Systems and methods for high-throughput processing of multiple samples

Abstract

The present invention discloses a tangential flow filtration (TFF) chip. The TFF chip features a chip substrate assembly having a closed fluid channel and a membrane assembly. The membrane assembly includes a concentrating membrane or filtration membrane, a low-volume channel, and a standard-volume channel. One or more diaphragm / valve complexes on an integrated transfer tube and valve insert are interposed along the closed fluid channel. The closed fluid channel is in valve-controlled fluid communication with the low-volume channel and the standard-volume channel. This enables efficient and controlled filtration within a compact, integrated TFF chip design.

Description

Technical Field

[0001] Cross-reference to related applications. This application claims priority to U.S. Provisional Patent Application No. 63 / 525,503, filed Jul. 7, 2023, which is hereby incorporated by reference in its entirety.

[0002] This disclosure relates to systems and methods for high-throughput automated processing of multiple samples. In particular, it relates to high-throughput automated sample concentration, high-throughput automated diafiltration (dialysis filtration), or high-throughput automated tangential flow filtration for multiple samples.

Background Art

[0003] Sample concentration, diafiltration, and tangential flow filtration (TFF) are related processes commonly used in the fields of biotechnology and biochemical engineering for the purification and concentration of biomolecules. Of course, these techniques are used in a variety of other settings and applications.

[0004] Sample concentration is a process of increasing the concentration of specific components (e.g., proteins, nucleic acids) in a solution. Sample concentration typically involves removing a solvent (usually water) from the sample. This is to reduce its volume and thereby increase the concentration of the desired components. Sample concentration can be achieved through various methods, including evaporation, centrifugation, and membrane-based techniques such as ultrafiltration or microfiltration.

[0005] Diafiltration is a membrane-based process. It is used to remove small molecules or salts from a solution while retaining larger molecules. This typically involves adding a fresh solvent (buffer) to the sample. The addition is done while simultaneously removing an equal volume of permeate through the membrane, effectively washing away the small molecules. Diafiltration can be used for buffer exchange, desalting, or removal of unwanted low-molecular-weight substances from a solution. It is often used after sample concentration to further purify concentrated samples.

[0006] TFF is a membrane filtration technique. In membrane filtration, the feed solution flows tangentially across the surface of a semipermeable membrane. This reduces membrane fouling and allows for more continuous operation. In TFF, the differential pressure across the membrane facilitates filtration, allowing smaller molecules to pass through (permeate) while retaining larger molecules (retaining solution). TFF can be used for both concentration and diafiltration processes, which typically enables large-scale batch processing of samples. TFF is a widely used technique for the separation and concentration of biological macromolecules, including proteins and nucleic acids.

[0007] TFF can be used to concentrate a sample by continuously removing the solvent while retaining larger molecules. This reduces the sample volume and increases the concentration of the target molecule. Furthermore, after the initial concentration step, diafiltration can be performed using the same TFF setup by adding fresh buffer to the feed solution and removing the permeate. This helps to remove salts, replace the buffer, or eliminate small contaminants. In particular, in one example, TFF is used to concentrate the sample first. Once concentrated, diafiltration is then performed to further purify the sample by removing unwanted small molecules. In specific examples, both steps can be performed in the same conventional TFF system by adjusting the processing parameters.

[0008] Therefore, there are conventional systems and methods that achieve one or more of the above. However, these conventional systems and methods have several challenges, shortcomings, limitations, and problems.

[0009] Spin column concentrators, such as those from Amicon, are commonly used for sample concentration in molecular biology and protein research applications. Spin column concentrators utilize centrifugal force to facilitate the concentration process. A spin column concentrator typically consists of a filtration membrane inside a plastic column. The plastic column is placed inside a centrifuge tube. Typically, the column is packed with sample up to approximately 15.0 milliliters or less. Centrifugation is then applied to remove excess solvent or buffer, resulting in a concentrated sample within the column. Unfortunately, spin column concentrators require a considerable amount of user contact time and lack process control. Furthermore, the use of spin column concentrators is not feasible for large-scale batch processing or scaling to continuous flow processing of samples.

[0010] Slide-A-Lyzer® by Thermo Fisher Scientific is a dialysis device used for buffer exchange and sample purification. It utilizes a dialysis membrane to allow small molecules and salts to diffuse out of a sample while retaining larger biomolecules. Slide-A-Lyzer® is a more passive technique, simply providing a manual and moderately scalable method for basic experimental work, in one example. This technique is suitable for smaller volumes / batch and simpler applications.

[0011] The Minimate® system by Pall is a benchtop TFF system. Benchtop TFF systems are intended for small to medium-scale sample concentration and diafiltration. In contrast to Big Tuna® and Ambr Crossflow®, in one example, the Minimate® system simply provides a benchtop solution for smaller operations and is not effective for throughput processing development or large-scale operations.

[0012] The Big Tuna® system by Unchained Labs is designed for automated buffer exchange and sample concentration. It typically utilizes dead-end filtration technology, but can be adapted to TFF. It is used for buffer exchange and concentration processes, where buffer exchange involves the removal of one buffer or solution from a sample and its replacement with another. In one example, the Big Tuna® system relies on positive pressure to pass the sample through the membrane. This is a harsh process for protein-based samples, and especially vector-based samples. Furthermore, the system relies on a liquid handler to add buffer to the sample. In another example, the Big Tuna® system lacks individual sample control, often leading to failures due to over-concentration and sample loss.

[0013] The Sartorius Ambr Crossflow® system is a parallel automated crossflow filtration system. It employs TFF for simultaneous processing of multiple samples through multiple channels. Specifically, it uses 4 to 16 TFF channels, which run in parallel. In one example, the Ambr Crossflow® system is burdened by a hold-up volume between approximately 5.0 ml and 10.0 ml, and despite being called "automatic," the user must fill each sample.

[0014] Furthermore, in most, if not all, of the examples listed, conventional systems also suffer from the problem of dead volume. Here, one or more residual samples / fluids remain trapped within the system. This leads to sample loss, contamination, and inaccurate fluid dispensing.

[0015] Thus, conventional TFF systems and methods require considerable user intervention throughout the entire process. User intervention includes manual sample loading, manual buffer changes, and / or manual cleaning. User intervention can introduce errors and unintended variability into the results. This makes the process time-consuming and prone to inconsistency. User intervention also increases the risk of contamination and / or artifacts, especially when processing multiple samples at once or consecutively. Cleaning and decontamination between sample runs may be effective, but may not always be sufficient to eliminate the risk of biological contamination caused by user intervention.

[0016] Furthermore, extending conventional TFF processing to handle multiple samples simultaneously presents significant challenges. Current systems often lack the ability to efficiently and consistently process multiple samples in parallel and sequentially. This is a bottleneck in high-throughput applications. Conventional TFF systems also often lack comprehensive and user-friendly control and monitoring capabilities. Precise control of processing parameters such as intermembrane pressure, flow rate, and volume exchange can be difficult to achieve. This impacts processing reproducibility and overall efficiency, especially when attempting to process multiple samples in parallel.

[0017] Thus, none of the existing approaches to achieving high-throughput automated processing of multiple samples using ultrafiltration or microfiltration membranes (e.g., high-throughput automated sample concentration, high-throughput automated diafiltration, and high-throughput automated tangential flow filtration of multiple samples) provide a comprehensive solution that combines the features described in this disclosure. [Overview of the project]

[0018] In some embodiments, the techniques described herein relate to tangential flow filtration (TFF) tips. TFF tips are (a) Chip substrate assembly defining a closed fluid channel, (b) A membrane assembly comprising (i) a concentrating membrane or a filtration membrane, (ii) a low-volume pathway, and (iii) a standard-volume pathway, (c) Integrated transfer tube, and (d) Includes one or more diaphragm / valve complexes on a valve insert. One or more diaphragm / valve complexes are interposed along a closed fluid passage. The closed fluid passage is in valve-controlled fluid communication with a low-capacity path and a standard-capacity path.

[0019] In some aspects, the technology described herein relates to a high-throughput automated multi-sample processing system. The system is (a) Base unit, (b) A tangential flow filtration (TFF) tip configured to operate with the base unit. (c) Automation subsystem for engaging with TFF chip, and (d) Including a housing that defines an envelope for the system. TFF chips are (i) Chip substrate assembly defining a closed fluid channel, (ii) Membrane assembly, (iii) Integrated transfer tube, and (iv) It includes one or more diaphragm / valve composites intervening along a closed fluid flow path. The membrane assembly includes (i) a concentration or filtration membrane, (ii) a low-volume path, and (iii) a standard-volume path.

[0020] In some aspects, the techniques described herein relate to a tangential flow filtration (TFF) chip. The TFF chip (a) A chip substrate assembly defining a closed fluid flow path, and (b) includes one or more integral elastomeric components. The chip substrate assembly (i) an air side, and (ii) a liquid side. The air side receives one or more pressurized air inputs, and the liquid side is configured to receive a pressurized liquid flow for the closed fluid flow path. One or more integral elastomeric components are disposed between the air side and the liquid side of the chip substrate assembly to form one or more diaphragm / valve composites. One or more diaphragm / valve composites intervene along the closed fluid flow path on the liquid side of the chip substrate assembly.

[0021] In some aspects, the techniques described herein relate to a method for achieving high-throughput automated multi-sample processing. The method (a) assembling a chip substrate assembly (b) providing a liquid flow to the closed fluid flow path of the chip substrate assembly, and (c) operating one or more diaphragm / valve composites. The step of assembling the chip substrate assembly (i) providing an air side (ii) providing a liquid side, (iii) providing one or more diaphragm / valve composites, and (iv) The process includes stacking the air side and the liquid side with one or more diaphragm / valve complexes between them. The air side is configured to accept one or more pressurized air inputs. In the stacking process, a closed fluid channel is defined between the air side and the liquid side. Furthermore, in the stacking process, one or more diaphragm / valve complexes are interposed along the closed fluid channel on the liquid side of the chip substrate assembly. [Brief explanation of the drawing]

[0022] Many aspects of this disclosure will be better understood by referring to the following drawings. The components in the drawings are not necessarily to scale. Instead, the emphasis is on clearly illustrating the principles of this disclosure. Furthermore, in the drawings, similar reference numerals indicate corresponding parts in several figures. These implementations and embodiments should be recognized as merely illustrative of the principles of this disclosure.

[0023] [Figure 1] Figure 1 is a perspective view showing an example of a high-throughput automated multi-sample processing system 1 according to this disclosure.

[0024] [Figure 2] Figure 2 is a front elevation view showing an example of a base unit having a millifluidic / microfluidic TFF chip according to this disclosure.

[0025] [Figure 3] Figure 3 is a front elevation view showing an example of a base unit without a millifluidic / microfluidic TFF chip according to this disclosure.

