Patent Application
20250213757
The present disclosure is directed to the processing of biological fluid using a disposable fluid circuit and a reusable processing device to generate a product and a method of processing a biological fluid. More particularly, the present disclosure is directed to the processing of biological fluid with sequential incubation stages, resulting in sequential incubation products.
The processing of biological fluid such as blood or blood components typically involves using a reusable processing device (“hardware”) and a disposable fluid circuit adapted for mounting or other association with the reusable device. The fluid circuit typically includes (plastic) bags and associated tubing that defines a flow path through the circuit. The disposable fluid circuit may also include one or more separation devices where the biological fluid/cells can be separated into two or more components, washed or otherwise processed. Separation devices may separate the biological fluid based on centrifugal separation and/or, as described below, membrane separation.
Occasionally, it is desired to include an incubation stage or stages when processing a biological fluid. Current systems require separation stages between successive incubation stages or require a manual step separate from the system from an additional incubation step. However, there are certain products that require sequential incubation. The system and method described herein provides for automated sequential incubation stages in an automated fluid processing system and method.
In one aspect, a fluid processing system includes a source container, a fluid circuit connectable to the source container, at least one pump, at least one clamp, and a controller. The fluid circuit includes a separator, an in-process container, and tubing connecting the components of the fluid circuit. The controller is coupled to the at least one pump, separator, and the at least one clamp. The controller is configured to actuate the at least one pump, separator, and the at least one clamp to sequentially execute a first incubation stage and a second incubation stage within an incubation loop that includes the separator, the in-process container, and tubing which connects the in-process container and the separator.
According to another aspect, a fluid processing method includes conveying a source fluid to an in-process container, executing a first incubation stage, and executing a second incubation stage. The first incubation stage includes adding a first fluid reagent to the source fluid in the in-process container to create a first fluid product and mixing the first fluid product to generate a first incubation fluid product. The second incubation stage includes adding a second fluid reagent to the in-process container and the first incubation fluid product to create a second fluid product and mixing the second fluid product to generate a second incubation fluid product. The first and second incubation stages are executed sequentially without any intervening steps.
The disclosure will be more fully understood from the following description taken in conjunction with the accompanying drawings. Some of the figures may have been simplified by the omission of selected elements for the purpose of more clearly showing other elements. Such omissions of elements in some figures are not necessarily indicative of the presence or absence of particular elements in any of the exemplary embodiments, except as may be explicitly delineated in the corresponding written description. None of the drawings is necessarily to scale.
As illustrated in
As explained in detail below, the system 10 includes a disposable processing fluid circuit 100 (see also
Thus, the fluid processing systems disclosed herein typically include a reusable separation device and one or more disposable processing circuits adapted for association with the reusable device, which device and circuit(s) are operated by a controller. The reusable separation device may be any device that can provide for the automated processing of biological fluid. “Biological fluid” includes without limitation blood and blood components, and blood cells, such as red cells, white cells and platelets. By “automated,” it is meant that the device can be programmed to carry out the processing steps of a biological fluid processing method without substantial operator involvement. Of course, even in the automated system of the present disclosure, it will be understood that operator activity may be involved, including the loading of the disposable fluid circuits and entering processing parameters. Additional manual steps may be required as well. However, the reusable device can process biological fluid through the disposable circuit(s) described below without substantial operator intervention.
The illustrated processing device is typically capable of effecting the separation of a biological fluid that includes biological cells into two or more components or fractions. Thus, the device may generate conditions that allow for the separation of a biological fluid into selected components or fractions. The device may also be used to perform multiple incubations.
