阅读说明：本技术 交替切向流泵送方法 (Alternating tangential flow pumping method ) 是由 鲁道夫·帕弗里克 于 2020-05-21 设计创作，主要内容包括：本公开总体上涉及交替切向流(ATF)灌注泵送方法,包括用于在使用时产生正压和负压的新方法,并且更具体地涉及设备、系统和使用该设备、系统的方法。(The present disclosure relates generally to Alternating Tangential Flow (ATF) perfusion pumping methods, including novel methods for generating positive and negative pressures when in use, and more particularly to devices, systems, and methods of using the same.)

1. An alternating tangential flow pressure device comprising:

a pneumatic cylinder having a proximal end and a distal end and comprising a chamber, the proximal end connected in series to a filter system, wherein the proximal end comprises an opening to the chamber; and

a piston connecting link entering the pneumatic cylinder through the distal end.

2. The apparatus of claim 1, wherein the piston further comprises a linear servo.

3. The apparatus of claim 2, wherein the linear servo further comprises an encoder.

4. The apparatus of claim 3, wherein the position of the linear servo provides information about the action occurring within the filter system.

5. An alternating tangential flow pumping method comprising:

connecting a pneumatic cylinder to a filter ball base, the pneumatic cylinder having a proximal end including a central opening, a distal end including a piston, and a chamber, the filter ball base having a central opening with the opening of the proximal end aligned with the opening of the ball base;

driving the piston with a linear servo; and

moving the piston into and out of the cylinder, creating positive and negative pressure.

6. The method of claim 5, further comprising tracking movement of the piston with an encoder connected to the linear servo.

7. An alternating tangential flow pumping system comprising:

a pneumatic cylinder having a proximal end and a distal end, and comprising a chamber, the proximal end connected in series to a filter system, wherein the proximal end comprises an opening in the center of the proximal end, the opening leading to the chamber;

a piston connecting link entering the pneumatic cylinder through the distal end;

the filter system, wherein the system comprises a sphere comprising a first hemisphere and a second hemisphere, wherein a septum is between the first hemisphere and the second hemisphere, the first hemisphere connected to a distal end of a cylinder, wherein a proximal end of the cylinder is connected to a liquid source, and the cylinder contains a filter.

8. The system of claim 6, wherein the filter system comprises more than one sphere and cylinder.

9. The system of claim 8, wherein the filter system connects two filters and the movement of the piston causes the movement of one filter to be out of sync with the movement of the other filter.

10. The system of claim 7, wherein the piston further comprises a linear servo.

11. The system of claim 10, wherein the linear servo further comprises an encoder.

12. The system of claim 11, wherein the encoder provides information about a state of the system.

13. An alternating tangential flow pumping system comprising:

a first pneumatic cylinder having a proximal end, a distal end, and comprising a chamber, the proximal end connected in series to a first filter system, wherein the proximal end comprises an opening in the center of the proximal end, the opening leading to the chamber;

a second pneumatic cylinder having a proximal end, a distal end, and comprising a chamber, the proximal end serially connected to a second filter system, wherein the proximal end comprises an opening in the center of the proximal end, the opening leading to the chamber;

a piston connecting link entering each pneumatic cylinder through the distal end thereof;

the filter system, wherein the system comprises a hemisphere comprising a septum, the hemisphere connected to a distal end of a cylinder, wherein a proximal end of the cylinder is connected to a source of liquid, and the cylinder comprises a filter;

wherein the piston moves to provide pressure to the first filter to create an equal vacuum on the second filter.

14. A method of controlling an alternating tangential flow filtration process, the method comprising:

selecting the volume or size of the diaphragm pump;

selecting a volume of fluid discharged by the diaphragm pump over a given period of time;

selecting a duration of pumping operation that, together with a desired volume of fluid displaced over the duration, produces a performance curve suitable for a particular application;

the pumping operation is actuated by alternately supplying a positive and a negative air flow to the diaphragm pump using a linear servo.