[0026] [Figure 4] Figure 4 is a perspective view showing an example of a base unit without a base unit housing according to this disclosure.

[0027] [Figure 5]Figure 5 is a side elevation view showing an example of a base unit without a base unit housing according to this disclosure.

[0028] [Figure 6] Figure 6 is a perspective view showing an example of a millifluidic / microfluidic TFF chip comprising an integrated membrane assembly and fluid handling elements according to this disclosure.

[0029] [Figure 7] Figure 7 is a front elevation view showing an example of a millifluidic / microfluidic TFF chip comprising an integrated, individually controllable diaphragm pump and valve according to the present disclosure.

[0030] [Figure 8] Figure 8 is an enlarged fractured perspective view showing an example of a valve insert for a millifluidic / microfluidic TFF tip at a limit point according to this disclosure.

[0031] [Figure 9] Figure 9 is a perspective view showing an example of a valve insert for a millifluidic / microfluidic TFF tip according to this disclosure.

[0032] [Figure 10] Figure 10A is a first perspective view showing an example of an automated subsystem relating to this disclosure. The automated subsystem includes a mobile gantry having a robotic gripper.

[0033] Figure 10B is a second perspective view showing an exemplary automation subsystem of Figure 10A.

[0034] [Figure 11] Figure 11 is a perspective view showing an example of an automated subsystem according to the present disclosure. The automated subsystem includes a gantry configured to move up and down. The automated subsystem also has a robotic gripper that articulates around a sliding pivot point on the gantry.

[0035] [Figure 12] Figure 12 is a perspective view showing an example of a gantry and tube rack relating to this disclosure.

[0036] [Figure 13] Figure 13 is a perspective view showing an example of one or more automated tube racks and / or one or more cassette racks / chips relating to this disclosure.

[0037] [Figure 14] Figure 14 is a perspective view showing an example of one or more automated tube racks and / or one or more cassette racks / chips relating to this disclosure.

[0038] [Figure 15] Figure 15 is a perspective view showing an example of one or more automated tube racks and / or one or more cassette racks / chips relating to this disclosure. [Modes for carrying out the invention]

[0039] The subject matter currently disclosed is described more fully below with reference to the accompanying drawings. These drawings show several embodiments of the subject matter currently disclosed, but not all embodiments. Similar numbers refer to similar elements throughout. The subject matter currently disclosed may be embodied in many different forms. The subject matter currently disclosed should not be construed as being limited to the embodiments described herein. Rather, these embodiments are provided to satisfy the applicable legal requirements of this disclosure. Indeed, many modifications and other embodiments of the subject matter currently disclosed described herein will be recalled by those skilled in the art to whom the subject matter currently relates. This is due to the benefit of the teachings presented in the foregoing description and the accompanying drawings. Therefore, it should be understood that the subject matter currently disclosed is not limited to the specific embodiments disclosed. It should also be understood that modifications and other embodiments are intended to be included within the scope of the attached claims.

[0040] Throughout this specification and the claims, the terms “comprise,” “comprises,” and “comprising” are used in a non-exclusive sense, unless the context requires otherwise. Similarly, the term “includes” and its grammatical variations are intended to be non-exclusive, such that the enumeration of items in a list does not preclude other similar items that may be substituted for or added to the listed items.

[0041] Furthermore, throughout this specification and the claims, the terms “air” and “air side” are used for convenience and simplification and not to limit the scope of this disclosure. In one embodiment, gases other than air, such as carbon dioxide, nitrogen, etc., are assumed. Accordingly, the air side is not limited to accepting only pressurized air input. In one embodiment, the air side may accept pressurized gas input. Similarly, a pneumatically operated valve may, in one embodiment, be a gas-operated valve.

[0042] "Hold-up volume" is used to refer to the volume of fluid remaining in a system after the fluid flow has stopped or after it has been purged from the system. This fluid is retained in the system due to its design and operating conditions. Hold-up volume is important in a variety of applications. In such applications, knowing the total volume of fluid retained in the system is essential for understanding the complete fluid dynamics. Relatively high hold-up volume can lead to sample loss, dilution, or contamination. This affects the accuracy and efficiency of the processing and the system. It also affects the time required for flushing and cleaning the system.

[0043] "Dead volume" is used to refer to the volume of fluid that stagnates or does not flow efficiently within a system. Dead volume of a fluid is often characterized by poor mixing or inefficient fluid movement. Dead volume is important in a variety of applications where precise control of fluid flow and mixing is required. It typically occurs in areas where fluid can be trapped, such as connectors, fittings, or corners. Relatively high dead volume can lead to problems such as incomplete reactions, cross-contamination, or delayed response times in analytical systems.

[0044] In additional context, dead-end filtration is a filtration technique. Dead-end filtration operates by passing the entire volume of a sample through a filtration membrane. In dead-end filtration, the sample is applied to one side of the filtration membrane. The filtrate (the liquid passing through the membrane) is collected on the other side. Dead-end filtration is commonly used for small-scale filtration applications, such as laboratory-scale separation and sample clarification. In contrast, TFF operates by continuously flowing the sample tangentially across the surface of a semipermeable membrane. The sample is then circulated across the membrane surface. During this time, differential pressure is applied to facilitate the filtration process. The filtrate containing undesirable molecules passes through the membrane and is collected separately. Meanwhile, the retaining liquid (the remaining sample) is left behind the membrane barrier. Thus, TFF is particularly useful for the concentration, separation, and diafiltration of large volumes of samples, such as in biopharmaceutical production and protein purification.

[0045] In further context, protein production is a complex process. This process involves several combinations of biological cells, cellular components, chemical reactions, separation methods, and the use of filtration. The final result of this process (and many intermittent steps) is typically a very dilute solution. This solution contains the target protein and low-molecular-weight substances such as salts that could not be removed in previous steps. To continue the purification process or to utilize the protein for scientific or industrial purposes, the solution typically requires concentration and buffer exchange. One of the most common methods for this is ultrafiltration.

[0046] Ultrafiltration, like standard filtration, involves a selective permeable membrane. By applying force (typically pressure) to the starting mixture on the sample side of the membrane, smaller components are allowed to pass across the membrane while proteins remain on the sample side, thereby concentrating the proteins. The same principle can be used for buffer exchange by resuspending the concentrated proteins in the desired buffer and repeating the concentration process.

[0047] If the sample flow is unidirectional and across the membrane, such a filtration setup is for dead-end filtration. While dead-end filtration is simple to manufacture and use, it has a significant drawback: it creates a concentration gradient in the membrane. This is because the mixing of the sample relies solely on diffusion and convection. The concentration gradient causes a decrease in filtration flow rate, creating conditions that, for example, impair the stability of proteins.

[0048] In contrast to dead-end filtration, the TFF systems and methods of this disclosure have a constant recirculating flow parallel to the membrane on the sample side. In one embodiment of TFF, or cross-flow filtration, also known as cross-flow filtration, both pressure and flow are typically generated using a combination of a peristaltic pump and a flow splitter. Proteins are constantly removed from the membrane surface and the solution is mixed. This ensures that the solution has a consistent concentration throughout. The absence of a concentration gradient makes TFF or cross-flow filtration gentler on proteins and provides better flow characteristics. Other advantages of TFF or cross-flow filtration include ease of online monitoring and scalability. Furthermore, the membrane can be cleaned and reused or discarded. Also, as described herein, the tubing and / or solution containers can be cleaned and rinsed after use.

[0049] A multi-channel cross-flow system, also known as a parallel flow system, is a specific configuration of a TFF system. These systems are designed to handle multiple samples simultaneously. They are particularly useful in high-throughput applications where efficiency and productivity are critical. In a multi-channel cross-flow system, multiple independent channels or flow paths are fabricated within a single TFF system, tip, or cassette. Each channel consists of its own filtration module, which includes a semipermeable membrane and associated components such as a pump, valve, and sensor. Samples are supplied to each of these channels, and the filtration process occurs simultaneously for all channels.

[0050] Furthermore, in a more detailed context, ultrafiltration membranes and microfiltration membranes are incorporated into TFF chips or cassettes according to this disclosure. This enables the selective separation and filtration of particles and molecules. Ultrafiltration membranes have a defined molecular weight cutoff (MWCO). The MWCO determines the size range of molecules that the membrane can hold or allow to pass through. These membranes typically have smaller pore sizes compared to microfiltration membranes. This enables the separation of smaller molecules. Ultrafiltration membranes are generally designed to hold macromolecules such as proteins, nucleic acids, and polysaccharides. On the other hand, microfiltration membranes are designed to allow smaller molecules, such as salts, small proteins, and solvents, to pass through.

[0051] Microfiltration membranes have larger pore sizes compared to ultrafiltration membranes. This allows for the separation of larger particles, such as cells, microorganisms, and particulate matter. These membranes are often used for clarification, sterilization filtration, and removal of debris or contaminants from liquid samples. Within the context of TFF tips, microfiltration membranes can be employed as prefilters or primary filtration media, depending on the specific application. As prefilters, they remove larger particles or aggregates, preventing their accumulation on subsequent ultrafiltration membranes. As primary filtration media, microfiltration membranes selectively retain larger particles while allowing smaller molecules and solvents to pass through. This facilitates size-based separation or purification.

[0052] With the above context in mind, systems and methods for high-throughput automated processing of multiple samples are configured for many practical applications. These applications include, but are not limited to, (1) protein production, purification, and concentration; (2) sample preparation for analytical assays; (3) buffer exchange for downstream processing; and (4) desalting.

[0053] Generally, systems for high-throughput automated processing of multiple samples typically include assemblies, subsystems, reservoirs, manifolds, fluid handling elements, and membranes. These systems also typically include precision control mechanisms to adjust the fluid flow rate (or throughput per unit time) and / or the volume of fluid being flowed. In addition, some systems typically include other common automation or robotic components, subsystems, or subassemblies.

[0054] For example, some systems include one or more base units within a housing, and one or more control units configured to operate and monitor the entire system. These control units include one or more processors that oversee the system's operation, and one or more controllers having one or more integrated firmwares. The control units also include pre-programmed protocols and one or more installed software for sample handling and automation.

[0055] Other systems include a fluid source, reservoir, or vent. These other systems also include related components, subsystems, or subassemblies, such as filter media or filtration systems, for ensuring purity and sterility and preventing contamination.

[0056] Other systems include pumps such as peristaltic pumps or syringe pumps. These are used to generate fluid flow and / or to drive fluid through the system.

[0057] Other systems include fluid channels or fluid manifolds through which fluids move from a source through the system.

[0058] Other systems include an integrated transfer tube through which fluid can be received, or returned, recirculated, or recycled.