One preferred device for separating biological fluid into its constituent components or fractions uses a spinning porous membrane. An example of such device is the Autopheresis CR sold by Fenwal, Inc. of Lake Zurich, Illinois, which is an affiliate of Fresenius Kabi AG of Bad Homburg, Germany. A detailed description of a spinning membrane may be found in U.S. Pat. No. 5,194,145 to Schoendorfer, which is incorporated by reference herein in its entirety, and in International (PCT) Application No. PCT/US2012/028492, filed Mar. 9, 2012, the contents of which are also incorporated herein in their entirety. In addition, systems and methods that utilize a spinning porous membrane are also disclosed in U.S. Provisional Patent Application No. 61/537,856, filed on Sep. 22, 2011, and International (PCT) Application No. PCT/US2012/028522, filed Mar. 9, 2012, the contents of each are incorporated herein by reference. A further example of a cell or component processing system that utilizes a spinning membrane is the Lovo Cell Processing System which is described in U.S. Pat. Nos. 11,666,687; 10,781,001; 11,827,398; 11,191,879; 10,654,000; and 10,449,263, the contents of each are incorporated herein by reference.
The references identified above describe a membrane-covered spinner having an interior collection system disposed within a stationary shell. While a detailed discussion of the separation device is beyond the scope of this application, the spinning membrane separation device is shown in
Turning now to
As will be seen in the Figures and described in detail below, the disposable fluid processing circuits include tubing that defines flow paths throughout the circuits, as well as access sites for sterile or other connection to containers of processing solutions, such as wash solutions, treating agents, or sources of biological fluid. As shown in
As will be apparent from the disclosure herein, source containers may be attached in sterile fashion to the circuit 100. Source containers 102 for connection to one disposable circuit may be the product containers 150 of another circuit used in an earlier step of the overall method of processing. Alternatively, the contents of a product container 150 may be further processed or separated and then transferred in sterile fashion to the source container 102 of a later-in-series fluid circuit.
The biological fluid to be treated is typically provided in a source container 102, shown in
As further shown in
In accordance with the fluid circuit of
Tubing segment 136 defines a flow path connected at one end to branched-connector 126 and to an inlet port 20 of the separator 101 or in-process container 122. Preferably, in accordance with the present disclosure, separation device 101 is a spinning membrane separator of the type described in U.S. Pat. Nos. 5,194,145 and 5,053,121, which are incorporated by reference, U.S. Provisional Patent Application Ser. No. 61/451,903 and PCT/US2012/028522, also previously incorporated herein by reference.
As shown in
Separation device 101 preferably includes a second outlet 48 that is connected to tubing segment 142 for directing the desired biological cell/fluid product to the in-process container(s) 122 or the product container 150. To permit this, the other end of tubing segment 142 is connected to branched-connector 144, which branches into and defines a flow path to one or more in-process containers 122 and a flow path to a “final” product container 150. The product container 150 may also include a sampling assembly (not shown).
Shown in greater detail in
Device 200 may be of compact size suitable for placement on a tabletop of a lab bench and adapted for easy transport. Alternatively, device 200 may be supported by a pedestal that can be wheeled to its desired location. In any event, device 200 includes a plurality of peristaltic pumps such as pumps 202, 204 and 206 on front panel 201. Pump segments of the disposable fluid circuit (described above) are selectively associated with peristaltic pumps 202, 204, and 206. The peristaltic pumps articulate with the fluid set of
Device 200 may also include several sensors to measure various conditions. The output of the sensors is utilized by device 200 to operate one or more wash or processing cycles. One or more pressure transducer sensor(s) 226 may be provided on device 200 and may be associated with a disposable set 100 at certain points to monitor the pressure during a procedure. Pressure transducer 226 may be integrated into an in-line pressure monitoring site (at, for example, tubing segment 136), to monitor pressure inside separator 101. Air detector sensor 238 may also be associated with the disposable set 100, as necessary. Air detector 238 is optional and may be provided to detect the location of fluid/air interfaces.
Device 200 may also include weight scales 240, 242, 244, and 246 from which the final product container 150, waste container 140, the source container 102 and the in-process container 122, respectively, may depend and be weighed. The weights of the bags are monitored by weight sensors and recorded during a washing or other procedure. From measurements of the weight sensors, the device determines whether each container is empty, partially full, or full and controls the components of device 200, such as the peristaltic pumps 202, 204 and 206 and clamps 210, 212, 214, 216, 218, 220 and 222.
