitoring device (12);

[0028] wherein the chip cartridge (20) is sealed by a sealing member (51), the sealing member (51) is selected from a chip cartridge cap (200) or a sealing member holder (53) forming part of the pressure distributor assembly (50); and wherein the sealing member holder (53) comprises a plurality of caps (52) arranged in a first row (52a), a second row (52b) and a third row (52c) configured to engage with the first row (22a), the second row (22b) and the third row (22c), respectively, of the rows (22) of the chip cartridge (20).

[0029] In one aspect, the pressure distributor assembly (50) further comprises a first pressure feed inlet (64), a second pressure feed inlet (66), and a pressure exhaust outlet (68).

[0030] In one aspect, the first row (52a), the second row (52b) and the third row (52c) or the plurality of caps (52) are configured to have a first pressure inlet (54), a second pressure inlet (56) and a third pressure outlet (58), respectively. Preferably, the first pressure inlet (54), the second pressure inlet (56), and the pressure outlet (58) are positioned in the centre of each cap of the plurality of caps (52). In one aspect, the pressure outlet (58) is connected to the pressure exhaust outlet (68). In one aspect, the pressure exhaust outlet (68) is connected to ambient atmosphere via an independent valve.

[0031] In one aspect, the valve is an active valve or a passive valve. Preferably, the active valve is selected from a group comprising a solenoid valve, a pneumatic membrane valve, a piezoelectric valve, and a thermal actuated valve. In one aspect, the passive valve is selected from the group comprising a one-way valve, a check valve, an active valve, a duckbill valve, an umbrella valve, a ball check valve, and a flapper valve. It should be understood that the valves used with the subject invention are not limited to those listed herein, which are presented as examples. In one aspect, the first pressure feed inlet (64) and the second pressure feed inlet (66) apply pressure to the nanoparticle synthesis unit via the first pressure inlet (54) and the second pressure inlet (56), respectively.

[0032] In one aspect, the chip cartridge holder (8) further comprises a base (81) and a heating unit (82) having a heating element and a temperature sensor and a chip cartridge detection sensor (86) adapted to determine the chip cartridge’s (20) position on the base (81).

[0033] In one aspect, the monitoring device (12) comprises at least one camera configured for the real-time volume monitoring of the inlet wells (24,26) and the outlet well (28); and a minimum of one camera configured for monitoring flow in the microfluidic channel (29). In one aspect, the monitoring device (12) comprises at least one camera configured for the real-time volume monitoring of the inlet wells (24,26) and the outlet well (28); and a minimum of one light source configured to illuminate the inlet wells (24,26) and the outlet well (28). In one aspect, both are preferred. Preferably, the monitoring device (12) provides flow rate feedback to a pneumatic pump to control the flow rate in the microfluidic channel (29).

[0034] In one aspect, the synthesis unit according to any one of the preceding claims, wherein the sealing member holder (53) further comprises a sealing member (51). Preferably, the sealing member (51) is a gasket.

[0035] In one aspect, the microfluidic channel (29) is adapted to accommodate aqueous solutions and solvent solutions.

[0036] In one aspect, the first row (22a), the second row (22b) and the third row (22c) have at least one to twenty-four wells. In one aspect, there are at least one to sixteen wells (22). In one aspect, there are at least one to twelve wells (22). In one aspect, there are one to eight wells (22).

[0037] In one aspect, the first row (22a), the second row (22b) and the third row (22c) each have at least one to twenty-four wells. In one aspect, each row has at least one to sixteen wells (22). In one aspect, each row has at least twelve wells (22). In one aspect, each row has at least eight wells (22). In one aspect, there are at least twenty-four wells (22). In theory, the number of wells can be set by the limitation put on that number by the microfluidic channel mould process. In one aspect, the first row (22a), the second row (22b) and the third row (22c) of the chip cartridge (20) have one well with a single microfluidic channel (29). In one aspect, the first row (22a), the second row (22b) and the third row (22c) of the chip cartridge (20) have two wells, each with a single microfluidic channel (29). In one aspect, the first row (22a), the second row (22b) and the third row (22c) of the chip cartridge (20) have three wells with a single microfluidic channel (29). In one aspect, the first row (22a), the second row (22b) and the third row (22c) of the chip cartridge (20) have four wells, each with a single microfluidic channel (29). In one aspect, the first row (22a), the second row (22b) and the third row (22c) of the chip cartridge (20) have five wells, each with a single microfluidic channel (29). In one aspect, the first row (22a), the second row (22b) and the third row (22c) of the chip cartridge (20) have six wells, each with a single microfluidic channel (29). In one aspect, the first row (22a), the second row (22b) and the third row (22c) of the chip cartridge (20) have seven wells, each with a single microfluidic channel (29). In one aspect, the first row (22a), the second row (22b) and the third row (22c) of the chip cartridge (20) have eight wells, each with a single microfluidic channel (29). In one aspect, the first row (22a), the second row (22b) and the third row (22c) of the chip cartridge (20) have nine wells, each with a single microfluidic channel (29). In one aspect, the first row (22a), the second row (22b) and the third row (22c) of the chip cartridge (20) have ten wells, each with a single microfluidic channel (29). In one aspect, the first row (22a), the second row (22b) and the third row (22c) of the chip cartridge (20) have eleven wells, each with a single microfluidic channel (29). In one aspect, the first row (22a), the second row (22b) and the third row (22c) of the chip cartridge (20) have thirteen wells, each with a single microfluidic channel (29). In one aspect, the first row (22a), the second row (22b) and the third row (22c) of the chip cartridge (20) have fourteen wells, each with a single microfluidic channel (29). In one aspect, the first row (22a), the second row (22b) and the third row (22c) of the chip cartridge (20) have fifteen wells, each with a single microfluidic channel (29). In one aspect, the first row (22a), the second row (22b) and the third row (22c) of the chip cartridge (20) have sixteen wells, each with a single microfluidic channel (29). In one aspect, the first row (22a), the second row (22b) and the third row (22c) of the chip cartridge (20) have seventeen wells, each with a single microfluidic channel (29). In one aspect, the first row (22a), the second row (22b) and the third row (22c) of the chip cartridge (20) have eighteen wells, each with a single microfluidic channel (29). In one aspect, the first row (22a), the second row (22b) and the third row (22c) of the chip cartridge (20) have nineteen wells, each with a single microfluidic channel (29). In one aspect, the first row (22a), the second row (22b) and the third row (22c) of the chip cartridge (20) have twenty wells, each with a single microfluidic channel (29). In one aspect, the first row (22a), the second row (22b) and the third row (22c) of the chip cartridge (20) have twenty-one wells, each with a single microfluidic channel (29). In one aspect, the first row (22a), the second row (22b) and the third row (22c) of the chip cartridge (20) have twenty-two wells, each with a single microfluidic channel (29). In one aspect, the first row (22a), the second row (22b) and the third row (22c) of the chip cartridge (20) have twenty-three wells, each with a single microfluidic channel (29). In one aspect, the first row (22a), the second row (22b) and the third row (22c) of the chip cartridge (20) have twenty-four wells, each with a single microfluidic channel (29).

[0038] In one aspect, the first row (22a), the second row (22b) and the third row (22c) have one well with a single microfluidic channel (29). In one aspect, the first row (22a), the second row (22b), and the third row (22c) are covered by the chip cartridge cap (200). In one aspect, the chip cartridge cap (200) is configured to have an active valve or a passive valve. In one aspect, the valve is an active valve or a passive valve. Preferably, the active valve is selected from a group comprising a solenoid valve, a pneumatic membrane valve, a piezoelectric valve, and a thermal actuated valve. In one aspect, the passive valve is selected from the group comprising a one-way valve, a check valve, an active valve, a duckbill valve, an umbrella valve, a ball check valve, and a flapper valve.

[0039] In one aspect, each of the wells (22) are spaced at least 8 mm apart, but no more than about 100 mm apart; or approximately 8 mm to 10 mm apart. In one aspect, each of the wells (22) are spaced approximately 9 mm apart.

[0040] In one aspect, the wells (22) of the chip cartridge (20) are substantially four-sided wells, substantially oval wells, or are substantially circular wells.

[0041] In one aspect, the microfluidic channel (29) leaving the first inlet (29a) is in fluid communication with the first inlet well (24) splits at a junction (29i), and combines with the microfluidic channel (29) connected to the second inlet (29b) at a junction (29j).

[0042] In one aspect, the flow rate is between about 100 ul / min to 100 ml / min. In one aspect, the flow rate is between about 100 ul / min to 90 ml / min. In one aspect, the flow rate is between about 100 ul / min to 75 ml / min. In one aspect, the flow rate is between about 100 ul / min to 50 ml / min. In one aspect, the flow rate is between about 100 ul / min to 40 ml / min. In one aspect, the flow rate is between about 100 ul / min to 30 ml / min. In one aspect, the flow rate is between about 100 ul / min to 20 ml / min. In one aspect, the flow rate is between about 100 ul / min to 10 ml / min. In one aspect, the flow rate is between about 100 ul / min to 5 ml / min. In one aspect, the flow rate is between about 100 ul / min to 1 ml / min.

[0043] In one aspect, the chip cartridge (20) further comprises an automatic identification and data capture technology selected from a radio-frequency identity, a QR code, a bar code, a magnetic strip, and an optical character recognition tag.

[0044] In one aspect, the chip cartridge (20) is made from material selected from the group comprising polycarbonate, polystyrene, polypropylene, poly(methyl methacrylate), cyclic olefin co / polymer, polydimethylsiloxane, thermoset polyester, glass / fused silica, silicon glass hybrids, calcium fluoride, stainless steel (that is GMP compatible), and combinations thereof.