15. The method of claim 14, wherein the linear servo comprises an encoder.

16. The method of claim 15, further comprising receiving a position signal from an encoder of the linear servo during at least one period of alternating positive and negative air flows.

17. The method of claim 16, further comprising comparing the position signal to a measured process variable and modifying an amplitude, duration or other characteristic of the period based on the comparison.

18. A method of actuating a diaphragm pump, comprising:

selecting the volume or size of the diaphragm pump;

selecting a volume of fluid discharged by the diaphragm pump over a given period of time;

selecting a duration of pumping operation that, together with a desired volume of fluid displaced over the duration, produces a performance curve suitable for a particular application;

actuating a pumping operation by alternately supplying a positive air flow and a negative air flow within a chamber connected to the diaphragm pump using a piston attached to a linear servo;

monitoring the movement of the linear servo to maintain or change the positive or negative air flow of the chamber per unit time;

optionally, monitoring the position of the diaphragm pump at one or both ends of its displacement; and

optionally, the positive or negative air flow within the chamber is alternated to affect the position of the diaphragm at one or both ends of its displacement, or to affect the duration of part of the displacement period or the entire displacement period, or both.

19. The method of claim 18, further comprising simultaneously filtering fluid drawn from the bioreactor through two filters, wherein the diaphragm pump activates both in a out of phase mode.

20. The method of claim 18, further comprising using the piston position to simulate a current state of the system.

21. The method of claim 20, further comprising using the piston position to calculate a transmembrane pressure in the system.

Technical Field

The present application relates generally to Alternating Tangential Flow (ATF) perfusion pumping methods, and more particularly to devices, systems, and methods of using the same.

Background

Filtration is typically performed to separate, clarify, modify and/or concentrate a fluid solution, mixture or suspension. In the biotechnology and pharmaceutical industries, filtration is critical to the successful production, handling and testing of new drugs, diagnostics and other biological products. For example, in the manufacture of biologicals, animal cell cultures are used, and filtration is performed to clarify, selectively remove and concentrate certain components from the culture medium, or to modify the culture medium prior to further processing. Filtration can also be used to increase productivity by maintaining perfusion cultures at high cell concentrations.

Filtration chemistry, construction and modes of use have been developed to facilitate the separation of materials based on their chemical and physical properties. Despite the extensive development of filtration technology, filters are often limited in their susceptibility to clogging. For example, when used to filter suspensions of cultured mammalian cells, they are easily clogged by dead cells, cell debris, aggregates, fiber biomolecules, or other components found in the complex "broth" of the culture. In this regard, the method of filtration can have a profound effect on filtration efficiency and membrane life. In one filtration process, commonly referred to as "dead-end" filtration, the entire fluid passes through the membrane perpendicular to the membrane surface. Debris quickly collects at the surface causing rapid clogging of the septum. Typically, applications using dead-end filtration involve small samples. The process is simple and relatively inexpensive. Another filtration process, commonly referred to as tangential flow filtration (also referred to as TFF), provides an improvement over dead-end filtration. In TFF, the fluid to be filtered is recirculated by a pump, typically from the reservoir through a filter and back to the reservoir. The flow through the filter is parallel to the surface of the filter. Effectively clearing any accumulation of debris by the "flushing" action of the circulating fluid; however, one of its limitations is the tendency to form gel-like deposits on the filter surface, which may limit the effectiveness of the filter and eventually block it. Another process known as alternating tangential flow filtration (ATF) provides yet another mode of filtration. ATF is similar to TFF, which creates a flow pattern parallel to the surface of the filtration membrane; however, it differs from TFF in that the direction of flow is repeatedly alternated or reversed across the filter surface. The alternating tangential flow filtration system described in U.S. patent No.6,544,424 to Shevitz, the entire contents of which are incorporated herein, includes a filter element (typically a hollow fiber cartridge) connected at one end to a reservoir containing the contents to be filtered and at the other end to a diaphragm pump capable of receiving and reversibly draining unfiltered liquid that reversibly flows between the reservoir and the pump through the filter element. The system shows the ability to maintain a filtered complex mixture comprising cell culture media even if the media is loaded with high cell concentrations and other cellular products. However, the range of applications of this system is limited.