Device 200 includes at least one drive unit or “spinner” 248, which causes the indirect driving of the spinning membrane separator 101. Spinner 248 may consist of a drive motor connected and operated by device 200, coupled to turn an annular magnetic drive member including at least a pair of permanent magnets. As the annular drive member is rotated, magnetic attraction between corresponding magnets within the housing of the spinning membrane separator causes the spinner within the housing of the spinning membrane separator to rotate.
Turning to
Device 101 includes a generally cylindrical housing 12, mounted concentrically about a longitudinal vertical central axis. An internal member 14 is mounted concentric with the central axis 11. Housing 12 and internal member 14 are relatively rotatable. In the preferred embodiment, as illustrated, housing 12 is stationary and internal member 14 is a rotating spinner that is rotatable concentrically within cylindrical housing 12, as shown by the thick arrow in
The shear gap also may vary along the axial direction, for example preferably an increasing gap width in the direction. Such a gap width may range from about 0.02 to about 0.075 inches (0.05-0.19 cm). The gap width could be varied by varying the outer diameter of the rotor and/or the inner diameter of the facing housing surface. The gap width could change linearly or stepwise or in some other manner as may be desired. In any event, the width dimension of the gap is preferably selected so that at the desired relative rotational speed, Taylor-Couette flow, such as Taylor vortices, are created in the gap.
With reference to
Cylindrical housing 12 is completed by a bottom end housing terminating in an outlet orifice 46 concentric with the central axis. The outlet orifice 46 may also include a cap, clamp, or other closure mechanism, which can be utilized to close the orifice 46 during an incubation step in which nothing is exiting the separator at this outlet. In one embodiment, the instrument clamp 222 on the tubing line 138 is used to stop the fluid flow.
In the illustrated embodiment, the surface of the rotary spinner 14 is at least partially, and is preferably substantially or entirely, covered by a cylindrical porous membrane 62. The membrane 62 may have a nominal pore size between 0.8 and 10 microns (μm), for example. Membranes may be fibrous mesh membranes, cast membranes, track-etched membranes or other types of membranes that will be known to those of skill in the art. For example, in one embodiment, the membrane may have a polyester mesh (substrate) with nylon particles solidified thereon, thereby creating a tortuous path through which only certain sized components will pass. In an embodiment, the nylon membrane may have a pore size of approximately 0.8 μm and a thickness of approximately 150 μm or greater. Membranes of this type will typically retain all cellular components (e.g., red blood cells, white blood cells) and certain formed blood components, e.g., platelets. In another embodiment, the membrane may be made of a thin (approximately 10 μm thick) sheet of unsupported polycarbonate, for example, with a pore size of approximately 4.0 μm. In this embodiment, pores (holes) may be cylindrical and larger than those described above. The pores may be sized to allow small formed components (e.g., platelets, microparticles, etc.) to pass, while the desired cells (e.g., white blood cells and larger red blood cells) are collected.
Having thus described the system, including disposable circuit 100 and reusable device 200, reference is made to
As is also illustrated in
The at least one input 302 may include a number of different devices according to the embodiments described herein. For example, the input 302 could include a keyboard or keypad by which a user may provide information and/or instructions to the controller 300. Alternatively, the input 302 may be a touch screen, such as may be used in conjunction with a video display 308 that is disposed on the front panel 201 of the device 200, the video display 308 also being coupled to the controller 300. The input could also include a reader or scanner, such as a barcode reader or scanner or an RFID reader. The assembly of the input/touch screen 302 and video display 308 may be one of the afore-mentioned structures to which the controller 300 is coupled from which the controller 300 receives information and to which the controller 300 provides commands. According to still other embodiments, the input 302 may be in the form of computer equipment that permits the fluid processing system including the controller 300 to communicate (whether via wires, cables, etc. or wirelessly) with other fluid processing systems over a local network, or with other fluid processing systems or other computer equipment (e.g., a server) over local networks, wide area networks, or the Internet. According to such an embodiment, the input may include an internal transmitter/receiver device.