[0045] In one aspect, the chip cartridge cap (200) is manufactured from a material selected from a group comprising Silicone Rubber (VMQ - Vinyl Methyl Silicone Rubber), Fluorosilicone (FVMQ - Fluorovinyl Methyl Silicone Rubber), EPDM (Ethylene Propylene Diene Monomer Rubber), PTFE (Polytetrafluoroethylene), Expanded PTFE (ePTFE -Microporous Polytetrafluoroethylene), FKM (Viton - Fluoroelastomer), FFKM (Kalrez, Chemraz - Perfluoroelastomer), PDMS (Polydimethylsiloxane), PEEK (Polyether Ether Ketone), PFA (Perfluoroalkoxy Alkane), and FEP (Fluorinated Ethylene Propylene).

[0046] In one aspect, the chip cartridge (20) is sterilisable for reuse or is a sterilised single-use unit.

[0047] In one aspect, the chip cartridge (20) has a capacity to hold 50 pl to 1000 ml of the final manufactured product.

[0048] In one aspect, the chip cartridge (20) is a self-contained unit. In one aspect, the chip cartridge (20) is hermetically sealed.

[0049] In one aspect, there is provided a high throughput nanoparticle synthesis unit described herein for use in manufacturing personalised medicines.

[0050] In one aspect, the invention provides a hermetically sealable cartridge for single-use manufacturing of personalised medicine. The cartridge is configured to be filled at a drug and buffer preparation suite with one or more inlet liquids, subsequently hermetically sealed, and transported to a production bioreactor suite for mixing or processing. The sealed configuration prevents contamination by bacteria, moulds, or other pathogens, and prevents leakage of any liquid contents, including through seals, valves, or interfaces, during handling or transportation.

[0051] The cartridge may include one or more inlet valves, which in certain embodiments are passive valves configured to open in response to applied air pressure during liquid injection and automatically close when the internal pressure returns to ambient, thereby maintaining aseptic integrity. In other embodiments, the inlet valves may be active valves actuated by external control mechanisms.

[0052] The cartridge further comprises an outlet valve positioned along an outlet or vent channel, configured to release displaced air during the mixing process and to re-seal automatically after mixing. In this manner, the entire unit remains hermetically sealed post-operation, enabling safe transport and storage of the mixed formulation without risk of contamination or leakage.

[0053] In one aspect, the personalised medicine is selected from the group comprising mRNA therapeutics, siRNA therapeutics, antisense oligonucleotides, CRISPR-based gene editing agents, and plasmid DNA formulations. In one aspect, the medicine is formulated within lipid nanoparticles, polymeric nanoparticles, inorganic nanoparticles, metallic nanoparticles, semiconductor nanoparticles, carbon-based nanoparticles, 2D-material-based nanoparticles, biomimetic nanoparticles, or hybrid nanoformulations configured for targeted delivery.

[0054] The high throughput nanoparticle synthesis unit according to any one of the preceding claims further comprising a console (4) and a services unit (10).

[0055] In one aspect, there is provided a method of synthesising nanoparticles using the high throughput nanoparticle synthesis unit as described herein, the method comprising the steps of: applying an aqueous and an organic solution to the first inlet well (24) and the second inlet well (26), respectively; placing the chip cartridge (20) onto the chip cartridge holder (8) by aligning the base (21) of the chip cartridge (20) with the holder guide (88) on the base (81) of the chip cartridge holder (8); sliding the chip cartridge holder (8) into the bay (6) of the system (1); moving the pressure distributor assembly (50) downwards on a linear motion platform in the microfluidic system (1) so that the sealing member (51) comes into contact with the chip cartridge (20); inputting a synthesis recipe and reagent flow conditions into the microfluidic system (1) via the console (4) on the microfluidic system (1); enabling or disabling the microfluidic channels (29) by opening or closing a corresponding valve when a pre-setting contact force is reached between the sealing member (51) and chip cartridge (20), an air tight seal has been attained; running the synthesis recipe under pressure through the microfluidic channel (29) on the chip cartridge (20) to synthesis the nanoparticles; and collecting the synthesised nanoparticles in the outlet well (28) of the chip cartridge (20) in the microfluidic system (1). In one aspect, the first inlet well (24) and the second inlet well (26) are capped by a cap (52) or the chip cartridge cap (200). In one aspect, the airtight seal is achieved by capping the chip cartridge (20) with the chip cartridge cap (200) or the sealing member holder (53) of the pressure distribution assembly (50). In one aspect, the chip cartridge cap (200) has an active valve or a passive valve. In one aspect, the valve is an active valve or a passive valve. Preferably, the active valve is selected from a group comprising a solenoid valve, a pneumatic membrane valve, a piezoelectric valve, and a thermal actuated valve. In one aspect, the passive valve is selected from the group comprising a one-way valve, a check valve, an active valve, a duckbill valve, an umbrella valve, a ball check valve, and a flapper valve.

[0056] In one aspect, the method further comprises a prefilling step prior to the first step, the prefilling step comprising running a prefilling formulation through the microfluidic channels (29) to exhaust air inside the microfluidic channels (29), and removing any of the formulation remaining in the first inlet well (24), the second inlet well (26), and the outlet well (28). In one aspect, the removal step is performed using a pipette, a pressure driven microfluidic flow, automated robot handling, and / or vacuum suction.

[0057] In one aspect, the aqueous and organic solutions are driven through the microfluidic channel (29) by compressed gas pressure. In one aspect, the gas is selected from air, nitrogen, argon, carbon dioxide, helium, neon, krypton, xenon, and radon.

[0058] In one aspect, the pressure is supplied by a pneumatic pump connected to the first pressure feed inlet (64) and the second pressure feed inlet (66).

[0059] In one aspect, the pressure distributor assembly (50) splits two pressures from the first pressure feed inlet (64) and the second pressure feed inlet (66) equally to rows (22a) and (22b) via the first pressure inlet (54) and the second pressure inlet (56), respectively, by way of a branching tree. In one aspect, the pneumatic pump supplies two pressures from the first pressure feed inlet (64) and the second pressure feed inlet (66) equally to rows (22a) and (22b) via the first pressure inlet (54) and the second pressure inlet (56), respectively, when a single microfluidic channel (29) is used.

[0060] In one aspect, the microfluidic system (1) comprises the housing (2) having a console (4), the bay (6) for receiving the nanoparticle synthesis unit described herein, the movable platform (8) incorporated in the bay (6), a services unit (10), and a control unit comprising: a memory to store one or more computer-executable instructions, and a processor operably coupled to the memory for executing the one or more computerexecutable instructions to display a graphical user interface (GUI) on the console (4), wherein the GUI (30) enables the user to input one or more fluid properties and parameters for the synthesis operation. Preferably, the services unit (10) is configured to control and slide the moveable platform (8) out from within the bay (6) to accommodate the chip cartridge (20).

[0061] In one aspect, the monitoring device (12) relays real-time volume monitoring of the inlet wells (24,26) and the outlet well (28) or the flow in the microfluidic channel (29) to determine the flow rate required in the chip cartridge (20).

[0062] In one aspect, the monitoring device (12) relays real-time flow and output volumes in the microfluidic channel (29) to determine the flow rate required in the chip cartridge (20).

[0063] In one aspect, the console (4) enables the user to set the flow rate, concentration and volume of the first (24) and second (26) input wells of the chip cartridge (20), select a chip cartridge (20) preset parameter and set an output mass ratio.

[0064] In one aspect, the microfluidic system is manually operated or is automated.

[0065] In one aspect, there is provided a single-use, hermetically sealed chip cartridge (20) for use in nanoparticle synthesis or reagent manufacturing, the chip cartridge (20) comprising a base (21) and a plurality of spaced apart wells (22) there upon, wherein the wells (22) are arranged in a first row (22a), a second row (22b), and a third row (22c); wherein the well (22) in the first row (22a) acts as a first inlet well (24); wherein the well (22) in the second row (22b) acts as a second inlet well (26); wherein the well (22) of the third row (22c) acts as an outlet well (28); and wherein each well (22) comprises at least one microfluidic channel (29), the microfluidic channel (29) comprising a first inlet (29a), a second inlet (29b) and an outlet (29c) which are in fluid communication with the first inlet well (24), the second inlet well (26) and the outlet well (28), respectively; wherein the chip cartridge (20) is hermetically sealed by a chip cartridge cap (200).

[0066] In one aspect, the single-use, hermetically sealed chip cartridge (20) described herein further comprises one or more inlet valves (302) selected from an active valve or a passive valve, or a combination thereof.

[0067] In one aspect, the single-use, hermetically sealed chip cartridge (20) described herein, further comprises an outlet valve (304) selected from an active valve or a passive valve.

[0068] In one aspect, the active valve is selected from the group comprising a solenoid valve, a pneumatic membrane valve, a piezoelectric valve, and a thermal actuated valve. In one aspect, the active valve is actuated by an external control mechanism selected from an electrical, a thermal, a magnetic, a piezoelectric or a pneumatic activator.

[0069] In one aspect, the passive valve is selected from the group comprising a one-way valve, a check valve, a passive valve, an active valve, a duckbill valve, an umbrella valve, a ball check valve, and a flapper valve. In one aspect, the passive valve is configured to open in response to an applied gas pressure during liquid injection and to close automatically when the internal pressure returns to ambient pressure.