[0059] Other systems include one or more ports / tips / nozzles / outlets. Fluids are drawn into and / or discharged into or out of the system through these ports / tips / nozzles / outlets. The configuration of these ports / tips / nozzles / outlets may vary depending on factors such as the size and geometry of the final fluid container, as well as the viscosity of the fluid.

[0060] Other systems include integrated functions for chip handling equipment. In some cases, for example, the system may be integrated with robotic or automated equipment that handles and / or tracks chips or cassettes. This facilitates high-throughput experiments.

[0061] Other systems include a user interface, through which operators can, for example, set up experiments, monitor progress, adjust parameters, enable real-time data visualization, and perform control adjustments.

[0062] Other conventional systems include a specific control mechanism, which, for example, includes a sensor system that monitors flow rate, pH, concentration, and adjusts pump speed, etc.

[0063] In the context of millifluidics and microfluidic systems, the definition of a system for high-throughput automated processing of multiple samples using ultrafiltration or microfiltration retains many of the core components and considerations outlined above and herein, but there are several nuances and additional factors to consider. For example, milli / microfluidic systems are characterized by their small size, with channels and wells typically at the milli, micro, or even nanoscale. This necessitates miniaturization of components such as pumps, valves, and fluid pathways to achieve precise control and manipulation of the fluid. However, this gives rise to a set of problems, challenges, and deficiencies of their own.

[0064] Various methods and systems have been developed to attempt to achieve efficient and effective high-throughput automated processing of multiple samples using ultrafiltration or microfiltration. However, these existing approaches have limitations and drawbacks that have hindered their effectiveness and accuracy. I. Exemplary Use Case Scenarios

[0065] The systems and methods for high-throughput automated processing of multiple samples according to this disclosure (for example, in one embodiment, about 30 to about 54 samples per run when using about 1.5 ml of sample, or in another embodiment, about 30 to about 48 samples per run when using about 15.0 ml of sample, or in another embodiment, about 24 to about 36 samples per run when using about 50.0 ml of sample) present a novel approach, as well as one or more technical steps and / or solutions, to address the challenges and shortcomings of the prior art.

[0066] For example, in one embodiment, the present disclosure provides a system and associated method for sample processing using an ultrafiltration membrane or microfiltration membrane, wherein the system is configured for: (1) automated loading and individual physical control of one or more samples; (2) parallel sample processing of two or more samples; and / or (3) use of the same membrane path for rinsing, cleaning, sterilization, and / or reuse. This enables the system and method to perform high-throughput automated processing of multiple samples with substantially no user intervention or with relatively minimal user intervention. The system also benefits from other advantages and improvements described herein.

[0067] In another embodiment, the systems and methods relating to the present disclosure provide miniaturized, or "smaller-scale," ultrafiltration / microfiltration systems on-chip. These systems on-chip are defined by relatively low hold-up volume and relatively low dead volume (compared to the prior art) and can be used (alone or together) in automated, laboratory-sized machines or mechanisms.

[0068] In another embodiment, the systems and methods relating to the present disclosure provide disposable (e.g., consumable) assemblies, tips, or cassettes for sample concentration, diafiltration, or tangential flow filtration of multiple samples. Alternatively, preferably, the systems and methods provide non-disposable (e.g., reusable) assemblies, tips, or cassettes for sample concentration, diafiltration, and tangential flow filtration of multiple samples.

[0069] In another embodiment, the systems and methods relating to the present disclosure provide reusable and / or disposable TFF chips or cassettes that are capable of a wide range of practical applications and implementations. Such applications and implementations include, for example, the filtration or purification of proteins in quantities of about 1.0 kilodalton to about 500.0 kilodaltons, the filtration or purification of lipid nanoparticles in quantities of about 50.0 nanometers to about 1000.0 nanometers or about 1.0 micrometer, the filtration or purification of viruses in quantities of about 25.0 nanometers to about 500.0 nanometers, the filtration or purification of nucleic acids in quantities of about 1.0 kilodalton to about 1000.0 kilodaltons or about 1.0 megadaltons, and the filtration or purification of cells in quantities of about 1.0 micrometer to about 30.0 micrometers.

[0070] In another embodiment, the systems and methods relating to this disclosure provide reusable and / or disposable TFF chips or cassettes for: optimizing protein preparation workflows by enabling rapid and gentle processing, including efficient processing, concentration, formulation, desalting, and refolding of lipid nanoparticles, liposomes, and polymer nanoparticles for drug delivery and therapeutic effects; ensuring gentle and efficient removal of unbound small molecules from various crude biomolecular labeling reactions; simplifying in vitro synthesis of RNA, as well as linear or plasmid DNA, by efficient concentration and buffer exchange; streamlining rapid recovery of cells, extracellular vesicles, enzymes, and virus-like particles (VLPs) while ensuring high product yield and quality; gently concentrating and buffer exchanging adeno-associated virus vectors (AAVs), bacteriophages, and lentiviruses while preserving their structure for effective applications; optimizing the formulation of DNA, RNA, and polysaccharide vaccines to obtain optimal, stable, and cost-effective results; and many other applications.

[0071] In another embodiment, the systems and methods relating to the present disclosure provide a TFF chip or cassette with an integrated fluid handling element, such as a pump, valve, fluid path, fluid flow path, air path, port, flow path, return line, limit point, and transfer tube.

[0072] In another embodiment, the systems and methods relating to the present disclosure provide reusable and / or disposable TFF chips or cassettes comprising multiple flow paths or fluid paths, as well as multiple membrane regions or assemblies having membranes and permeation paths. These can be run in parallel or separately to achieve low final output / sample volume, low dead volume results, and / or low hold-up volume without impairing efficient or effective concentration or filtration rates.

[0073] In another embodiment, the systems and methods relating to the present disclosure provide a TFF chip or cassette comprising a plurality of membrane assemblies ranging from about 3.5 square centimeters to about 12 square centimeters, where the membrane material and MWCO size are defined as follows: modified polyethersulfone (mPES) 5, 10, 30, 50, 100, and 300 kilodaltons, regenerated cellulose (RC) 5, 10, 30, and 100 kilodaltons.

[0074] In another embodiment, the systems and methods according to the present disclosure provide a TFF chip or cassette comprising one or more membrane assemblies. Each membrane assembly comprises a hollow fiber, flat membrane, or spirally wound concentrating or filtering membrane capable of selectively separating molecules based on size and other properties. In another embodiment, the one or more membrane assemblies are configured to run in parallel or separately. Furthermore, in another embodiment, the membrane assembly includes multiple flow paths for size exclusion using multiple cutoffs and multiple exposed membrane regions. All of these are located within a single chip or cassette. For example, in another embodiment, high and low molecular weight cutoffs are incorporated into the same chip or cassette to create a filter band. Here, an initial sample can be purified to remove small impurities, and then a filter with a larger molecular weight cutoff can be applied to, for example, sterilize each sample or remove impurities larger than the target biological substance.

[0075] In another embodiment, the systems and methods of the present disclosure provide a TFF chip configured to be housed in a base unit of a broader high-throughput automated multi-sample processing system. In another embodiment, the TFF chip comprises a fluid handling element and a membrane assembly. In another embodiment, the base unit includes one or more liquid sources for the chip (e.g., one or more sample sources, one or more buffer sources, one or more reagent sources, etc.) and an air / gas source for clearing / cleaning / drying the chip and / or for operating one or more diaphragm pumps and valves of, for example, the fluid handling element on the chip.

[0076] In another embodiment, the systems and methods relating to the present disclosure provide a TFF chip having multiple layers. In another embodiment, each of the multiple layers defines air passages, fluid passages, and ports for airflow control and liquid flow control. Furthermore, in another embodiment, when these layers are stacked, engaged, and assembled, fluid handling elements and one or more membrane assemblies are located between the outer layers of the multiple layers.

[0077] In another embodiment, the system and method according to the present disclosure provides a TFF chip having a plurality of layers, including four laser-bonded polystyrene layers, where one or more diaphragm pumps and membrane assemblies are positioned between two intermediate layers of the plurality of layers. In another embodiment, the layers include interference shapes, which form a seal that allows air to actuate the diaphragm pump and valves and the like, and allows liquid to move around the pump operating chip and over the membrane.

[0078] In another embodiment, the systems and methods of the present disclosure provide a TFF chip configured to be retained and stored in a membrane storage region until needed, the membrane storage region holding a membrane storage solution (e.g., about 20% glycerol and about 0.02% sodium azide).

[0079] In another embodiment, the systems and methods relating to this disclosure provide a TFF tip that also enables fluid channeling (induction) and discharge.

[0080] In another embodiment, the systems and methods relating to this disclosure provide a TFF tip that also enables fluid channeling, recirculation / recovery, and discharge.

[0081] In another embodiment, the systems and methods relating to the present disclosure provide a TFF tip configured to draw liquid from a container or reservoir and to return the treated or untreated liquid to the container or reservoir. All of these are configured on the tip as part of an integrated transfer tube.

[0082] In another embodiment, the systems and methods relating to the present disclosure provide a TFF tip having a pair of tubes configured to extend into a container, tube, or reservoir. One of the two tubes is used to draw liquid from the container, tube, or reservoir, and the other of the two tubes is used to return liquid to the container, tube, or reservoir.

[0083] In another embodiment, the systems and methods relating to this disclosure provide a TFF chip or cassette that can function with only a relatively small sample volume, for example, from about 250.0 nanoliters to about 100.00 milliliters.

[0084] In another embodiment, the systems and methods relating to the present disclosure provide a TFF chip or cassette having a relatively low hold-up volume, for example, from about 190.0 microliters to about 250.0 microliters.

[0085] In another embodiment, the systems and methods relating to the present disclosure provide a TFF chip or cassette that operates with a relatively low dead volume, such as between about 10.0 microliters and about 20.0 microliters.

[0086] In another embodiment, the systems and methods relating to the present disclosure provide a TFF tip or cassette with an improved back pressure adjustment function, having an integrated valve / pump for continuous flow, continuous supply pressure pumping, and enabling intermembrane pressures ranging from about 0.1 pounds per square inch to about 32.0 pounds per square inch.

[0087] In another embodiment, the systems and methods relating to the present disclosure provide TFF chips or cassettes that can be loaded and unloaded from various system components and / or stations via an automated subsystem (e.g., via one or more robotic grippers).

[0088] In another embodiment, the systems and methods relating to this disclosure include one or more unidirectional valves and one or more bidirectional valves.

[0089] In another embodiment, the systems and methods relating to this disclosure include a fluid valve or microfluidic valve configured as a diaphragm / valve complex.

[0090] In another embodiment, the systems and methods relating to the present disclosure include a diaphragm / valve complex configured as two or more separate parallel valve clusters.