Having discussed the structure of embodiments of the fluid processing system disclosed herein, the operation of the fluid processing system is now discussed. The fluid processing method steps of the current disclosure may be performed alone or within a larger fluid processing method.
In general, the operator may first activate (e.g., switch on) the device 200, at which point the device 200 conducts self-calibration checks, including the checking of the peristaltic pumps 202, 204, 206, clamps 210, 212, 214, 216, 218, 220, 222, and sensors 226, 238. Device 200 may then prompt the user to enter or modify process parameters using the input 302, including by way of example and not by way of limitation: number of incubations within each cycle, incubation volume, incubation wash media, incubation duration, incubation flow rate, incubation spinner revolution rate, pre- and post-incubation pauses, and pre- and post-incubation pause text. The number of incubations can vary from 0 (which is disabled), 1 (single) or 2 (double/sequential). The incubation volume can vary from 5-800 mL, and can include the ability to target the minimum achievable incubation volume based on all procedure inputs and estimations. The specific incubation wash media can be selected, such as, solution 1 or solution 2, which is the user provided solution used by the system to pump the cell suspension to the incubation volume, these solutions being attached at 123. The incubation can be varied from 1-99 minutes. The incubation flow rate, or flow rate through the incubation loop, can vary from 0-200 mL/min, wherein 0 indicates no flow and off-line agitation via a rocker tray. The incubation spinner revolution rate can be between 0-4500 RPM (0 if not being used to circulate or mix the incubation solution). There can be pre- or post-incubation pauses and during these pauses the text on the screen may indicate instructions for the user to follow. The device 200 may then prompt the operator to mount the disposable set 100, after which device 200 automatically checks to determine whether the disposable set 100 is properly installed. Once the set 100 is properly installed, the controller 300 prompts the operator to connect the source of biological fluid (e.g., 102 of
Once the operator confirms that the solutions are connected, the controller 300 primes the disposable set 100. In the embodiment discussed above, the set 100 may be primed with saline, although other biocompatible aqueous solutions may also be used. The controller then commences the biological fluid processing. The controller may direct the source fluid directly to the in-process container or perform a separation before. If a separation is done, the controller 300 then initiates the transferring from source container (e.g., 102 of
If an initial separation is not desired, the biological source material may be transferred to the in-process container 122.
Once biological cells are collected in the in-process bag 122, either from a source container or as a separation product, a reagent or solution may be added to the biological cells in the in-process bag to initiate a first incubation stage. This can be done at port 123 of the in-process bag, such as by syringe or other transfer device. The reagent or solution can be any known reagent or solution depending on the incubation process desired. The reagents can be reagents containing antibody complexes that allow for specific labeling of desired cells in suspension. For example, the reagent may include antibody complexes recognizing CD4+ cells and magnetic particles. Once the target cells (CD4+) are labeled they can be removed from the remaining cell suspension with a magnet. The combination of the reagent or solution and the biological cells of the source fluid produce a first fluid product.
This first fluid product is then mixed or agitated to generate a first incubation fluid product. There may be a waiting period before this agitation or mixing is started. The waiting period can be 15 min, 30 min, 1 hr, or any amount of time chosen by the user/operator. The waiting period can also be 0 minutes, or the mixing is initiated immediately after the introduction of the reagent or solution. This mixing can be accomplished by circulating the first fluid product from the in-process bag 122 to the separator and eventually back to the in-process bag 122 or can be done by agitating with an external device offline, such as a rocker tray or by manual mixing where the user inverts the bag repeatedly. The mixing step can be done for a set amount of time, such as between 1 and 99 minutes or 10-75 mins, or 25-50 minutes. The set amount of time can be any integer between 1-99. The flow of the liquid through the separator and back to the in-process bag can also be regulated to a certain speed, such as between 0 mL/min and 200 mL/min, the speed can be between 50 mL/min and 150 mL/min or between 75 mL/min and 125 mL/min. The flow rate can be any integer between 0 and 200. Once the first incubation fluid product is adequately mixed, the first incubation stage ends and the first incubation fluid product is in the in-process bag 122.