[0070] In one aspect, the outlet valve (304) is configured to close automatically after a mixing operation, such that the chip cartridge (20) remains hermetically sealed during subsequent transport or storage.

[0071] In one aspect, the active valve or the passive valve, or both, are integrated in the chip cartridge cap (200).

[0072] Some of the advantages of the claimed invention are as follows:

[0073] (1) The claimed invention is designed to allow at least eight, or at least 16, different formulations to be synthesised simultaneously. The consistency and uniformity of the formulations can also be validated.

[0074] (2) The claimed system can run at least eight, or at least 16, formulations simultaneously and separately without any cross-contamination.

[0075] (3) The claimed system can run single or multiple formulations simultaneously and separately without any cross-contamination using a self- contained cartridge. (4) The software embedded system allows the user to enhance control and customisation of NP synthesis by setting reaction parameters, adjusting flow conditions (such as flow rate and flow rate ratio), and can also import / export their flow conditions.

[0076] (5) The chip cartridge designed for use with the claimed microfluidic system removes the need for a physical tubing connection as solutions are placed directly into the chip cartridge, thus minimising waste, eliminating cross contamination, and with a well volume designed for three replications of an in vitro test.

[0077] (6) In addition to minimising reagent waste, the pressure driven microfluidic system also allows zero dead volume in the microfluidic channels of the chip cartridge.

[0078] (7) The microfluidic mixing channels of the claimed invention are optimised for formulation with controlled size and PDI with a total flow rate from 100 uL / min to 50000 uL / min, with a wide flow rate ratio of from 1:1 to 5:1 (aqueous:organic), which is scalable for production.

[0079] (8) The claimed microfluidic channels contain one or more aerofoil and / or rectangular baffle structures to enhance the mixing for synthesis size and uniformity controllable nanoparticles.

[0080] (9) The claimed microfluidic system is designed to be compatible with 96 well plates and 1, 2, 3, 4, 5, 6, 7, 8, 12, 14, 16, or 24 channel pipettes, thus allowing for a seamless integration into the workflow of all testing facilities and for upscaling. (10) The claimed microfluidic system can be incorporated with robotic hands for the automated liquid handling for automated formulation.

[0081] (11) The claimed microfluidic system can be embedded with software for DoE or machine learning to generate formulation combinations automatically for increased high throughput formulation in a batch-by-batch manner.

[0082] (12) The claimed microfluidic system can run single or several channels self- contained cartridge with cap with traceable labels for batch-based production.

[0083] The overall design is innovative and targeted to solve the problem of high throughput effective formulation screening and optimisation in smaller research and development labs to single use discrete production of personalised nanomedicine to large pharmaceutical companies or contract development and manufacturing organisations (CDMOs).

[0084] Definitions In the specification, the term “aerofoil” should be understood to mean a body shaped to produce an aerodynamic reaction (lift) perpendicular to its direction of motion, for a small resistance (drag) force in that plane. The body shape can be, for example, a flat-convex shape (like a lens), a wing, a sail, or the blades of a propeller, a rotor, or a turbine. The upper surface of the aerofoil is generally associated with higher velocity and lower static pressure. The lower surface of the aerofoil has a comparatively higher static pressure than the upper surface. The pressure gradient between these two surfaces contributes to the lift force generated for a given aerofoil.

[0085] In specification, the term “baffle angle” should be understood to mean the angle between a longitudinal direction of the baffle structure situated to the outer profile of a microfluidic channel and the forward flow direction of the fluid.

[0086] In the specification, the term “leading edge” should be understood to be the point at the front of the aerofoil that has maximum curvature (minimum radius).

[0087] In the specification, the term “trailing edge” should be understood to mean the point of maximum curvature at the rear of the aerofoil. The “chord line” is the straight line connecting the leading and trailing edges.

[0088] In the specification, the term “personalised medicine” should be understood as an emerging practice of medicine that delivers a therapy tailored to an individual that ensures a strong or stronger therapeutic response than a generic therapy. As such, the drug substance or substances is formulated into the drug product, such as an LNP, in demand to the individual’s need. This may be determined using an individual's genetic profile, or in response to a diagnostic test, that will guide decisions made in regard to the prevention, diagnosis, and treatment of disease.

[0089] In the specification, the term “self-contained manufacturing unit” should be understood to mean a chip cartridge of the claimed invention that is sealed in an (air-tight) manner to prevent the solutions from interacting with and contamination from the environment or microfluidic system and preventing any cross-contamination between various formulations or batches during the process of manufacturing. Brief Description of the Drawings

[0090] The invention will be more clearly understood from the following description of an embodiment thereof, given by way of example only, with reference to the accompanying drawings, in which: - FIG. 1 illustrates a microfluidic system to run different microfluidic cartridges of the claimed invention.

[0091] FIG. 2 illustrates (A) a perspective view, (B) a plan view, and (C) an underside view of a chip cartridge for use with the microfluidic system of FIG.1 , where the underside view in (C) shows the microfluidic channel design of the chip cartridge; while (D)-(G) illustrate aspects of the cartridge chip (having different volume of wells and channels) that can be used with the microfluidic system of FIG. 1.

[0092] FIG. 3 illustrates a chip cartridge holder with (A) the chip cartridge being inserted therein and (B) with the chip cartridge in situ.

[0093] FIG. 4 illustrates a pressure distributor assembly for use with the microfluidic system of FIG. 1, the chip cartridge of FIG. 2, and the chip cartridge holder of FIG. 3. FIG. 4(A) is a perspective view of the underside of the assembly, while FIG. 4(B) and 4(C) are a front and back view, respectively.

[0094] FIG. 5 illustrates a pressure distribution strategy for exerting equal pressure through all of the microfluidic channels of the chip cartridge.

[0095] FIG. 6 illustrates a perspective view of the pressure distributor assembly of FIG.

[0096] 5 being positioned over the chip cartridge holder of FIG. 3.

[0097] FIG.7 is a cross-section view of FIG. 6 showing the pressure distributor assembly of FIG. 6 in contact with the chip cartridge holder of FIG. 3.

[0098] FIG. 8 illustrates the fully automated (hands-free) version of the microfluidic system to run different single or multiple channel cartridges of FIG. 1.

[0099] FIG. 9 is a flow chart illustrating the workflow of the high throughput nanoparticle synthesis unit comprising the microfluidic system and chip cartridge of the claimed invention.

[0100] Detailed Description of the Drawings

[0101] The invention relates to a high throughput nanoparticle synthesis and discrete self-contained manufacturing unit comprising a microfluidic system that comprises a pressure-driven microfluidic flow synthesis platform and one or more chip cartridges with a micro-mixing channel design for screening chemical formulations of nanoparticles, e.g., lipid or polymer nanoparticles (NPs). The claimed unit adopts a unique high throughput microfluidic based technology fortesting of formulations of lipid / polymer nanoparticles. Referring now to the figures, where FIG. 1 illustrates a general embodiment of a nanoparticle synthesis unit of the present invention for use in formulating, synthesising, and screening nanoparticles. Specifically, FIG. 1 illustrates a front view of the microfluidic system of the present invention and is generally referred to by reference numeral 1. Hereinafter, the microfluidic system will simply be referred to as the “system 1”. The system 1 of the illustrated embodiment comprises a housing 2 having a console 4, a bay 6 for receiving a movable chip cartridge holder 8 with a chip cartridge 20 (examples provided are chip cartridges 20a, 20b, 20c, 20d; see FIG. 2) and a pressure distributor assembly 50 comprising a sealing member 51 (see FIG. 4), a services unit 10 (typically comprising an computer, a linear motion module, and pneumatic pumps), a monitoring device 12, and a door 16 for easy access. The services unit 10 controls and slides the moveable chip cartridge holder 8 out from within the bay 6 and which is adapted to accommodate the chip cartridge 20. The services unit 10 does this by using a series of pneumatic pumps, which are each tailored for a specific use, such as moving the chip cartridge holder 6, push liquid through microfluidic channels in the chip cartridge 20 using air pressure, and the like. The monitoring device 12 can be used for real time monitoring of the formulation synthesis process as well as saving it for future access. The sealing member 51 prevents leakage of liquid when pressure is applied to the liquid in the chip cartridge 20.

[0102] Turning now to FIG.2(A)-(G), the chip cartridge 20 is illustrated in five examples, namely chip cartridge 20a-20d. The chip cartridge 20a, 20b, 20c, 20d comprises a base 21 upon which are positioned three to eight mixing channels (A-H) (for example, see FIG. 2(B) and FIG. 2(E)), which are parallel to each other, and evenly spaced. The chip cartridge 20 can synthesis one or more different formulations at the same time by utilising the different mixing channels (A-H) (see FIG. 2(B)). In FIG. 2(A) and FIG. 2(B), the chip cartridge 20a has a plurality of wells 22 that are four-sided or substantially four-sided, where the top or first row 22a of eight wells acts as a first inlet well 24, with a volume of approximately 170 pl - 1.5 ml and a working volume of approximately 500 pl - 1300 pl. The middle or second row 22b of eight wells act as a second inlet well 26 with a volume of approximately 170 pl - 1.5 ml and a working volume of approximately 50 pl - 1300 pl. The first row 22a of wells usually accommodates an aqueous solution(s) for synthesis of lipid nanoparticles, and the second row 22b of wells usually accommodates an organic solution(s) for synthesis of lipid nanoparticles, while the bottom or a third row 22c of eight wells act as an outlet well 28 with a volume of about 350 pl - 2 ml and a working volume of about 280 l - 1.6 ml, and which acts to collect any nanoparticles that have been synthesised in the chip cartridge 20 in the system 1.