[0091] In another embodiment, the systems and methods relating to the present disclosure include one or more valve clusters, each configured as one or more separate single elastomer components. Each elastomer component has a separate region defining one or more valves and / or one or more diaphragm pumps.

[0092] In another embodiment, the systems and methods relating to the present disclosure include diaphragm / valve complexes having differently configured valve clusters. For example, one or more valve clusters may have a small diaphragm pump and another valve cluster may have a large diaphragm pump, or one diaphragm pump may have certain functions and connections and another diaphragm pump may have different functions and connections.

[0093] In another embodiment, the systems and methods relating to this disclosure include a diaphragm / valve complex configured as two or more valve clusters.

[0094] In another embodiment, the systems and methods relating to the present disclosure include a single, integrated elastomer component, or a plurality of valve clusters integrated with a valve insert. The elastomer component has separate regions that define one or more valves and / or one or more diaphragm pumps.

[0095] In another embodiment, the systems and methods relating to the present disclosure include, as part of a diaphragm / valve complex, a single elastomer component having a double-acting diaphragm pump, or a valve insert.

[0096] In another embodiment, the systems and methods relating to the present disclosure include a diaphragm / valve complex having a plurality of separate membrane channels and transfer tubes and a double-acting diaphragm pump that is in fluid communication under valve control.

[0097] In another embodiment, the systems and methods relating to the present disclosure include a plurality of valve clusters, each having one diaphragm pump and three individually controllable elastomer valves.

[0098] In another embodiment, the systems and methods relating to this disclosure provide a low-shear force flow that acts gently on biological agents and proteins.

[0099] In another embodiment, the systems and methods described herein provide chip assemblies with good sealing performance and long life / durability.

[0100] In another embodiment, the systems and methods relating to this disclosure include diaphragm pumps having a top hat configuration, a multilevel configuration, or a dome-shaped diaphragm configuration.

[0101] In another embodiment, the systems and methods relating to this disclosure provide a function for priming one or more chips.

[0102] In another embodiment, the systems and methods relating to this disclosure provide a function for recovering liquid from a chip.

[0103] In another embodiment, the systems and methods relating to the present disclosure provide a function for recirculating a flow through any number of paths among one or more fluid channels.

[0104] In another embodiment, the systems and methods relating to the present disclosure provide a function for drawing a substance out of and through a transfer tube and pushing it upstream.

[0105] In another embodiment, the systems and methods relating to this disclosure provide a high-throughput automated multi-sample processing system having compact dimensions (e.g., approximately 830.0 mm × approximately 810.0 mm × approximately 700.0 mm). This allows for use on a desktop or workbench, and also enables the system to be used in various biosafety cabinets to accommodate workflows requiring ventilation or isolation, for example.

[0106] In another embodiment, the system and method of the present disclosure provide a high-throughput automated multi-sample processing system capable of processing up to 54 multi-samples per run, and equipped with independently configurable pressure control, and capable of running up to 5 samples in parallel.

[0107] In another embodiment, the systems and methods relating to this disclosure are configured for initial sample volumes between approximately 1.5 milliliters and approximately 100.0 milliliters, or between approximately 1.0 milliliter and approximately 150.0 milliliters, and provide a high-throughput automated multi-sample processing system that yields a low final volume of 250 microliters with a resolution of plus or minus approximately 25.0 microliters at a recommended maximum sample viscosity of approximately 100 centipores.

[0108] In another embodiment, the system and method relating to this disclosure provide a high-throughput automated multi-sample processing system configured to allow a user to set decks and processing parameters, import an upstream dispensing list, and run a protocol. The system performs this method and notifies the user upon completion. It is structured in this way.

[0109] In another embodiment, the systems and methods relating to the present disclosure provide a high-throughput automated multi-sample processing system including a valve insert having a double-acting diaphragm pump. This enables nearly continuous concentration processing and pressure gradients (reducing stress on the sample), and the permeate flow rate is up to approximately 1.7 times faster than conventional systems and up to approximately 4.0 times faster than centrifugal dead-end filters.

[0110] In another embodiment, the systems and methods relating to the present disclosure provide a high-throughput automated multi-sample processing system that enables easy cleaning between chip uses without disassembly.

[0111] In another embodiment, the systems and methods relating to the present disclosure provide an automated cleaning cycle for cleaning an entire fluid flow path without user intervention. This is a forward or reverse cleaning cycle, for example, which allows the liquid to flow backward in a direction that gradually increases the flow path size for thorough, clog-free cleaning.

[0112] In another embodiment, the systems and methods relating to the present disclosure provide a high-throughput automated multi-sample processing system configured to autonomously clean the entire liquid pathway after each run and to autonomously rinse it with, for example, a buffer or water.

[0113] In another embodiment, the systems and methods of the present disclosure provide a high-throughput automated multi-sample processing system configured to use a plurality (e.g., 4 to 8) external buffer reservoirs or on-deck buffers (e.g., 42 different buffer types). Here, buffer exchange can be performed on a volume-based or conductivity-based basis using an in-line conductivity probe.

[0114] In another embodiment, the system and method relating to this disclosure provide a high-throughput automated multi-sample processing system configured to perform permeate collection outside the base unit, with an overflow monitoring function.

[0115] In another embodiment, the system and method relating to the present disclosure provide a high-throughput automated multi-sample processing system configured to monitor the conductivity of a permeate in the range of about 1.0 to about 60.0 millisiemens per square centimeter.

[0116] In another embodiment, the system and method relating to this disclosure provide a high-throughput automated multi-sample processing system that integrates an on-deck sample cooling function capable of cooling to, for example, about 4.0°C.

[0117] In another embodiment, the systems and methods according to the disclosure include one or possibly more integral elastomer components compressed between two rigid components. This is such that open flow paths, ports, or openings on each rigid component are sealed by the elastomer or mated together to form sealed fluid flow paths. In either case, these form the air-side and liquid-side foundations of the TFF tip, where the liquid-side is configured to receive a pressurized fluid flow and the air-side is configured to receive pneumatic input to control / operate the integral elastomer components. In particular, in another embodiment, each integral elastomer component, together with the rigid components, creates a structure on the liquid side of the TFF tip that functions, for example, as three individually controlled elastomer valves and one individually controlled diaphragm pump with fluid access. Furthermore, in another embodiment, all valves function to block access to a central chamber, with a diaphragm membrane acting on its central volume. Thus, in another embodiment, the valves regulate access to input paths, recovery / recirculation paths, and distribution paths.

[0118] In another embodiment, the system and method relating to this disclosure include a chip that includes the following: (a) A chip substrate assembly that defines a sealed fluid channel, the chip substrate assembly comprising the following: (i) Air side that accepts one or more pressurized air inputs. (ii) A liquid side configured to receive a liquid flow for a sealed fluid channel. (b) One or more diaphragm / valve complexes interposed along a sealed fluid channel on the liquid side of the chip substrate assembly.

[0119] In another embodiment, the systems and methods relating to the present disclosure include a chip, where one or more diaphragm / valve complexes each include two adjacent valve clusters, each of which includes a diaphragm pump and at least one pneumatically operated and individually controllable elastomer valve.

[0120] In another embodiment, the system and method relating to the present disclosure include a tip, where each of the two adjacent valve clusters is configured as a single, integrated elastomer component located between the air side and the liquid side of the tip.

[0121] In another embodiment, the systems and methods relating to the present disclosure include a chip, where the chip substrate assembly includes an upper substrate layer and a lower substrate layer, the upper substrate layer corresponding to the air side and the lower substrate layer corresponding to the liquid side. The chip substrate assembly defines a sealed fluid channel between them.

[0122] In another embodiment, the systems and methods relating to the present disclosure include a chip. The chip substrate assembly includes an upper substrate layer, one or more substrate layers, and a lower substrate layer, the upper substrate layer corresponding to the air side and the lower substrate layer corresponding to the liquid side. The chip substrate assembly defines a sealed fluid channel between them.

[0123] In another embodiment, the systems and methods relating to the present disclosure include a chip. The sealed fluid channel is defined at least partially by an intermediate substrate layer and a fluid recess on the surface of a lower substrate layer or another intermediate substrate layer.

[0124] In another embodiment, the systems and methods relating to this disclosure include a chip. The upper substrate layer includes a port configured to receive one or more pressurized air inputs.

[0125] In another embodiment, the systems and methods relating to the present disclosure include a chip. The chip includes two or more diaphragm / valve complexes. Each of the two or more diaphragm / valve complexes constitutes a single integrated elastomer component between the air side and the liquid side of the chip substrate assembly. This forms a plurality of valve clusters interposed along a sealed fluid channel on the liquid side of the chip substrate assembly.

[0126] In another embodiment, the systems and methods relating to this disclosure include methods for achieving TFF on a chip. Such methods include: (a) The process of assembling the chip substrate assembly. Here, the process of assembling the chip substrate assembly includes the following steps: (i) A step of providing an air side configured to accept one or more pressurized air inputs. (ii) A step of providing the liquid side. (iii) A step of providing one or more diaphragm / valve composites. (iv) A step of stacking the air side and the liquid side with one or more diaphragm / valve complexes sandwiched between the air side and the liquid side. The stacking is carried out so as to define a sealed fluid channel between the air side and the liquid side. The stacking is carried out so that one or more diaphragm / valve complexes are interposed along the sealed fluid channel on the liquid side of the chip substrate assembly. (b) A step of supplying liquid flow to a sealed fluid channel of a chip substrate assembly. (c) The process of acting one or more diaphragm / valve complexes.

[0127] In another embodiment, the systems and methods relating to the present disclosure include the following methods, in which one or more diaphragm / valve complexes each include two adjacent valve clusters, and the step of acting one or more diaphragm / valve complexes includes the step of acting two adjacent valve clusters.

[0128] In another embodiment, the systems and methods relating to the present disclosure include the following methods, in which the chip comprises two or more diaphragm / valve complexes. Each of the two or more diaphragm / valve complexes is configured as a single integrated elastomer component between the air side and the liquid side of the chip substrate assembly. This forms a plurality of valve clusters interposed along a sealed fluid channel on the liquid side of the chip substrate assembly. The step of acting one or more diaphragm / valve complexes includes the step of acting the plurality of valve clusters. II. Systems and Methods

[0129] In one embodiment, the disclosure provides a high-throughput automated multi-sample processing system comprising a millifluidic or microfluidic sample processing chip having one or more diaphragm pumps. In another embodiment, one or more diaphragm pumps enable priming of fluid channels, such as buffers, within the chip. In another embodiment, diaphragm pumps enable aspirating, metering, and dispensing individual volumes of components / samples through transfer tubes, fluid channels, and / or fluid paths, for example, through membrane assembly paths or through integrated transfer tubes or outward. In another embodiment, one or more diaphragm pumps may be dome-shaped portions larger than valve components.