Immediately after the first incubation stage a second incubation stage is executed. The first incubation fluid product is in the in-process bag 122 of the fluid circuit. A second fluid reagent or solution is added to the first incubation fluid product at the port 123 of the in-process bag 122. This can be done at port 123 of the in-process bag 122, such as by syringe or other transfer device. The reagent or solution can be any known reagent or solution depending on the incubation process desired. The combination of the reagent or solution and the first incubation fluid product of the source fluid produces a second fluid product.
This second fluid product is then mixed or agitated to generate a second incubation fluid product. There may be a waiting period before this agitation or mixing is started. The waiting period can be 15 min, 30 min, 1 hr, or any amount of time chosen by the user/operator. The waiting period can also be 0 minutes, or the mixing is initiated immediately after the introduction of the reagent or solution. This mixing can be accomplished by circulating the first fluid product from the in-process bag 122 to the separator and eventually back to the in-process bag 122 or can be done by agitating with an external device offline, such as a rocker tray. The mixing step can be done for a set amount of time, such as between 1 and 99 minutes or 10-75 mins, or 25-50 minutes. The set amount of time can be any integer between 1-99. The flow of the liquid through the separator and back to the in-process bag can also be regulated to a certain speed, such as between 0 mL/min and 200 mL/min, the speed can be between 50 mL/min and 150 mL/min or between 75 mL/min and 125 ml/min. The flow rate can be any integer between 0 and 200. Once the second incubation fluid product is adequately mixed, the first incubation stage ends and the second incubation fluid product is in the in-process bag 122.
The first and second incubation stages are executed sequentially and without any intervening steps.
After the second incubation stage, a separation stage may be performed, the resulting second incubation fluid product may be washed, or the in-process bag may be removed with the second incubation fluid product.
If a separation stage is executed, the second incubation fluid product is transferred from the in-process bag 122 through the set to the spinning membrane separator 101 via the operation of one or more peristaltic pumps 202, 204 and 206. In a similar fashion, the wash medium is delivered from its container (e.g., 135a, 135b of
Once the processing is completed, the controller prompts the operator to sample, seal and remove the product container 150. The resulting product is a washed, concentrated, and purified cell suspension.
The systems and methods described herein may be effective, for example, in cell enrichment processing routines. These processing routines include a separation stage and at least two incubation stages.
The fluid processing system may support enhanced CD34+ cell enrichment processing routines. Specifically, to support the enrichment of CD34+ cell populations within peripheral blood mononuclear cell (PBMC) apheresis products. The system described above will first execute a separation process to remove platelets which could negatively impact binding an antibody conjugate. The system then incubates the platelet-depleted cell suspension with an Immunoglobulin G (lgG) solution to reduce the incidence of non-specific binding of an antibody conjugate [new functionality]. The system then incubates the platelet-depleted, IgG-incubated cell suspension with an antibody conjugate. The resulting product is a washed, concentrated, and purified cell suspension.
While the foregoing discussion references embodiments in the form of a fluid processing system, other systems may incorporate this technology as well. These systems may share the technical challenges faced by the aforementioned fluid processing system, and incorporation of the technology may provide similar advantages. For example, a separation system, more particularly a filtration system, or even more particularly a microfiltration system, also may include a processor to receive a fluid to be processed and a controller. Further, certain embodiments of such a processor may include a disposable fluid circuit (which circuit may include a membrane used for filtration) and reusable hardware, and the controller may be configured to operate the processor.
Thus, an improved method and system have been disclosed for the sequential incubating and processing of a fluid. The description provided above is intended for illustrative purposes only and is not intended to limit the scope of the invention to any specific method, system, or apparatus, or device described herein except as may be explicitly delineated above.
This application claims the benefit of and priority of U.S. Provisional Patent Application Ser. No. 63/615,143, filed Dec. 27, 2023, the contents of which are incorporated by reference herein.
| Number | Date | Country | |
|---|---|---|---|
| 63615143 | Dec 2023 | US |