[0103] The chip cartridge 20 can synthesise one or more different formulations at the same time by utilising the different mixing channels (A and B) (see FIG. 2(E)). In FIG. 2(E), the chip cartridge 20c has a plurality of wells 22 that are four-sided or substantially four-sided, where the top or first row 22a of two wells acts as a first inlet well 24, with a volume of approximately 50 ml - 1000 ml and a working volume of approximately 40 ml - 900 ml. The middle or second row 22b of two wells act as a second inlet well 26 with a volume of approximately 50 ml - 1000 ml and a working volume of approximately 40 ml - 900 ml. The first row 22a of wells usually accommodates an aqueous solution(s) for synthesis of lipid nanoparticles, and the second row 22b of wells usually accommodates an organic solution(s) for synthesis of lipid nanoparticles, while the bottom or a third row 22c of two wells act as an outlet well 28 with a volume of 100ml to 2000ml and a working volume of about 80ml to 1800ml, and which acts to collect any nanoparticles that have been synthesised in the chip cartridge 20 in the system 1.

[0104] In FIG. 2(F) and FIG. 2(G), the chip cartridge 20d has a plurality of wells 22 that are foursided or substantially four-sided, where the top or first row 22a of one well acts as a first inlet well 24, with a volume of approximately 50 ml - 1000 ml and a working volume of approximately 40 ml - 900 ml. The middle or second row 22b of one well acts as a second inlet well 26 with a volume of approximately 50 ml - 1000 ml and a working volume of approximately 40 ml - 900 ml. The first row 22a of one well usually accommodates an aqueous solution(s) for synthesis of lipid nanoparticles, and the second row 22b of one well usually accommodates an organic solution(s) for synthesis of lipid nanoparticles, while the bottom or a third row 22c of one well acts as an outlet well 28 with a volume of 100 ml to 2000 ml and a working volume of 80 ml to 1800 ml, and which acts to collect any nanoparticles that have been synthesised in the chip cartridge 20 in the system 1. The cartridge 20d used here can be sterilised and self-contained with a cartridge cap 200 on which a single solenoid valve can be attached, allowing air pressure in while preventing liquid splash. It prevents contamination between batches. The chip cartridge 20 is full traceable using such automatic identification and data capture (AIDC) technologies such as radio-frequency identity (RFIDs), QR codes, bar codes, magnetic strips, optical character recognition (OCR), and the like. It should be noted that these volumes (and number of wells per chip cartridge 20) are open to change in order to facilitate scaling up synthesis of larger volumes of NP solutions. The shape of the wells 22 can be rectangle, circle, or oval according to different volume demands, and the desired accommodation volume can also be easily achieved by changing the size of the wells 22 (cross-section area and height). The volume of the wells 22 can range from microliters to tens to hundreds of millilitres.

[0105] There are a series of parallel independent closed microfluidic channels 29 connected to the inlet wells 24,26 and the outlet well 28 on the other side of the chip cartridge 20 (see FIG. 2(C)). The closed microfluidic channels 29 are usually formed by sealing open microfluidic channels using sealing film. The number of microfluidic channels 29 can be 8, 12, 16, or 24, and the distance between two adjacent microfluidic channels 29 is same with that of adjacent wells of a 96-well plate to make it compatible with 96-well plates using multichannel pipette. The base 21 of the chip cartridge 20 is positioned into the chip cartridge holder 8 of the system 1, and a notch structure 70 along a leading edge of the chip cartridge 20 can be used to ensure that the chip cartridge 20 is positioned correctly in the chip cartridge holder 8 prior to use. After the inlet wells 24,26 are filled with reagents (aqueous and organic solutions), they are placed on the chip cartridge holder 8, which slides into the bay 6. The user can then input the reaction parameters and the flow conditions into the system 1 via the console 4, and the system 1 can start the synthesis of NP solutions that flow through a microfluidic mixing channel 29 on the chip cartridge 20 (see FIG. 2(C)) and can be collected in the outlet well 28.

[0106] In FIG. 2(D), the chip cartridge 20b has square-shaped, or substantially square-shaped, wells 22. The chip cartridge 20b comprises a top or first row 22a of eight wells, which acts as the first inlet well 24 with a volume of 50 pl - 3 ml and a working volume of 40 pl - 2.5 ml, and which is usually used to accommodate aqueous solutions in the case of lipid nanoparticle synthesis. The middle or second row 22b of eight wells act as the second inlet well 26 with a volume of 50 pl - 3 ml and a working volume of 40 pl - 2.5 ml, and which is usually used to accommodate organic solutions in the case of lipid nanoparticle synthesis. A bottom or a third row 22c of eight wells acts as the outlet well 28 with a volume of 1 ml- 6 ml and a working volume of 800 pl - 5.0 ml, where the nanoparticles that have been synthesised in the system 1 are collected. It should be noted that these volumes (and number of wells per chip cartridge) are open to change in order to facilitate scaling up synthesis of larger volumes of NP solutions. The distance between any two adjacent wells in any one of the first row 22a, the second row 22b, or third row 22c is 9 mm, which is the same distance between adjacent wells of a 96-well plate, and the same distance of adjacent channels of an eight-channel pipette. This means that a user can use an eight-channel (or appropriate) pipette to transport the aqueous solutions, organic solutions, and nanoparticle solutions to and from the chip cartridge 20a, 20b.

[0107] In FIG. 2(E), the chip cartridge 20c has rectangular-shaped, or substantially rectangularshaped, wells 22. The chip cartridge 20c comprises the top or first row 22a of two wells acting as a first inlet well 24, with a volume of approximately 50 ml - 1000 ml and a working volume of approximately 40 ml - 900 ml, and which is usually used to accommodate aqueous solutions in the case of lipid nanoparticle synthesis. The middle or second row 22b of two wells act as a second inlet well 26 with a volume of approximately 50 ml - 1000 ml and a working volume of approximately 40 ml - 900 ml, and which is usually used to accommodate organic solutions in the case of lipid nanoparticle synthesis. The bottom or a third row 22c of two wells act as an outlet well 28 with a volume of about 100 ml -2000 ml and a working volume of about 80 ml - 1800 ml, and which acts to collect any nanoparticles that have been synthesised in the chip cartridge 20 in the system 1.

[0108] In FIG. 2(F) and FIG. 2(G), the chip cartridge 20d has rectangular-shaped, or substantially rectangular-shaped, well 22. The chip cartridge 20d comprises the top or first row 22a of one well acting as a first inlet well 24, with a volume of approximately 30 ml - 400 ml and a working volume of approximately 50 ml - 1000 ml, and which is usually used to accommodate aqueous one well wells acts as a second inlet well 26 with a volume of approximately 50 ml - 1000 ml and a working volume of approximately 40 ml - 900 ml, and which is usually used to accommodate organic solutions in the case of lipid nanoparticle synthesis. The bottom or a third row 22d of one well acts as an outlet well 28 with a volume of about 100 ml - 2000 ml and a working volume of about 80ml-1800ml, and which acts to collect any nanoparticles that have been synthesised in the chip cartridge 20 in the system 1.

[0109] In FIG.2(G), the chip cartridge 20d is illustrated with a chip cartridge cap 200 comprising a series of valves 302,304. The chip cartridge cap 200 further comprises one or more inlet valves 302 selected from an active valve or a passive valve, or a combination thereof; and an outlet valve 304 selected from an active valve or a passive valve. The active valve is selected from the group comprising a solenoid valve, a pneumatic membrane valve, a piezoelectric valve, and a thermal actuated valve. The active valve is typically actuated by an external control mechanism selected from an electrical, a thermal, a magnetic, a piezoelectric or a pneumatic activator. The passive valve is selected from the group comprising a one-way valve, a check valve, a passive valve, an active valve, a duckbill valve, an umbrella valve, a ball check valve, and a flapper valve. The passive valve is configured to open in response to an applied gas pressure during liquid injection and to close automatically when the internal pressure returns to ambient pressure. The outlet valve 304 is configured to close automatically after a mixing operation, such that the chip cartridge 20 remains hermetically sealed during subsequent transport or storage. As illustrated here in FIG. 2G, the inlet valves 302 and the outlet valve 304 are integrated in the chip cartridge cap 200. The chip cartridge cap 200 maintains a hermetic seal on the chip cartridge 20 when in use and transport when filled with reagents or final product (a personalised medicine, for example).

[0110] It should be noted that these volumes (and number of wells per chip cartridge 20) are open to change in order to facilitate scaling up synthesis of larger volumes of NP solutions. The chip cartridge 20 can be manifested by any usual process, for example, by injection moulding.

[0111] The chip cartridge 20 further comprises a series of microfluidic channels 29, as shown byway of example in FIG. 2(C), each of the microfluidic channels 29 connecting the first inlet well 24 to the second inlet well 26 and arriving together at the outlet well 28. FIG.