[0130] In one embodiment, together with a diaphragm pump, the tip includes a sealed fluid channel that serves as a passage for flow between the various components of the tip, as well as for sample introduction, holding fluid circulation, and filtrate recovery.

[0131] In one embodiment, along with one or more diaphragm pumps and one or more fluid passages, the tip also includes a valve. The valve provides, for example, opening and closing a fluid passage leading to or from a diaphragm pump, or opening and closing a fluid passage leading to or from a transfer tube, or opening and closing a fluid passage leading to or from a membrane assembly and one or more membranes.

[0132] In one embodiment, the diaphragm pumps and / or valves are grouped and arranged or formed as a diaphragm / valve complex. In another embodiment, the diaphragm pumps and / or valves are grouped and arranged or formed as a valve cluster. In another embodiment, the tip includes multiple diaphragm / valve complexes and / or valve clusters. In another embodiment, two or more valve clusters are arranged or configured as a diaphragm / valve complex. In another embodiment, the diaphragm pumps and / or valves are formed together as an integral elastomer part or component. In another embodiment, each diaphragm pump and / or valve is controlled or actuated separately and individually.

[0133] In one embodiment, one or more diaphragm pumps and valves are controlled by any suitable method, for example, a pressure control approach. In another embodiment, the diaphragm pumps and / or valves are actuated by a pneumatic or hydraulic method, and thus the tip includes a pressure port, where the pressure in the port actsuates the diaphragm of the diaphragm pump and / or valve, thereby causing the diaphragm pump and / or valve to open, close, or pump. The disclosure is not limited in this respect, and other suitable operating arrangements may be employed.

[0134] In one embodiment, for example, the present disclosure provides a diaphragm pump and / or valve controlled through a pressure inlet. The pressure inlet functions to press air against a flexible membrane, thereby closing the diaphragm pump and / or valve, or to refrain from applying pressure to the flexible membrane, thereby opening the diaphragm pump and / or valve. In another embodiment, a vacuum is applied to further facilitate the opening of the diaphragm pump and / or valve, thereby allowing for improved flow through the diaphragm pump and / or valve. In yet another embodiment, the diaphragm pump and / or valve is configured such that the application of pressure through the inlet to the flexible membrane functions to open the diaphragm pump and / or valve, and the non-application of pressure or the application of a vacuum through the inlet functions to close the diaphragm pump and / or valve. In yet another embodiment, the operation of the diaphragm / valve complex and / or valve cluster includes, for example, pumping by one or more diaphragm pumps (any number or combination) which may be repeated as desired.

[0135] It should be understood that diaphragm pumps and / or valves can be formed from a wide variety of suitable materials. Therefore, in one embodiment, diaphragm pumps and / or valves are made from elastomer materials such as silicone, rubber, polyurethane, polydimethylsiloxane, polytetrafluoroethylene (PFE), or any suitable equivalent thereto, or a suitable combination thereof. In another embodiment, diaphragm pumps and / or valves are made from suitable rigid materials such as metal or ceramic, which can be actuated through any suitable arrangement, regardless of their electrical or mechanical properties. When using rigid materials, in another embodiment, an openable hinge or gateway is employed.

[0136] In one embodiment, the diaphragm / valve complex and / or valve cluster may be formed from any suitable material and in any suitable arrangement or combination of the materials described above with respect to the diaphragm pump and / or valve. For example, in another embodiment, the diaphragm / valve complex and / or valve cluster may be molded together as a single integrated elastomer part or component, or as divided elastomer parts or components, or in any combination.

[0137] It should be understood that many alternative embodiments exist for diaphragm / valve complexes and / or valve clusters. Thus, in one embodiment, a cluster of diaphragm pumps and / or valves is used to control a specific pumping region of the chip. In another embodiment, the cluster of diaphragm pumps and / or valves is in fluid communication with only a portion of the fluid passages of the chip. In yet another embodiment, the cluster of diaphragm pumps and / or valves is in fluid communication with numerous fluid passages of the chip.

[0138] In one embodiment, the disclosure provides a high-throughput automated multi-sample processing system having a chip or cassette to be received in a base unit. The base unit has an air / gas supply subsystem adapted for air / gas flow rate control, and a fluid / liquid subsystem or supply source.

[0139] In one embodiment, the disclosure provides a high-throughput automated multi-sample processing system having a base unit. The base unit includes a base unit body, an air supply subsystem, a liquid subsystem, a clamp assembly for rigidly and firmly engaging a tip with the air supply subsystem (wherein the clamp is entirely optional), one or more high-precision load cells, and any other necessary systems, subsystems, or assemblies, such as one or more volume sensors, a waste slide, a waste container, a pressure pump and a vacuum pump, electronic equipment, a touchscreen for setting parameters for operation, and so on.

[0140] In one embodiment, the disclosure provides an air / gas supply subsystem or source. The air / gas supply subsystem or source includes an air supply unit and a bank of solenoids. The bank of solenoids is adapted, for example, to apply positive or negative pressure to a diaphragm pump or valve through the air supply unit. In another embodiment, the air / gas supply subsystem is also configured to blow air through an air passage in a tip.

[0141] In one embodiment, the disclosure also provides an outlet which may take any form, and is not limited thereto. Thus, in one embodiment, the outlet is configured as a tube or fluid channel leading to another area or part of a broader system which may include the outlet itself.

[0142] In one embodiment, the disclosure provides a fluid system including a base unit supporting a chip. In another embodiment, a system housing encloses system components and provides protection and covering to the system components, and also provides one or more structures / assemblies / subsystems for automation.

[0143] In one embodiment, the disclosure provides a fluid system including a tip manipulator, a tube / sample manipulator, and a cleaning subsystem. In another embodiment, the tip manipulator and the tube / sample manipulator are configured to appropriately manipulate or orient items or components of a system, for example, within the boundaries of the system (e.g., within the boundaries of the system housing). In another embodiment, the tip manipulator is configured to translate or orient one or more tips in horizontal (y-axis), vertical (Z-axis), and horizontal (X-axis) directions. It will be understood that the manipulator may be configured to move in any suitable manner so that, for example, one or more tips can be appropriately positioned. In another embodiment, one of the manipulators is configured to translate in any direction while the other one or more manipulators are configured to remain stationary. In another embodiment, one of the manipulators is configured to translate and / or articulate individually in any suitable direction or manner.

[0144] In one embodiment, the disclosure provides a fluid system that can be easily integrated with other robotic automation systems. The fluid system has a visual scanning subsystem useful for barcodes and QR codes (registered trademarks), as well as short-range and wireless communication subsystems for RFID read-and-write tags. In another embodiment, the system also includes one or more automated interfaces, such as electrical or pneumatic connectors or communication ports, which enable seamless integration with other automation systems. In another embodiment, the automated interfaces enable remote control, data exchange, and coordination with other components of any automation system or subsystem.

[0145] In one embodiment, the disclosure provides a fluid system to facilitate system automation, comprising a high-precision load cell, one or more gantry grippers, and corresponding one or more vision sensors, one or more proximity sensors, one or more pressure sensors, etc.

[0146] In one embodiment, the disclosure provides a fluid system that incorporates an integrated sensor, such as a pressure sensor, a flow sensor, a temperature sensor, a conductivity sensor, a pH sensor, and / or an absorbance sensor (e.g., a 280 nm sensor) (in any combination), to monitor and control key processing parameters. In another embodiment, these sensors provide real-time feedback, enabling automatic adjustment and optimization of sample processing parameters.

[0147] In one embodiment, the disclosure also provides a fluid system comprising a subsystem for cleaning, rinsing, and / or sterilizing fluid handling elements of one or more tips. In another embodiment, a diaphragm pump and valves are appropriately operated to appropriately flow or drive a cleaning fluid.

[0148] In one embodiment, the base unit includes a clamp assembly for holding the tip, an air supply assembly, and a fluid reservoir assembly. In addition, the TFF system may include a volume sensor for measuring the liquid level in the fluid reservoir assembly, a waste slide adapted to guide the permeate, and a waste container connected to the waste slide and adapted to collect the guided permeate.

[0149] In one embodiment, the base unit includes a tip / cassette holder and a clamp handle. The tip holder has slots for aligning tips. The clamp handle is adapted to push the tip holder and tips into a clamp assembly. The pushing is done so that the tips connect to and are sealed with the air supply assembly and the fluid reservoir assembly.

[0150] It should be understood that the cleaning fluid used for forward and / or reverse cleaning may include one or more combinations of various materials. These materials may include, for example, air and / or water. If the components to be dispensed are not adequately cleaned with air and / or water, a different type of purging component may be used. These purging components may include acetone, ethanol, nitrogen, carbon dioxide, or other suitable gaseous or fluid components, or combinations thereof.

[0151] Furthermore, since this disclosure is not limited thereto, it should be understood that any appropriate combination of the above embodiments can be adopted. Also, any or all of the above embodiments can be adopted in a fluid system for processing a sample. However, this disclosure is not limited thereto, and the embodiments can be used with any fluid flow path system. Various embodiments and models of this disclosure will be described in more detail thereafter with respect to the accompanying drawings. However, this disclosure is not limited to the embodiments and models shown. III. Reference to Drawings

[0152] U.S. Patents 8,016,260, 8,100,293, 8,205,856, and 8,550,298 have been assigned to the applicant of this application and are incorporated herein by reference in their entirety. Furthermore, U.S. Patent Application Publication 2021 / 0060491 has been assigned to the applicant of this application and is incorporated herein by reference in its entirety.

[0153] Referring here to the drawings, Figure 1 is a perspective view showing an example of a high-throughput automated multi-sample processing system 1 according to the present disclosure. In one embodiment, as shown in Figure 1, the microfluidic dispensing system 1 includes a housing 10 surrounding a base unit 100, i.e., base unit 100a (not shown), base unit 100b, base unit 100c, base unit 100d, and base unit 100e. In one embodiment, base unit 100d provides support, sample liquid, air, and various other necessities to a millifluidic / microfluidic TFF tip 1000d according to the present disclosure (best shown in Figures 4 and 5). The housing 10 also surrounds an automated subsystem 20 for translating, orienting, and / or manipulating various system 1 components, such as the tip 1000, one or more liquid sources 50 (e.g., one or more sample sources of different sizes, one or more buffer sources, one or more reagent sources), and one or more liquid containers 70.