There are a wide variety of filtration systems suitable for large scale filtration of media in a variety of applications. However, such systems require positive and negative pressure supplies. The positive and negative pressures may be provided by a facility in which the positive and negative pressures are shared by other users and result in a change in consistency due to distance from the pressure source. Positive and negative pressure may also be provided by a generator, which may be noisy and obtrusive in a laboratory environment. Current systems do not accurately adjust the duration of the transition between positive and negative airflow or the amount of airflow. Furthermore, current systems often involve many components in a complex assembly, which is difficult to maintain. On the other hand, embodiments of the present disclosure allow for precise control of the duration and amount of the transition, as described in more detail below.

Disclosure of Invention

Drawings

Figure 1 is an image of a filter attached to a cylinder with a piston.

Fig. 2A-B are schematic diagrams illustrating the position of a pressure and vacuum generating piston and the corresponding position of a diaphragm.

FIG. 3 is a schematic diagram showing dual filter activation coupled to the same air and vacuum generating cylinder, with the membrane operating in an out-of-plane mode.

Detailed Description

Overview

An Alternating Tangential Flow (ATF) pumping method is disclosed in which positive and negative pressures are generated in use. The method uses a pneumatic cylinder connected to a diaphragm pump of the ATF filter. The pneumatic cylinder contains a piston that allows controlled creation of positive and negative pressure on the diaphragm. The movement of the membrane allows fluid to enter and exit through the ATF filter. Fig. 1 illustrates an embodiment of the present disclosure. As shown in fig. 1, filter 100 is connected to cylinder 102 by base locking feature 101. The cylinder 102 is also connected to a linear servo via a connection 103.

The end of the cylinder without the piston connecting rod has an opening in the centre, which opens into the functional chamber of the cylinder. The bottom of the filter hemisphere base has an opening that matches the opening at the end of the cylinder. The cylinder face has a locking system that corresponds to a receiver on the filter to allow a secure connection between the filter and the pressure and vacuum sources. When connected, the openings in the pressure and vacuum sources are activated by a linear servo or an electric linear actuator.

Figure 2A illustrates one embodiment of a device with a septum 200a in a top position. Within the cylinder 204, a piston 210 generates a positive pressure by moving toward a filter 212. Fig. 2B shows an embodiment of the device in which the diaphragm 200B is in the bottom position and the piston 210 creates a negative pressure by moving downward toward the linear servo 206.

A linear servo or an electric linear actuator is connected to a piston which enters the cylinder through the end opposite to the end connected to the filter. As the piston moves away from the filter base, a vacuum is created and the cell culture is drawn into the filter housing. When the piston moves towards the filter, pressure is generated and the cell culture is pushed out of the filter housing.

The speed and control of the piston movement can be controlled by a linear servo or an electric linear actuator, which is in turn controlled by an algorithm dictated by a PLC or PC. To overcome the compressibility of air, a continuous, uniform pressure and vacuum was applied to the filter hemisphere base containing the diaphragm pump.

The linear servomechanism or the electric linear actuator is equipped with an encoder that allows the exact position of the piston to be known at all times. This allows the piston to move in either a full stroke or a partial stroke depending on the needs of the system. Thus, in systems using different sized filters, the piston system can be adjusted to provide the appropriate level of pressure or vacuum required.

In embodiments of the present disclosure, a single piston and cylinder may be connected to multiple ATF filter units to provide positive and negative pressures in parallel. For example, when two filters are in sequence, a cylinder may be attached to both filters such that when the piston moves to provide pressure to one filter, an equal vacuum is applied to the second filter, and vice versa. Fig. 3 shows an embodiment of the described multi-filter system 300. A positive/negative pressure is created in the cylinder 302, causing the diaphragm 306 to become out of sync. A linear servo 304 is attached to the cylinder 302.

The movement of the piston may be adjusted due to changing conditions within the system, such as changes in the viscosity of the pumped liquid.