[0112] 2(C) illustrates the microfluidic channel 29 in more detail. The microfluidic channel 29 comprises a first inlet 29a, a second inlet 29b and an outlet 29c. The inlets 29a, 29b and the outlet 29c are in fluid communication with the first inlet well 24, the second inlet well 26 and the outlet well 28, respectively. The microfluidic channel 29 leaving the first inlet 29a that is in fluid communication with the first inlet well 24 splits at a junction 29i, and combines with the microfluidic channel 29 connected to the second inlet 29b at a junction 29j. The microfluidic channel 29 may comprise a series of titled rectangular baffle structures continuous with an inner wall of the microfluidic channel 29 and forming a microfluidic channel within the microfluidic channel 29. The baffle structures stop backflow of solution from the first inlet 29a by increasing the fluid resistance within the microfluidic channel 29. The pressure drop within the microfluidic channel 29 is directly related to the flow rate and the flow resistance of the fluids within the microfluidic channel 29. At the same time, the titled rectangular baffle structures within the microfluidic channel 29 increases the pressure drop of the fluid travelling from the inlets 29a, 29b to the junction 29j. This increase in pressure drop improves the precise control of flowing conditions through pressure. The explanation for this is that for a higher flow resistance, the change in flow rate results in a higher pressure drop, which means a higher sensitivity of pressure to flowrate. The microfluidic channel 29 has a series of aerofoil baffle structures inside the channel to mix solutions by splitting-and-recombining fluids.

[0113] The microfluidic channel 29 is adapted to accommodate aqueous solutions and organic solutions, and further accommodates the aerofoil structure that acts as a further baffle structure. The aerofoil structure comprises an upper camber line, a lower camber line, a mean camber line, a leading edge, and a trailing edge. The aerofoil baffle structure provides very effective mixing of the solutions.

[0114] FIG. 3 illustrates a chip cartridge holder 8 with (A) the chip cartridge 20 being inserted therein and (B) with the chip cartridge 20 in situ. The purpose of the chip cartridge holder 8 is mainly for securing and positioning the chip cartridge 20 in the system 1. The chip cartridge holder 8 comprises a base 81 and a heating unit 82, which can comprise both a heating element and a temperature sensor. The heating unit 82 is embedded into the base 81 of the chip cartridge holder 8 for precise temperature control of solutions in the inlet wells 24,26 and the outlet well 28 of the chip cartridge 20. The chip cartridge holder 8 has a hollowed structure 84 which accommodates the monitoring device 12. The monitoring device 12 comprises at least three cameras, two for the real-time volumes of the inlet wells 24,26 and the outlet well 28 on the chip cartridge 20, and one for monitoring the flow (and output) in the microfluidic channel 29. The monitored flow rates are used as feedback for accurate control of flow rates in the system 1. To make sure of an accurate position of the chip cartridge 20, a chip cartridge detection sensor 86 can be integrated into the chip cartridge holder 8. The chip cartridge 20 slides into the holder 8 by aligning the base 21 of the chip cartridge 20 with a holder guide 88. The chip cartridge detection sensor 86 will detect whether the chip cartridge 20 has accurately reached the correct position in the holder 8. Only when the chip cartridge 20 is well positioned, a user can execute the desired formulation through a graphical user interface (GUI) on the console 4 of the system 1.

[0115] FIG. 4 illustrates a pressure distributor assembly 50 for use with the microfluidic system 1 of FIG. 1, the chip cartridge 20 of FIG. 2, and the chip cartridge holder 8 of FIG. 3. It should be pointed out that the chip cartridge holder 8 can be used with any one of the chip cartridges 20a-d illustrated here. FIG. 4(A) is a perspective view of the underside of the assembly 50, while FIG. 4(B) and 4(C) are a front and back views, respectively. The pressure distributor assembly 50 comprises a sealing member 51, incorporating a plurality of caps 52 that mirror the shape and positioning of the rows 22a, 22b, 22c of the wells in the chip cartridge 20. The caps 52 are arranged in a first row 52a of eight caps that acts as a first pressure inlet 54. The middle or second row 52b of eight caps act as a second pressure inlet 56. The caps 52 arranged on a bottom or a third row 52c act as pressure outlet 58. The rows 52a, 52b and 52c of the pressure distributor assembly 50 correspond to rows 22a, 22b and 22c of the chip cartridge 20, respectively.

[0116] The first pressure inlet 54 and the second pressure inlet 56 have pressure applied to them via a first pressure feed inlet 64 and a second pressure feed inlet 66, respectively. The pressure outlet 58 releases the pressure once the formulations are completed via a plurality of pressure exhaust outlets 68.

[0117] It should be noted that the pressure distributor assembly 50 configuration, including the chip cartridge holder 8 and the sealing member 51 , can be adjusted to match the number of rows 22 in the chip cartridge 20 configuration.

[0118] The pressure distributor assembly 50 can also comprise a sealing member holder 53. When the pressure distributor assembly 50 is installed in the system 1, typically on a linear motion platform which can move up / down (see Arrow A in FIG. 6), the pressure distributor assembly 50 moves downwards, and the sealing member 51 will come into contact with the chip cartridge 20, achieving a full seal between the chip cartridge 20 and the pressure distributor assembly 50 until a pre-setting contact force is reached between the sealing member 51 and chip cartridge 20 (see FIG.6, FIG.7). Once the chip cartridge 20 is well sealed, pressures go into the pressure distributor assembly 50 (see FIG. 5).

[0119] The pressure distributor assembly 50 splits two pressures from the first pressure feed inlet 64 and the second pressure feed inlet 66 equally to rows 22a and 22b via the first pressure inlet 54 and the second pressure inlet 56, respectively, in the way of branching tree (see FIG. 5). The first pressure feed inlet 64 and the second pressure feed inlet 66 are connected to a pneumatic pump. The branching of the supply of pressure applied from the first and second pressure feed inlets 64,66 along the length of the microfluidic channel 29 to the first pressure inlet 54 and the second pressure inlet 56 in the inlet wells 24,26, respectively, remains consistent throughout and along the length of each microfluidic channel 29. The sealing member 51 in the sealing member holder 53 is integrated into the pressure distributor assembly 50. The sealing member 51 is typically made of elastic material, such as silicon rubber, and can incorporate the plurality of caps 52 which align with the corresponding wells 22 of the chip cartridge 20 for a full seal.

[0120] As shown in FIG. 4(A), the first pressure inlet 54, the second pressure inlet 56, and the pressure outlet 58 are positioned in the centre of each cap 52. FIG. 7 shows that when the pressure distributor assembly 50 of FIG. 6 is in contact with the chip cartridge holder 20 of FIG. 3, the first pressure inlet 54, the second pressure inlet 56, and the pressure outlet 58 line up with, and seal, the first inlet well 24, the second inlet well 26, and the outlet well 28, respectively. Each of the pressure outlets 58 is connected to the ambient atmosphere through an independent solenoid valve, so that each microfluidic channel 29 can be opened and closed by opening and closing a corresponding solenoid valve. For the enabled microfluidic channel 29 with its solenoid valve open, the reagents in the first inlet well 24 and the second inlet well 26 will be pushed into the microfluidic channel 29 for mixing, and yield a sample solution with synthesised nanoparticles in the outlet well 28 (see FIG. 7). For the microfluidic channel 29 that has its solenoid valve closed, the microfluidic channel 29 is blocked because the corresponding exhaust outlet 68 is closed.

[0121] An optional first step is to prefill the microfluidic channels 29 with an organic solvent by running a prefilling formulation. The prefilling formulation exhausts the air inside the microfluidic channels 29, which can reduce the time to achieve stable flow during following the formulation process. The running time of the prefilling formulation process is shorter than that of the normal formulation process to ensure that the liquid in the first inlet well 24 and the second inlet well 26 does not run out, and so air will not enter the microfluidic channels 29. After finishing the prefilling formulation process, the residual liquid in the first inlet well 24 and the second inlet well 26, and liquid in outlet well 28 should be removed using a pipette, and the prefilled chip cartridge 20 is ready for the formulation process.

[0122] For the sample preparation of a chosen formulation, an organic solution with a lipid (or a working solution with a polymer) and an aqueous solution with a payload, e.g. DNA / RNA, small molecules (or a working solution with payload), are prepared, and are transported to the first inlet wells 24 and the second inlet wells 26 of the chip cartridge 20, at the volume ratio equal to a desired flow rate ratio. Then, the user can insert the chip cartridge 20 into the chip cartridge holder 8 into the bay 6 of the system 1. The chip cartridge detection sensor 86 detects whether the chip cartridge 20 is in the desired position and gives positive feedback to the user. Only when the chip cartridge 20 is in the desired position does the formulation process proceed and the system 1 can then synthesis lipid / polymer nanoparticles by the flow of the reagents through the microfluidic channels 29 of the chip cartridge 20, driven by compressed air pressure, which is calculated according to the desired flow conditions. The system 1 further includes a control unit (not shown) for controlling and managing the operation of the system 1 for synthesising the nanoparticles. The control unit may represent a computational platform that includes components that may be in a server or another computer system, and execute, by way of a processor (e.g., a single or multiple processors) or other hardware described herein. The methods, functions and other processes of the present invention may be embodied as machine-readable instructions stored on a non-transitory computer-readable medium for providing the GUI on the console 4 (see, FIG. 1) to enable a user / operator to manage the overall synthesis operation, and set a flow rate.