[0154] More specifically, in one embodiment, as shown in Figure 1, the housing 10 encloses: an automation subsystem 20 having a movable gantry 22 configured to move forward and backward and a robotic gripper 24 configured to move left and right; a tube rack 40a located in an automation bay 30a corresponding to one or more liquid sources 50; a tube rack 40b located in an automation bay 30b; a tube rack 40c located in an automation bay 30c corresponding to one or more liquid containers 70; and a chip / cassette rack 90 located in an automation bay 30d. In one embodiment, the automation subsystem 20 is configured to move above the enclosure of the housing 10, including moving above the base unit 100, the automation bays 30, and the one or more chip / cassette racks 90. This is done so that the movable gantry 22 is movable within Cartesian space, thereby allowing the robotic gripper 24 to reach, access, and manipulate one or more liquid sources 50, one or more liquid containers 70, and / or chips 1000. Furthermore, this is done so that the automated bay 30 can assist in demarcating areas containing one or more important sample sources, buffer sources, or reagent sources of different sizes (e.g., SBS-sized containers, 1.5 mL, 15 mL, and 50 mL conical tubes).

[0155] Furthermore, in one embodiment, the movable gantry 22 includes framing, steppers, belt drive (3-axis Cartesian), linear rail axis, and any other components, assemblies, or subsystems required for the movable gantry. Furthermore, in one embodiment, the movable gantry 22 may be made of aluminum, plastic, and / or stainless steel. However, in other embodiments, the movable gantry 22 may be of any material and / or any dimensions not inconsistent with the purposes of this disclosure.

[0156] Furthermore, in one embodiment, the robot gripper 24 includes one or more V-shaped fixed fingers, a movable finger of a flat metal rod, a servo with linkage, one or more horizontal collision sensing plates, fasteners, and any other components, assemblies, or subsystems required for the robot gripper. Furthermore, in one embodiment, the robot gripper 24 may be made of aluminum, plastic, and / or stainless steel. However, in other embodiments, the robot gripper 24 may be of any material and / or any dimensions not inconsistent with the purposes of this disclosure. Furthermore, in one embodiment, the robot gripper 24 is configured to grip cylindrical objects with a diameter from about 8.0 mm to about 32.0 mm, and to grip flat objects, and to sense collisions with or without the gripped object.

[0157] Furthermore, in one embodiment, one or more tube racks 40 can be made of aluminum, acrylic, and / or stainless steel. However, in other embodiments, one or more tube racks 40 can be of any material and / or any dimensions not inconsistent with the purposes of the present disclosure. Furthermore, in one embodiment, one or more tube racks 40 are configured to hold one or more liquid sources 50 and / or one or more liquid containers 70 in a vertical position. The holding is provided with a spacing that allows a robotic gripper 24 to engage with one or more liquid sources 50 and / or one or more liquid containers 70 from above.

[0158] Furthermore, in one embodiment, the tip / cassette rack 90 can be made of aluminum, acrylic, and / or stainless steel. However, in other embodiments, the tip / cassette rack 90 can be made of any material and / or dimensions that are not inconsistent with the purposes of this disclosure. Furthermore, in one embodiment, the tip / cassette rack 90 is configured to hold the tip 1000 in a vertical position. The holding is configured to leave a gap that allows the robotic gripper 24 to engage with the tip 1000 from above.

[0159] Other embodiments of the automated subsystem 20, including a mobile gantry 22 with a robotic gripper 24, are shown in Figures 10-11. Other embodiments of the gantry 22 and one or more tube racks are shown in Figure 12. Other embodiments of one or more automated tube racks 40 and / or one or more chip / cassette racks 90 are shown in Figures 13-15. Furthermore, in one embodiment, the systems shown in Figures 13-15 are not merely automated tube racks and / or chip racks. These are entire systems in which concentration / buffer exchange takes place at some index point along a rotating carousel.

[0160] Figure 2 is a front elevation view showing an example of a base unit comprising a millifluidic / microfluidic TFF chip according to the present disclosure. In one embodiment, as shown in Figure 2, the base unit 200 includes a base unit housing 210, a liquid subsystem 230, and an air supply subsystem (not shown, best shown in Figure 3). The liquid subsystem 230 has a 15.0 ml sample source 231 on a first tube holding assembly 232, a 1.5 ml container / supply source 233 on a second tube holding assembly 234, a 50.0 ml buffer / reagent source 235 on a third tube holding assembly 236, and a corresponding high-precision load cell sensor (not shown). The air supply subsystem is adapted for airflow and pneumatic control to the chip 2000. The air supply subsystem is adapted for cleaning / purifying / drying the fluid handling element 2002 on the chip 2000. The air supply subsystem is adapted (best shown in Figures 6 and 7) to actuate the fluid handling elements on the chip 2000.

[0161] Figure 3 is a front elevation view showing an example of a base unit without a millifluidic / microfluidic TFF tip according to the present disclosure. In one embodiment, as shown in Figure 3, the base unit 300 includes a base unit housing 310, a liquid subsystem 330, and an air supply subsystem 350. The liquid subsystem 330 has a first tube holding assembly 332, a second tube holding assembly 334, and a third tube holding assembly 336. The air supply subsystem 350 provides a filtered air source for washing / cleaning / drying the tip and / or for operating one or more diaphragm pumps and valves (not shown, best shown in Figures 6 and 7) of the tip's fluid handling elements. In particular, in one embodiment, the air supply subsystem 350 includes an air source and driver / motor / pump (not shown), a port 352 (input or output), and an air passage 354. The port 352 is in fluid communication with the air source and driver for airflow and pneumatic pressure to one or more diaphragm pumps and valves of the tip. The air passage 354 is in fluid communication with an air source and driver to clean / purify / dry the fluid paths and membrane assembly paths (not shown, best illustrated in Figure 6) on the chip.

[0162] More specifically, in one embodiment, as shown in Figure 3, port 352 aligns with and corresponds to ports on the chip. In this way, when the chip engages with the base unit, port 352 also engages with the corresponding port on the chip. Each engagement is for receiving airflow and air pressure from the air supply subsystem 350 of the base unit 300.

[0163] Furthermore, in another embodiment, as shown in Figure 3, the air passage 354 includes an air passage inlet 356 and a purge valve outlet 358. The purge valve outlet 358 is connected to a waste tube of a cleaning subsystem (not shown). In this way, when the tip is engaged with the base unit, a purge valve (not shown, best shown in Figure 7) is used to blow air through the fluid handling element of the tip to the purge valve outlet 358. This is done to remove as much residual liquid as possible from the tip (as described herein).

[0164] Figure 4 is a perspective view showing an example of a base unit without a base unit housing according to the present disclosure. In one embodiment, as shown in Figure 4, the base unit 400 includes a base unit body 420 and a liquid subsystem 430. The liquid subsystem 430 has a second tube holding assembly 434 with a corresponding tare subsystem 470a, and first tube holding assemblies 432 and third tube holding assemblies 436 with corresponding tare subsystems 470b. In particular, in one embodiment, the first tube holding assembly 432, or the second tube holding assembly 434, or the third tube holding assembly 436 includes one or more tube adapters 437, a tube support 438, and a stepper motor 490. One or more tube adapters 437 are for accommodating different tube sizes (e.g., 1.5 ml conical tubes, 15.0 ml conical tubes, 1.5 ml conical tubes). The tube support 438 is for receiving and holding the appropriate tube adapter 437 for the protocol. The stepper motor 490 is used to move the tube holding assembly as needed. This allows, for example, the integrated transfer tube for the tip to be raised or lowered until it touches, for example, the bottom of the tube. This is detected by a load cell sensor, and the tube can then be lowered by a set amount.

[0165] Furthermore, in one embodiment, as shown in Figure 4, the tare subsystem 470a or tare subsystem 470b includes one or more load cell sensors (not shown), one or more linear rails 472, one or more carriages 474, and a stepper motor 490. The stepper motor 490 raises and lowers the linear rails 472 based on signals from the load cell sensors when supporting the weight of the tubes within the tube support 438, with or without liquid.

[0166] Furthermore, in one embodiment, the tare subsystem 470a and the tare subsystem 470b work together to help the system achieve a relatively low final volume. In one embodiment, the tare subsystem 470b is configured for a high-capacity container with low precision, while the tare subsystem 470a is configured for a low-capacity container with high precision. In this way, for example, the volume transferred from a container corresponding to the tare subsystem 470b can be processed through a chip and placed in a container corresponding to the (high-precision) tare subsystem 470a.

[0167] Furthermore, in another embodiment, the base unit body 420 may be made of aluminum, plastic, and / or stainless steel. However, in other embodiments, the base unit body 420 may be of any material and / or any dimensions not inconsistent with the purposes of this disclosure.

[0168] Figure 5 is a side elevation view showing an example of a base unit without a base unit housing according to the present disclosure. In one embodiment, as shown in Figure 5, the base unit 500 includes a base unit body 520, a first tube holding assembly 532, a second tube holding assembly 534, a third tube holding assembly (not shown), and tare subsystems 570a and 570b. The first tube holding assembly 532, the second tube holding assembly 534, and the third tube holding assembly each have a tube adapter 537 and / or a tube support 538. The tare subsystems 570a and 570b each have a load cell sensor (not shown), one or more linear rails 572, and / or one or more carriages 574. In this way, the base unit 500 can be paused during a protocol run and the linear rails 572 can be lowered. The descent is performed such that the carriage 574 no longer touches the load cell sensor to the tube support 538, and the load cell sensor no longer supports the weight of the tube, regardless of whether there is liquid in the tube support 538, thereby enabling tare of the load cell during execution.

[0169] Furthermore, in one embodiment, the ability to perform tare during a protocol run (e.g., tare the load cell mid-run) means that the effects of load cell drift and creep are reduced. This improves the accuracy of the final volume from the system according to this disclosure. Moreover, this compensates for differences between tubes supplied from the same manufacturer, and this ensures that the integrated tip transfer tube always reaches the bottom of the conical tube or any required depth from the bottom of the tube.

[0170] Furthermore, in one embodiment, when implementing a low-volume concentration protocol, the system can more reliably concentrate the sample to a volume of less than 500 microliters by using the same load cell sensor that was used to determine the initial sample volume for the protocol.

[0171] Figure 6 is a perspective view showing an example of a millifluidic / microfluidic TFF tip with an integrated membrane assembly and fluid handling elements according to the present disclosure. In one embodiment, as shown in Figure 6, the tip 6000 includes a closed fluid channel 6100, a membrane assembly 6300, an integrated transfer tube 6500, and a valve insert 6700. The membrane assembly 6300 has a low-volume channel 6310, a standard-volume channel 6330, and a corresponding concentrating or filtering membrane (not shown, best shown in Figure 7). The valve insert 6700 has a first diaphragm / valve complex 6710 and a second diaphragm / valve complex 6730 interposed along the closed fluid channel 6100. In one embodiment, the positions of the low-volume channel 6310 and the standard-volume channel 6330 may be swapped. However, the position of the low-volume channel 6310 relative to the standard-volume channel 6330 as shown in Figure 6 allows for the lowest possible hold-up volume for the low-volume channel 6310.

[0172] In one embodiment, as shown in Figure 6, the chip 6000 is made of multiple layers of polystyrene, acrylic, or polypropylene, in particular of four rectangular acrylic plastic layers laminated to be approximately 74.0 mm in length and 70 mm in width. However, in other embodiments, these layers may be of any material and / or of any dimensions not inconsistent with the purposes of this disclosure.

[0173] In one embodiment, as shown in Figure 6, the membrane assembly 6300 has a “dual path” in the form of a low-volume path 6310 and a standard-volume path 6330. These are in valve-controlled fluid communication with an integrated transfer tube, one or more closed fluid passages 6100, and other fluid handling elements of the tip 6000. By using the dual path of the tip 6000 individually or in combination, a minimum hold-up volume within the tip 6000 (e.g., as low as approximately 250.0 microliters) can be achieved, and a desired optimized concentration or filtration rate can be achieved. In one embodiment, the low-volume path enables sample concentration up to an industry-leading final output / sample volume of approximately 250.0 microliters.

[0174] Furthermore, in one embodiment, the low-capacity path 6310 has a flow path radius of about 0.4 millimeters. In one embodiment, the standard-capacity path 6330 has a flow path radius of about 0.4 millimeters, but the path is squared off on one side, and the center point of the radius on the curved side opposite the squared side is offset by about 0.1 millimeters. However, in other embodiments, the low-capacity path 6310 and the standard-capacity path 6330 can have any dimensions that are not inconsistent with the purposes of this disclosure.

[0175] Furthermore, in one embodiment, as the initial sample flows through the dual pathway of the membrane assembly 6300, small non-protein molecules, for example, may be filtered out from the tip 6000 along with the permeate and sent to the waste tube of the washing subsystem.

[0176] Returning to Figure 6, in one embodiment, the integrated transfer tube 6500 of the tip 6000 includes a first transfer tube pair 6510, a second transfer tube pair 6530, and a single transfer tube 6550. In one embodiment, when the tip 6000 is engaged with the base unit, the first transfer tube pair 6510 corresponds to a first tube holding assembly (e.g., the first tube holding assembly 232 in Figure 2). The second transfer tube pair 6530 corresponds to a second tube holding assembly (e.g., the second tube holding assembly 234 in Figure 2). The single transfer tube 6550 corresponds to a third tube holding assembly (e.g., the third tube holding assembly 236 in Figure 2).

[0177] In one embodiment, as shown in Figure 6, the integrated transfer tube 6500 is made of 316 stainless steel and fixed to the laminated acrylic plate of the tip 6000 using adhesive. Furthermore, the first transfer tube pair 6510 and the single transfer tube 6550 are defined by an 18RW gauge, while the second transfer tube pair 6530 is defined by a 19RW gauge. However, in other embodiments, the integrated transfer tube 6500 may be of any material and / or any dimensions not inconsistent with the purposes of this disclosure.

[0178] Figure 7 is a front elevation view showing an example of a millifluidic / microfluidic TFF tip with an integrated, individually controllable diaphragm pump and valve according to the present disclosure. In one embodiment, as shown in Figure 7, the tip 7000 includes a closed fluid passage 7100, a low-capacity passage 7310 having a corresponding first membrane 7320, a standard-capacity passage 7330 having a corresponding second membrane 7340, a first transfer tube pair 7510 (T1), a second transfer tube pair 7530 (T2), a single transfer tube 7550 (T3), a valve insert 7700, and an elastomer valve 7780. The valve insert 7700 has a first diaphragm / valve complex 7710 and a second diaphragm / valve complex 7730 interposed along the closed fluid passage 7100. The elastomer valve 7780 is configured as a purge valve (for blowing air through the air passage inlet of the tip to the purge valve outlet; best shown in Figure 3).

[0179] More specifically, in one embodiment, as shown in Figure 7, the valve insert 7700 includes a first diaphragm / valve complex 7710. The first diaphragm / valve complex 7710 has a diaphragm pump 7712 (D2), a first elastomer valve 7714 configured as a T1 input valve, a second elastomer valve 7716 configured as a T2 input valve, and a third elastomer valve 7718 configured as a D2 output valve.

[0180] Furthermore, in one embodiment, as shown in Figure 7, the valve insert 7700 includes a second diaphragm / valve complex 7730. The second diaphragm / valve complex 7730 comprises a diaphragm pump 7732 (D2), a first elastomer valve 7734 configured as a T1 input valve, a second elastomer valve 7736 configured as a T2 input valve, a third elastomer valve 7738 configured as a D2 output valve, and a fourth elastomer valve 7739 configured as a T3 input valve.

[0181] Furthermore, in one embodiment, as shown in Figure 7, the valve insert 7700 of the tip 7000 also includes an elastomer valve 7750 configured as a T1 return valve, an elastomer valve 7755 configured as a T2 return valve, an elastomer valve 7760 configured as an input valve to the low-capacity path 7310 and the first membrane 7320, an elastomer valve 7765 configured as an input valve to the standard-capacity path 7330 having a corresponding second membrane 7340, an elastomer valve 7770 configured as an output valve from the low-capacity path 7310 and the first membrane 7320 (M1), and an elastomer valve 7775 configured as an output valve from the standard-capacity path 7330 having a corresponding second membrane 7340 (M2).

[0182] Furthermore, in one embodiment, four input valves independently connect T1 and T2 to D1 and D2, thereby enabling selective suction or discharge of liquid to or from either T1 or T2. Also in one embodiment, D1 and D2 are configured to independently suction or discharge liquid synchronously or asynchronously, periodic or non-periodicly. Also in one embodiment, the output valves of D1 and D2 are configured to perform independently controlled actuation / stroke and to act synchronously with D1 and / or D2, opening a portion of the closed fluid passage toward either M1 and / or M2 to control the filtration pressure. Also in one embodiment, four membrane valves are configured to selectively open and close portions of the closed fluid passage leading to M1 or M2 (e.g., any fluid path toward M1, M2, or both can be opened). Also in one embodiment, two return valves are configured to return to either T1 or T2 and are independently pressure-controlled to restrict flow in order to control the filtration pressure. In one embodiment, the T3 input valve is configured to open the path to T3, thereby allowing additional sample or buffer to be transferred to either T1 or T2. In another embodiment, the purge valve is configured to blow air into one or more closed fluid channels of the tip at the end of the sequence to prevent excess liquid from dripping after the sample processing is complete.

[0183] Furthermore, in one embodiment, the T3 input valve is configured to drive a flow across both membrane paths by D2, but in the reverse direction (e.g., drawn in from T3 across M2 and M1). Also in one embodiment, the input and output valves of M1 and M2, as well as the D2 output valve, remain open at all times while the reverse protocol is running. Also in one embodiment, depending on the destination tube, either the D2 output valve, the T3 input valve, and either the T1 or T2 return valve open or close to draw in from T3 and then push out to either T1 or T2. Also in one embodiment, the T3 input valve may be connected to D2 via a new channel instead of crossing the membrane. Also in one embodiment, one or more diaphragm pumps may drive the reverse flow across the membrane.

[0184] Furthermore, in one embodiment, as shown in Figures 2, 3, 6, and 7, the tip 7000 is configured to automatically, efficiently, and effectively rinse, clean, and / or sterilize the fluid handling elements of the tip, particularly the closed fluid channel 7100, the low-volume channel 7310, the standard-volume channel 7330, and the integrated transfer tubes for T1, T2, and T3. In one embodiment, a base unit as shown in Figures 2-5 is configured to clean, rinse, and / or sterilize the fluid handling elements of the tip 7000, or a separate assembly or subsystem of a broader system is configured to do the same. In any case, in one embodiment, the base unit or cleaning subsystem is configured to autonomously meter one or more cleaning fluids and / or equilibration buffers (and air, as described herein for example with respect to Figure 3) (at any point in the protocol, run, method, or procedure), actuate diaphragm pumps and valves to appropriately flow or drive the fluid or liquid (or air), and receive, recover, and / or purge the cleaning output from the tip 7000. Thus, in one embodiment, the base unit or cleaning subsystem is configured to deliver the buffer or any other liquid or fluid with a peristaltic pump. Furthermore, in one embodiment, the base unit or cleaning subsystem is configured to first flow one or more cleaning fluids into the fluid handling element of the tip 7000, then purge the fluid handling element of the tip, then rinse with a neutral buffer, then flow air into the fluid handling element of the tip, and then capture the cleaning output (either solid, liquid, and / or gas) from the tip 7000.

[0185] As background, with conventional TFF chips or cassettes, users must set up and run a protocol using sodium hydroxide, then disassemble the setup and chip, purge all lines, and repeat this process with a neutralizing buffer. Furthermore, disassembly and purging of all lines must be performed again before introducing the sample.

[0186] Furthermore, in one embodiment, the base unit or cleaning subsystem uses an external reservoir of cleaning solution and buffer / water, as well as a peristaltic pump, to sterilize and filter one or more tips. Two peristaltic pumps have tubing extending to two interchangeable quick-connect needles that are in fluid communication with the tips on the base unit or cleaning subsystem, and can, for example, flush one or more tips with one or more cleaning solutions, sterilize them, or remove preservatives. The cleaning solution is then pumped out of the tips to a drain or waste line, and one or more tips can be flushed with buffer or water to remove one or more cleaning solutions. In one embodiment, the base unit or cleaning subsystem cleans one or more tips using one or more preferred cleaning solutions and one or more rinse solutions (e.g., buffer, water) pre-selected / programmed by the user.

[0187] Figure 8 is an enlarged cutaway perspective view showing an example of a valve insert for a millifluid / microfluid TFF tip at a limit point according to the present disclosure. In one embodiment, as shown in Figure 8, the tip 8000 includes a closed fluid passage 8100 and a valve insert 8700. The valve insert 8700 has an elastomer valve 8750 configured as a T1 return valve and as a first limit point of the fluid handling element of the tip 8000, and an elastomer valve 8755 configured as a T2 return valve and as a second limit point of the fluid handling element of the tip 8000. Furthermore, in one embodiment, by controlling / closing / limiting these limit points, the tip 8000 increases the back pressure on the flow over M1 and / or M2, thereby promoting faster concentration, and by opening, achieves the opposite effect.

[0188] Furthermore, in one embodiment, the limiting point may have a shape different from those illustrated in Figures 7 and 8, such as a valve shape (e.g., oval). However, the following limitations should be noted. Flutter: When the valve is highly restricted, it essentially flutters between open and fully closed, producing an audible squeaking sound and causing pressure fluctuations in the upstream flow. While functional, this is not ideal. Dynamic Range: By redesigning the shape as shown in Figures 7 and 8, the Tip 8000 eliminates flutter, allowing the valve to operate better at higher pressures and providing a wider dynamic range in trans-membrane pressure operations.