[0123] To execute the desired formulation, the user can optionally enable the prefilled microfluidic channels 29 on the GUI of the interface 4, and then load either a stored formulation recipe or create a new formulation recipe to be executed, and then start the formulation process. The formulation parameters include total flow rate, total volume, flow rate ratio and temperature, as well as the reagents information, such as reagent type, concentrations, etc. During execution of the formulation process, the pressure distributor assembly 50 moves downwards, and the sealing member 51 will come into contact with the chip cartridge 20, achieving a full seal of the wells 24,26,28 of the chip cartridge 20 until a pre-setting contact force is reached between the sealing member 51 and the chip cartridge 20. Once the pressure distributor assembly 50 and the chip cartridge 20 form a tight seal, the pneumatic pump starts to work. To achieve an accurate flow condition during the formulation process, a closed-loop control strategy is adopted here, where real flow rates are monitored by monitoring device 12 to provide feedback on the progression of the formulation and aid in controlling the pressure within the system 1. Flow rate monitoring is achieved by processing real-time images of the first and second inlet wells 24,26. Specifically, real-time images of the first inlet well 24 and the second inlet well 26 are captured using high-resolution cameras, and the volumes of the first inlet well 24 and the second inlet well 26 are identified based on the level of the liquid therein. Then, the flow rates of the fist inlet well 24 and the second inlet well 26 are calculated based on the changes in volume overtime. After the all the reagent solutions in the first inlet well 24 and the second inlet well 26 run out, the pneumatic pump stops to work, and then the sealing member 51 detaches from the chip cartridge 20 or cartridge cap 50 as the pressure distributor assembly 50 moves upwards. When the pressure distributor assembly 50 returns to its initial position, the formulation process ends. Once the synthesis process is completed, lipid / polymer nanoparticles are collected from the outlet well 28 of the chip cartridge 20, which can be then characterised for size, PDI, and encapsulation efficiency. The synthesised nanoparticles can then be used for in vitro or in vivo experiments. Dynamic light scattering (DLS) can be used to measure the size and PDI of the nanoparticles, and cell experiments also can be involved.

[0124] The running information of the formulation process, such as monitoring flow rate, temperature, pressure values, etc., is automatically saved as a running log file for a good traceability of the formulation process for a given formulation. At different stages of the formulation process (like start, operation, and completion), the indication information for the user is displayed on the GUI of the interface 4, and the operation status of the system 1 is also indicated by the colour of an operation indicator light 7 on the housing 2 of the system 1.

[0125] The sample preparation and liquid handling processes involved in the workflow can be done automatically by equipping the system 1 with an automated liquid handling system.

[0126] FIG. 8 illustrates a general embodiment of the fully automated version of the system 1. This automated system 1 further comprises a robotic arm 112 that performs all the operations, making it a hands-free system. The robotic arm 112 has the option to connect pipette tips of various volumes with an automated air displacement pipettor, as well as hold and carry the chip cartridge 20. Pipette tips 103 (different sizes and volumes) will be taken by the robotic arm 112 to collect the required solvents from solvent containers which are placed in solvent holders 105,106. The solvent holder 105 is a temperature-controlled holder where the solvents can be heated or cooled to a specific temperature to maintain the stability of the solvent. Solvents that do not require temperature regulation are kept in the solvent holder 106. Collected solvents are transferred to a 96-well plate 101 which is placed in a shaker module 104 that mixes the solvents properly by shaking at a specific revolutions per minute (RPM). The mixed solution and other solvents are transferred to the first and second inlet wells 24,26, respectively, of the chip cartridge 20, which is kept on a chip cartridge stand 110. The filled chip cartridge 20 is placed on the chip cartridge holder 8 of the system 1 by the robotic arm 112. The liquid in the first and second inlet wells 24,26, respectively, are pushed through the microfluidic channels 29 by applying pressure. The pressure distributor assembly 50 prevents leakage from the first and second inlet wells 24,26 during this process. The synthesised solution is collected in the outlet well 28 which is transferred to another 96-well plate 108 placed on a temperature regulated plate holder 107 by the robotic arm 112. The robotic arm 112 will repeat this process for all the chip cartridges 20 which are placed on the chip cartridge stand 110, thereby facilitating the preparation of many batches quickly. Each chip cartridge 20 is washed using specific solvents before synthesising the formulations. All the waste liquid is collected in a waste container 109. The pipette tips are discarded in a waste box 102 after each use. The liquid flow through the microfluidic channels 29 and their mixing can be viewed or saved using the monitoring device 12.

[0127] FIG. 9 is a flow chart illustrating the workflow 900 of the high throughput nanoparticle synthesis unit comprising the microfluidic system 1 and chip cartridge 20 of the claimed invention.

[0128] At step 902, the wells for example, the first and second inlet wells 24,26 of the chip cartridge 20 of the system 1, are filled with reagent solutions (aqueous solutions).

[0129] At step 904, it is checked if the chip cartridge 20 is present after the chip cartridge 20 has been loaded onto the chip cartridge holder 8. At step 906, it is determined if the chip cartridge 20 is ready. In an embodiment of the present invention, a chip cartridge detection sensor 86 may be provided to detect whether the chip cartridge 20 has accurately reached the correct position in the chip cartridge holder 8. Only when the chip cartridge 20 is well positioned is the chip cartridge 20 ready, and a user can execute the desired formulation through a graphical user interface (GUI) 901 of the system 1. In an embodiment of the present invention, the chip cartridge status may be automatically or manually communicated to the GUI 901 when the chip cartridge 20 is not ready.

[0130] At step 908, the user enables the prefilled microfluidic channels 29 through the GUI 901. The status of the microfluidic channels 29 as ‘enabled channels’ may be automatically or manually communicated to the GUI 901 after the microfluidic channels 29 are enabled.

[0131] At step 910, it is checked if the microfluidic channels 29 are prefilled. If the microfluidic channels 29 are not prefilled, then the user may use the GUI 901 at step 912 to load a prefilling formulation. At step 913, prefilling of the microfluidic channels 29 with an organic solvent is executed by running a prefilling formulation. Upon execution of the prefilling, the workflow goes back to step 902.

[0132] Alternatively, if the microfluidic channels 29 are prefilled, then at step 914, it is checked if a formulation already exist. If a formulation already exists, then at step 916, the user may use the GUI 901 to load an existing formulation, or alternatively, use the GUI 901 at step 918 to create a new formulation recipe with formulation settings 920. The formulation settings 920 include, but are not limited to, total flow rate, total volume, flow rate ratio and temperature, as well as the reagents information, such as reagent type, concentrations, etc. The formulation parameters 920 may be stored in a memory storage 921 , which is communicatively coupled to the GUI 901.

[0133] Alternatively, if the microfluidic channels 29 are not prefilled, then at step 914, it is checked if a formulation already exist. If a formulation already exists, then at step 916, the user may use the GUI 901 to load an existing formulation, or alternatively, use the GUI 901 at step 918 to create a new formulation recipe with formulation settings 920. The formulation settings 920 include, but are not limited to, total flow rate, total volume, flow rate ratio and temperature, as well as the reagents information, such as reagent type, concentrations, etc. The formulation parameters 920 may be stored in a memory storage 921 , which is communicatively coupled to the GUI 901.

[0134] Upon either loading of an existing formulation or creation of new formulation, the formulation process is executed / started at step 922 to generate synthesized samples at step 923. Also, the formulation outcome may be automatically or manually communicated to the GUI 901.

[0135] Further, at step 924, the conditions of the formulation process are continuously monitored to provide feedback on the progression of the formulation and aid in controlling the pressure within the system. Examples of the conditions that are monitored include, but are not limited to, flow rates, remaining volumes, pressure values, temperature, and running times. At step 926, the running information of the formulation process, such as monitoring flow rate, temperature, pressure values, etc., is automatically saved as a running log file for a good traceability of the formulation process for a given formulation. Conclusion

[0136] One of the advantages of the claimed system is that it simplifies the operation processes and up to sixteen different formulations can be synthesised simultaneously. It can be also used as a standalone system to run single formulation using discrete batches with self-contained cartridges for personalised nanomedicine manufacturing.

[0137] The system 1 uses a pneumatic pump and is also easy to set-up for experiments with significant flexibility to change the chip cartridges 20 to adapt different volume and throughput. The system 1 automatically calculates the air pressure according to the set flow parameters, allowing more flexible control of flow conditions for formulation optimisation. The allowed total flow rate ranges from WOuL / min to 50,000ul_ / min with a wide flow rate ratio from 1:1 to 5:1 (aqueous:organic).

[0138] The fully automated system also offers many advantages like reduced labour time, minimise manual error, etc and can prepare different formulations with great precision and high throughput.

[0139] The chip cartridge 20 of the claimed invention and used in the claimed system 1 has the following advantageous features: 1) it is compatible with different organic solvents; 2) it is compatible for use with single channel and multi-channel pipettes, such as an eight, twelve, fourteen, or sixteen channel pipette; 3) it has suitable capacity for optimising flow rate; 4) it minimises reagent loss; 5) it eliminates cross-contamination; 6) it allows a wide latitude in flow rates, from low flow rates (WOuL / min) to high flow rates (50,000ul_ / min), by using different well sizes and applied pressure (flow rate can be changed by changing the air pressure, allowing the user to optimise the flow conditions to get better results); and 7) it is compatible for scaling up the nanoparticle manufacturing process with self-contained single use discrete batch production. The chip cartridge 20 is designed to have at least two inlet wells 24,26 and at least one outlet well 28. Since there are no fluidic connections, the waste is minimised or is non-existent. The rows of wells 22a, 22b of the chip cartridge 20 can optionally be prefilled with solutions before being placed in the system 1. The solution flow is achieved by application of air pressure for which a pneumatic pump is used. This setup avoids the need of any tubing or connectors thereby minimising the wastage due to dead volume (unused volume left in the tubes and connectors). The distance between the wells 22 is 9 mm, which is the same distance between the wells of a 96-well plate and 8-16 channel pipettes. This means that a standard eight-sixteen channel pipette can be used when taking up and dispensing solutions, making the process much more convenient. 2D flow focusing can also be used with the chip cartridge 20, which allows the organic solvents and aqueous solutions mix better. The total flow rate (TFF) can be selected from 100 pl / min to 50,000 pl / min, and the flow rate ratio (FRR) between aqueous and organic solvents can be selected from 1:1 to 5: 1 , in order to optimise flow conditions.

[0140] For the claimed chip cartridge 20 liquid is prevented from splashing the sides of the well and / or from escaping from the well when being applied, by using a working volume that is less than the volume of the well. The working volume of the wells is designed to generate sufficient volume of nanoparticles for both physical characterisation and in vitro test. When the user wishes to scale up to larger volumes of nanoparticles, microfluidic chip cartridges can be manufactured with larger inlet and outlet volumes, up to hundreds of millilitres and liters. The cartridge can be labelled with AIDC methods for trackability and is sterilized. They can be a single channel for large volume or several channels. Dynamic light scattering (DLS) can be used to measure the size and PDI of the nanoparticles, and cell experiments also can be involved. An in-line quality monitoring system can be adapted with inline buffer exchange or filtration for small to large volume discrete batch-based manufacturing.

[0141] Apart from using the system of the claimed invention for formulation discovery and preliminary chemistry selection, in vitro studies can be easily performed due to its compatibility with an 8-16 channel pipette. By discovering a new nanomedicine formulation and optimising it for good efficacy and stability in a way that is dramatically more efficient than existing companies, the system of the claimed invention will support the high-speed development of better pipelines of nanoparticles that are more likely to meet the stringent clinical and manufacturing requirements for new therapeutics, e.g. personalised nanomedicine. In doing so, this can lead to the access of new healthcare options for improved quality of life, particularly like gene therapy (e.g., Alzheimer's disease), vaccine development (e.g., RNA vaccine manufacture against, such as, coronavirus disease (COVID-19) caused by the SARS-CoV-2 virus), and cell therapies (e.g., T-cell cancer therapies).

[0142] A quick response and mitigation of risks to public health, for example, through the rapid development of healthcare options, such as mRNA vaccines for COVID-19, can be greatly improved using the system of the claimed invention. Miniaturisation, and more importantly ‘getting it right first time’, through microscale reaction and precision control of flow conditions and reagent formulation / concentration using the system of the claimed invention can reduce (i) the risk of failure, (ii) the number of optimisation cycles, (iii) the number of current resources required, and (iv) the carbon footprint and overall waste that is generated by current approaches.

[0143] Further, the microfluidic-based flow synthesis platform of the system 1 of the claimed invention can not only be to use to synthesise and develop lipid and polymer nanoparticles, but also extend its application for the continuous, unseeded synthesis of gold nanoparticles, functionalised silica nanoparticles, solid lipid nanoparticles, and liposomes and polymersomes for various therapeutic and diagnostic uses. It can also be used for generation of droplets and prepare samples for life science research.

[0144] In the specification the terms "comprise, comprises, comprised and comprising" or any variation thereof and the terms “include, includes, included and including" or any variation thereof are considered to be totally interchangeable and they should all be afforded the widest possible interpretation and vice versa.

[0145] The invention is not limited to the embodiments hereinbefore described but may be varied in both construction and detail.

[0146] All publications, patents, patent applications and other references mentioned herein are hereby incorporated by reference in their entireties for all purposes as if each individual publication, patent or patent application were specifically and individually indicated to be incorporated by reference and the content thereof recited in full.

Claims

Claims1. A high throughput nanoparticle synthesis and self-contained manufacturing unit comprising (a) a chip cartridge (20) and (b) a microfluidic system (1);the chip cartridge (20) comprises a base (21) and a plurality of spaced apart wells (22) there upon, wherein the wells (22) are arranged in a first row (22a), a second row (22b), and a third row (22c); wherein the well (22) in the first row (22a) acts as a first inlet well (24); wherein the well (22) in the second row (22b) acts as a second inlet well (26); wherein the well (22) of the third row (22c) acts as an outlet well (28); and wherein each well (22) comprises at least one microfluidic channel (29), the microfluidic channel (29) comprising a first inlet (29a), a second inlet (29b) and an outlet (29c) which are in fluid communication with the first inlet well (24), the second inlet well (26) and the outlet well (28), respectively;the microfluidic system (1) comprising a pressure distributor assembly (50), a chip cartridge holder (8) and a monitoring device (12);wherein the chip cartridge (20) is sealed by a sealing member (51), the sealing member (51) is selected from a chip cartridge cap (200) or a sealing member holder (53) forming part of the pressure distributor assembly (50); andwherein the sealing member holder (53) comprises a plurality of caps (52) arranged in a first row (52a), a second row (52b) and a third row (52c) configured to engage with the first row (22a), the second row (22b) and the third row (22c), respectively, of the rows (22) of the chip cartridge (20).

2. The high throughput nanoparticle synthesis unit according to Claim 1, wherein pressure distributor assembly (50) further comprises a first pressure feed inlet (64), a second pressure feed inlet (66), and a pressure exhaust outlet (68).

3. The high throughput nanoparticle synthesis unit according to Claim 1 or Claim 2, wherein the first row (52a), the second row (52b) and the third row (52c) or the plurality of caps (52) are configured to have a first pressure inlet (54), a second pressure inlet (56) and a third pressure outlet (58), respectively.

4. The high throughput nanoparticle synthesis unit according to Claim 3, wherein the first pressure inlet (54), the second pressure inlet (56), and the pressure outlet (58) are positioned in the centre of each cap of the plurality of caps (52).

5. The high throughput nanoparticle synthesis unit according to Claim 3 and Claim 4, wherein the pressure outlet (58) is connected to the pressure exhaust outlet (68).

6. The high throughput nanoparticle synthesis unit according to any one of Claims 2 to 5, wherein the pressure exhaust outlet (68) is connected to ambient atmosphere via an independent valve.

7. The high throughput nanoparticle synthesis unit according to Claim 6, wherein the valve is selected from an active valve or a passive valve.

8. The high throughput nanoparticle synthesis unit according to Claim 7, wherein the active valve is selected from a group comprising a solenoid valve, a pneumatic membrane valve, a piezoelectric valve, and a thermal actuated valve.

9. The high throughput nanoparticle synthesis unit according to Claim 7, wherein the passive valve is selected from a group comprising a one-way valve, a check valve, a duckbill valve, an umbrella valve, a ball checkvalve, and a flapper valve.

10. The high throughput nanoparticle synthesis unit according to any one of Claims 2 to 9, wherein the first pressure feed inlet (64) and the second pressure feed inlet (66) apply pressure to the nanoparticle synthesis unit via the first pressure inlet (54) and the second pressure inlet (56), respectively.

11. The high throughput nanoparticle synthesis unit according to any one of the preceding claims, wherein the chip cartridge holder (8) further comprises a base (81) and a heating unit (82) having a heating element and a temperature sensor and a chip cartridge detection sensor (86) adapted to determine the chip cartridge’s (20) position on the base (81).

12. The high throughput nanoparticle synthesis unit according to any one of the preceding claims, wherein the monitoring device (12) comprises at least one camera configured for the real-time volume monitoring of the inlet wells (24,26) and the outlet well (28); and (a) a minimum of one camera configured for monitoring flow in the microfluidic channel (29) or (b) a minimum of one light source configured for illuminate the inlet wells (24,26) and the outlet well (28), or a combination thereof.

13. The high throughput nanoparticle synthesis unit according to Claim 9, wherein the monitoring device (12) provides flow rate feedback to a pneumatic pump to control the flow rate in the microfluidic channel (29).

14. The high throughput nanoparticle synthesis unit according to any one of the preceding claims, wherein the sealing member holder (53) further comprises a gasket.

15. The high throughput nanoparticle synthesis unit according to Claim 14, wherein the gasket surrounds the plurality of caps (52) or encapsulates the plurality of caps (52).

16. The high throughput nanoparticle synthesis unit according to any one of the preceding claims, wherein the chip cartridge cap (200) is manufactured from a material selected from a group comprising silicone, fluorosilicone, ethylene propylene diene monomer, polytetrafluoroethylene, expanded polytetrafluoroethylene, fluoroelastomer, perfluoroelastomer, polydimethylsiloxane, polyether ether ketone, perfluoroalkoxy alkane, and fluorinated ethylene propylene.

17. The high throughput nanoparticle synthesis unit according to any one of the preceding claims, wherein the microfluidic channel (29) is adapted to accommodate aqueous solutions and solvent solutions.

18. The high throughput nanoparticle synthesis unit according to any one of the preceding claims, wherein the first row (22a), the second row (22b) and the third row (22c) have between one and twenty-four wells (22).

19. The high throughput nanoparticle synthesis unit according to any one of the preceding claims, wherein the first row (22a), the second row (22b) and the third row (22c) each have at least twenty-four wells (22).

20. The high throughput nanoparticle synthesis unit according to any one of the Claims 1 to 18, wherein the first row (22a), the second row (22b) and the third row (22c) each have at least sixteen wells (22).

21. The high throughput nanoparticle synthesis unit according to any one of the preceding claims, wherein the first row (22a), the second row (22b) and the third row22. The high throughput nanoparticle synthesis unit according to any one of the preceding claims, wherein the first row (22a), the second row (22b), and the third row (22c) of the chip cartridge (20) are covered by the chip cartridge cap (200).

23. The high throughput nanoparticle synthesis unit according to Claim 22, wherein the chip cartridge cap (200) is configured to have an active valve or a passive valve.

24. The high throughput nanoparticle synthesis unit according to Claim 23, wherein the active valve is selected from the group comprising a solenoid valve, a pneumatic membrane valve, a piezoelectric valve, and a thermal actuated valve.