[0189] Furthermore, in one embodiment, the limit point maintains constant pressure on the upstream side and the downstream side of the closed fluid passage of the tip. Also, in one embodiment, the molded limit point, as shown in Figures 7 and 8, has multiple smaller holes and a circular / domed valve design to improve linearity and fluttering (e.g., pressure spikes).

[0190] Figure 9 is a perspective view showing an example of a valve insert for a millifluidic / microfluidic TFF tip according to the present disclosure. In one embodiment, as shown in Figure 9, the valve insert 9700 of the tip 9000 includes a first diaphragm / valve complex 9710, a second diaphragm / valve complex 9730, a T3 input valve 9739, a T1 return valve 9750, a T2 return valve 9755, an input valve 9765 to a low-capacity path of a membrane assembly (not shown), an input valve 9765 to a standard-capacity path of a membrane assembly (not shown), an output valve 9770 from the low-capacity path, and an output valve 9775 from the standard-capacity path. The first diaphragm / valve complex 9710 has a diaphragm pump 9712, a T1 input valve 9714, a T2 input valve 9716, and a D2 output valve 9718. The second diaphragm / valve complex 9730 includes a diaphragm pump 9732 (D2), a T1 input valve 9734, a T2 input valve 9736, and a D2 output valve 9738.

[0191] Furthermore, in one embodiment, the valve insert 9700 is made of molded silicone and defines a protruding oval rim (for the valve) and a spherical / domed recess (for the diaphragm pump), and has dimensions of approximately 0.4 mm × approximately 0.6 mm. However, in other embodiments, the valve insert 9700 may be made of any material and / or have any dimensions that are not inconsistent with the purposes of this disclosure.

[0192] Furthermore, in one embodiment, the valve insert 9700 is configured to elastically deform in response to airflow or pneumatic pressure in order to effectively open and close / stroke a diaphragm pump and valve in a fully assembled tip, for example. In one embodiment, the valve insert is configured to elastically deform by a mechanical pusher, without requiring pneumatic operation.

[0193] Figure 10A is a first perspective view showing an example of an automation subsystem relating to this disclosure, including a mobile gantry with a robotic gripper.

[0194] Figure 10B is a second perspective view showing an example of the automation subsystem in Figure 10A.

[0195] Figure 11 is a perspective view showing an example of an automation subsystem relating to the present disclosure, which includes a gantry configured to move vertically and has a robotic gripper that operates in conjunction with a sliding pivot point on the gantry.

[0196] Figure 12 is a perspective view showing an example of a gantry and one or more tube racks relating to this disclosure.

[0197] Figure 13 is a perspective view showing an example of one or more automated tube racks and / or chips / one or more cassette racks relating to this disclosure.

[0198] Figure 14 is a perspective view showing an example of one or more automated tube racks and / or chips / one or more cassette racks relating to this disclosure.

[0199] Figure 15 is a perspective view showing an example of one or more automated tube racks and / or chips / one or more cassette racks relating to this disclosure. IV. Embodiments

[0200] Article 1. This is a tangential flow filtration (TFF) tip, (a) Chip substrate assembly defining a closed fluid channel, (b) A membrane assembly comprising the following: (i) Concentration membrane or filtration membrane, (ii) Low volume pathways, and (iii) Standard capacity path, (c) Integrated transfer tube, and (d) comprising one or more diaphragm / valve complexes located on a valve insert and interposed along the closed fluid passage, The closed fluid channel is in fluid communication with the low-volume path and the standard-volume path under valve control, in a tangential flow filtration (TFF) tip.

[0201] Article 2. The TFF tip according to Clause 1, wherein each of the one or more diaphragm / valve complexes comprises one diaphragm pump and three individually controllable elastomer valves.

[0202] Article 3. The TFF tip according to Clause 2, wherein one of the three individually controllable elastomer valves is the first source input valve.

[0203] Article 4. One of the three individually controllable elastomer valves is the second source input valve, as described in Clause 2 of the TFF tip.

[0204] Article 5. One of the three individually controllable elastomer valves is a diaphragm output valve, as described in Clause 2 of the TFF tip.

[0205] Article 6. The valve insert further comprises a third source input valve, as described in Clause 1, for the TFF tip.

[0206] Article 7. The valve insert further comprises a first supply source return valve, The TFF tip according to Clause 1, wherein the first source return valve, when closed, acts as a limiting point that promotes faster concentration due to the back pressure on the flow over the concentration or filtration membrane.

[0207] Article 8. The valve insert further comprises a second supply source return valve, The TFF tip according to Clause 1, wherein the second source return valve, when closed, acts as a limiting point that promotes faster concentration due to the back pressure on the flow over the concentrating or filtering membrane.

[0208] Article 9. The valve insert further comprises a low-capacity path input valve and a low-capacity path return valve, as described in Clause 1.

[0209] Article 10. The valve insert further comprises a standard capacity path input valve and a standard path return valve, as described in Clause 1.

[0210] Article 11. A TFF tip as described in Clause 1, further equipped with a purge valve.

[0211] Article 12. A high-throughput automated multi-sample processing system, (a) Base unit, (b) A tangential flow filtration (TFF) chip configured to operate with the base unit, the TFF chip comprising: (i) Chip substrate assembly defining a closed fluid channel, (ii) A membrane assembly comprising (i) a concentrating membrane or a filtration membrane, (ii) a low-volume pathway, and (iii) a standard-volume pathway, (iii) Integrated transfer tube, and (iv) One or more diaphragm / valve complexes interposed along the closed fluid passage, (c) an automation subsystem for engaging with the TFF chip, and (d) A housing that defines the envelope (outer shell) of the system.

[0212] Article 13. The system as described in Clause 12, further comprising a chip rack within an automated bay for holding one or more chips.

[0213] Article 14. The system described in Clause 12 further includes a tube rack within the automated bay.

[0214] Article 15. The system described in Clause 12 further comprises a cleaning subsystem within the automated bay.

[0215] Article 16. The automation subsystem comprises a gantry and a robotic gripper, as described in Clause 12.

[0216] Article 17. The system according to Clause 16, wherein the automation subsystem is configured to move above the envelope of the housing.

[0217] Article 18. The gantry is a movable gantry, as described in Clause 17.

[0218] Article 19. The movable gantry is configured to move left and right within the envelope of the housing, or to move up and down within the envelope of the housing. The system according to Clause 18, wherein the movable gantry is configured such that the pivot point of the robotic gripper moves up and down or left and right within the envelope of the housing.

[0219] Article 20. The base unit comprises a first tare subsystem and a second tare subsystem, The system according to Clause 12, wherein the second tare subsystem is a subsystem with higher accuracy compared to the first subsystem.

[0220] It should be emphasized that the embodiments described above are merely practical examples presented to clearly illustrate the principles of this disclosure. Many variations and modifications can be made to one or more of the embodiments described above without substantially departing from the spirit and principles of this disclosure. All such modifications and variations are intended to be within the scope of this disclosure and protected by the following claims.

Claims

1. A tangential flow filtration (TFF) tip, a. Chip substrate assembly, b. Membrane assembly, c. Integrated transfer tube, and d. comprising one or more diaphragm / valve complexes, The aforementioned chip substrate assembly defines a closed fluid channel, The aforementioned film assembly is i. Concentration membrane or filtration membrane, ii. Low-capacity pathways, and iii. Equipped with a standard capacity path, The one or more diaphragm / valve complexes are located on a valve insert and interposed along the closed fluid passage, The closed fluid passage is in fluid communication with the low-capacity path and the standard-capacity path under valve control. TFF chip.

2. A TFF chip according to claim 1, Each of the one or more diaphragm / valve complexes comprises one diaphragm pump and three individually controllable elastomer valves. TFF chip.

3. A TFF chip according to claim 2, One of the three individually controllable elastomer valves is the first supply source input valve. TFF chip.

4. A TFF chip according to claim 2, One of the three individually controllable elastomer valves is the second supply source input valve. TFF chip.

5. A TFF chip according to claim 2, One of the three individually controllable elastomer valves is a diaphragm output valve. TFF chip.

6. A TFF chip according to claim 1, The valve insert further comprises a third supply source input valve. TFF chip.

7. A TFF chip according to claim 1, The valve insert further comprises a first supply source return valve, When the first supply source return valve is closed, the back pressure on the flow over the concentrating or filtering membrane acts as a limiting point that promotes faster concentration. TFF chip.

8. A TFF chip according to claim 1, The valve insert further comprises a second supply source return valve, When the second supply source return valve is closed, the back pressure on the flow over the concentrating or filtering membrane acts as a limiting point that promotes faster concentration. TFF chip.

9. A TFF chip according to claim 1, The valve insert further comprises a low-capacity path input valve and a low-capacity path return valve. TFF chip.

10. A TFF chip according to claim 1, The valve insert further comprises a standard capacity path input valve and a standard path return valve. TFF chip.

11. A TFF chip according to claim 1, It also has a purge valve. TFF chip.

12. A high-throughput automated multi-sample processing system, a. Base unit, b. Tangential flow filtration (TFF) tip, c. Automation subsystem, and d. Housing, equipped with The TFF chip is configured to operate together with the base unit. The aforementioned TFF chip is i. Chip substrate assembly, ii. Membrane assembly, iii. Integrated transfer tube, and, iv. Equipped with one or more diaphragm / valve complexes, The aforementioned chip substrate assembly defines a closed fluid channel, The aforementioned film assembly is I. Concentration membrane or filtration membrane, II. Low-volume pathways, and III. Equipped with a standard capacity path, The one or more diaphragm / valve complexes are interposed along the closed fluid passage, The automation subsystem is for engaging with the TFF chip, The housing defines the envelope of the system. system.

13. The system according to claim 12, Further equipped with chip racks in the automated bay for holding one or more chips, system.

14. The system according to claim 12, With additional tube racks in the automated bay, system.

15. The system according to claim 12, Further equipped with a cleaning subsystem in the automated bay, system.

16. The system according to claim 12, The automation subsystem comprises a gantry and a robotic gripper. system.

17. The system according to claim 16, The automation subsystem is configured to move above the envelope of the housing. system.

18. The system according to claim 17, The aforementioned gantry is a movable gantry. system.

19. The system according to claim 18, The movable gantry is configured to move left and right within the envelope of the housing, or to move up and down within the envelope of the housing. The movable gantry is configured such that the pivot point for the robot gripper moves up and down or left and right within the envelope of the housing. system.

20. The system according to claim 12, The base unit comprises a first tare subsystem and a second tare subsystem, The second tare subsystem is a more accurate tare subsystem compared to the first tare subsystem. system.

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