25. The high throughput nanoparticle synthesis unit according to Claim 23, wherein the passive valve is selected from the group comprising a one-way valve, a check valve, a duckbill valve, an umbrella valve, a ball check valve, and a flapper valve.

26. The high throughput nanoparticle synthesis unit according to any one of the preceding claims, wherein each of the wells (22) of the chip cartridge (20) are spaced at least 8 mm apart.

27. The high throughput nanoparticle synthesis unit according to Claim 26, wherein each of the wells (22) of the chip cartridge (20) are spaced approximately 9 mm apart.

28. The high throughput nanoparticle synthesis unit according to any one of the preceding claims, wherein the wells (22) of the chip cartridge (20) are substantially foursided wells, substantially oval wells, or are substantially circular wells.

29. The high throughput nanoparticle synthesis unit according to any one of the preceding claims, wherein the microfluidic channel (29) leaving the first inlet (29a) is in fluid communication with the first inlet well (24) splits at a junction (29i), and combines with the microfluidic channel (29) connected to the second inlet (29b) at a junction (29j).

30. The high throughput nanoparticle synthesis unit according to any one of the preceding claims, wherein the flow rate is between about 100 ul / min to 100 ml / min.

31. The high throughput nanoparticle synthesis unit according to any one of the preceding claims, wherein the chip cartridge (20) further comprises an automatic identification and data capture technology selected from a radio-frequency identity, a QR code, a bar code, a magnetic strip, and an optical character recognition tag.

32. The high throughput nanoparticle synthesis unit according to any one of the preceding claims, wherein the chip cartridge (20) is made from material selected from the group comprising polycarbonate, polystyrene, polypropylene, poly(methyl methacrylate), cyclic olefin co / polymer, polydimethylsiloxane, thermoset polyester, glass / fused silica, silicon glass hybrids, calcium fluoride, and stainless steel.

33. The high throughput nanoparticle synthesis unit according to any one of the preceding claims, wherein the chip cartridge (20) is sterilisable for reuse or is a sterilised single-use unit.

34. The high throughput nanoparticle synthesis unit according to any one of the preceding claims, wherein the chip cartridge (20) has a capacity to hold 50 pl to 1000ml of the final manufactured product.

35. The high throughput nanoparticle synthesis unit according to any one of the preceding claims, wherein the chip cartridge (20) is a self-contained unit.

36. The high throughput nanoparticle synthesis unit according to any one of the preceding claims further comprising a console (4) and a services unit (10).

37. The high throughput nanoparticle synthesis unit according to any one of the preceding claims for use in manufacturing personalised medicines.

38. The high throughput nanoparticle synthesis unit according to Claim 37, wherein the personalised medicine is selected from the group comprising mRNA therapeutics, siRNA therapeutics, antisense oligonucleotides, CRISPR-based gene editing agents, and plasmid DNA formulations.

39. The high throughput nanoparticle synthesis unit according to Claim 38, wherein the personalised medicine is formulated within lipid nanoparticles, polymeric nanoparticles, inorganic nanoparticles, metallic nanoparticles, semiconductor nanoparticles, carbon-based nanoparticles, 2D-material-based nanoparticles, biomimetic nanoparticles, or hybrid nanoformulations configured for targeted delivery.

40. A method of synthesising nanoparticles using the high throughput nanoparticle synthesis unit of any one of the preceding claims, the method comprising the steps of:(a) applying an aqueous and an organic solution to the first inlet well (24) and the second inlet well (26), respectively, of the chip cartridge (20);(b) placing the chip cartridge (20) onto the chip cartridge holder (8) by aligning the base (21) of the chip cartridge (20) with the holder guide (88) on the base (81) of the chip cartridge holder (8);(c) sliding the chip cartridge holder (8) into the bay (6) of the microfluidic system (1):(d) moving the pressure distributor assembly (50) downwards on a linear motion platform in the microfluidic system (1) so that the sealing member (51) comes into contact with the chip cartridge (20);(e) inputting a synthesis recipe and reagent flow conditions into the system (1) via the console (4) on the microfluidic system (1);(f) enabling or disabling the microfluidic channels (29) by opening or closing a corresponding valve when a pre-setting contact force is reached between the sealing member (51) and chip cartridge (20), an airtight seal has been attained;(g) running the synthesis recipe under pressure through the microfluidic channel (29) on the chip cartridge (20) to synthesis the nanoparticles; and (h) collecting the synthesised nanoparticles in the outlet well (28) of the chip cartridge (20) in the microfluidic system (1).

41. The method of Claim 40, further comprising an optional prefilling step prior to step (a) comprising running a prefilling formulation through the microfluidic channels (29) to exhaust air inside the microfluidic channels (29), and removing any of the formulation remaining in the first inlet well (24), the second inlet well (26), and the outlet well (28) of the chip cartridge (20).

42. The method according to Claim 40 or Claim 41 , wherein the aqueous and organic solutions are driven through the microfluidic channel (29) by compressed gas pressure.

43. The method according to Claim 42, wherein the gas is selected from air, nitrogen, argon, carbon dioxide, helium, neon, krypton, xenon, and radon.

44. The method according to Claim 42 and 43, wherein the compressed gas pressure is supplied by a pneumatic pump connected to the first pressure feed inlet (64) and the second pressure feed inlet (66).

45. The method according to Claim 44, wherein the pneumatic pump supplies two pressures from the first pressure feed inlet (64) and the second pressure feed inlet (66)equally to rows (22a) and (22b) of the chip cartridge (20) via the first pressure inlet (54) and the second pressure inlet (56), respectively, when a single microfluidic channel (29) is used.

46. The method according to any one of Claims 40 to 45, wherein the pressure distributor assembly (50) splits two pressures from the first pressure feed inlet (64) and the second pressure feed inlet (66) equally to rows (22a) and (22b) of the chip cartridge (20) via the first pressure inlet (54) and the second pressure inlet (56), respectively, by way of a branching tree.

47. The method according to any one of Claims 40 to 46, wherein the microfluidic system (1) of Claim 1 comprises the housing (2) having a console (4), the bay (6) for receiving the nanoparticle synthesis unit of Claim 1, the movable platform (8) incorporated in the bay (6), a services unit (10), and a control unit comprising: a memory to store one or more computer-executable instructions, and a processor operably coupled to the memory for executing the one or more computer-executable instructions to display a graphical user interface (GUI) on the console (4), wherein the GUI (30) enables the user to input one or more fluid properties and parameters for the synthesis operation.

48. The method according to Claim 47, wherein the console (4) enables the user to set the flow rate, concentration, and volume of the first input well (24) and the second input well (26) of the chip cartridge (20), select a chip cartridge preset parameter, and set an output mass ratio.

49. The method according to any one of Claims 40 to 48, wherein the monitoring device (12) relays real-time volume monitoring of the first and second inlet wells (24,26) and the outlet well (28) or the flow in the microfluidic channel (29) to determine the flow rate required in the chip cartridge (20).

50. The method according to any one of Claims 40 to 49, wherein the monitoring device (12) relays real-time flow and output volumes in the microfluidic channel (29) to determine the flow rate required in the chip cartridge (20).

51. A single-use, hermetically sealed chip cartridge (20) for use in nanoparticle synthesis or reagent manufacturing, the chip cartridge (20) comprising a base (21) and a plurality of spaced apart wells (22) there upon, wherein the wells (22) are arranged in a first row (22a), a second row (22b), and a third row (22c); wherein the well (22) in thefirst row (22a) acts as a first inlet well (24); wherein the well (22) in the second row (22b) acts as a second inlet well (26); wherein the well (22) of the third row (22c) acts as an outlet well (28); and wherein each well (22) comprises at least one microfluidic channel (29), the microfluidic channel (29) comprising a first inlet (29a), a second inlet (29b) and an outlet (29c) which are in fluid communication with the first inlet well (24), the second inlet well (26) and the outlet well (28), respectively;wherein the chip cartridge (20) is hermetically sealed by a chip cartridge cap (200).

52. The single-use, hermetically sealed chip cartridge (20) according to Claim 51, further comprising one or more inlet valves selected from an active valve or a passive valve, or a combination thereof.

53. The single-use, hermetically sealed chip cartridge (20) according to Claim 51 or 52, further comprising an outlet valve selected from an active valve or a passive valve.

54. The single-use, hermetically sealed chip cartridge (20) according to Claim 52 or Claim 53, wherein the active valve is selected from the group comprising a solenoid valve, a pneumatic membrane valve, a piezoelectric valve, and a thermal actuated valve.

55. The single-use, hermetically sealed chip cartridge (20) according to Claim 54, wherein the active valve is actuated by an external control mechanism selected from an electrical, a thermal, a magnetic, a piezoelectric or a pneumatic activator.

56. The single-use, hermetically sealed chip cartridge (20) according to Claim 52 or Claim 53, wherein the passive valve is selected from the group comprising a one-way valve, a checkvalve, a passive valve, an active valve, a duckbill valve, an umbrella valve, a ball check valve, and a flapper valve.

57. The single-use, hermetically sealed chip cartridge (20) according to Claim 56, wherein the passive valve is configured to open in response to an applied gas pressure during liquid injection and to close automatically when the internal pressure returns to ambient pressure.

58. The single-use, hermetically sealed chip cartridge (20) according to Claim 53, wherein the outlet valve is configured to close automatically after a mixing operation,such that the chip cartridge (20) remains hermetically sealed during subsequent transport or storage.

59. The single-use, hermetically sealed chip cartridge (20) according to any one of Claims 52 to 58, wherein the active valve or the passive valve, or both, are integrated in the chip cartridge cap (200).

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