WO2011038008A2

WO2011038008A2 - Method and apparatus for sterile sampling for gmp reactor applications

Info

Publication number: WO2011038008A2
Application number: PCT/US2010/049835
Authority: WO
Inventors: Jr. George E. Barringer
Original assignee: Groton Biosystems, Llc
Priority date: 2009-09-23
Filing date: 2010-09-22
Publication date: 2011-03-31
Also published as: WO2011038008A3

Abstract

A method of sampling from a manufacturing system includes coupling a sampling valve and a sampling vessel in series to a fluid sample source, sterilizing the sampling valve and the sampling vessel, supplying a sample from the fluid sample source through the sterile sampling valve to the sterile sampling vessel, closing the sampling valve to close fluid communication between the fluid sample source and the sampling vessel, and directing the sample from the sterile sample vessel to a processing system.

Description

METHOD AND APPARATUS FOR STERILE SAMPLING FOR GMP REACTOR APPLICATIONS

RELATED APPLICATION

This application claims the benefit of U.S. Provisional Application No. 61/245,066, filed on September 23, 2009.

The entire teachings of the above application are incorporated herein by reference.

BACKGROUND OF THE INVENTION

In bioreactor processes, maintaining a contamination-free environment is key. Whenever a bioprocess system is exposed to the external environment, it faces the risk of contamination by viruses, micro-organisms, and chemicals. Typical bioprocesses involve batch bioreactors where cells are cultured and harvested over a period of time ranging from hours to days. After a batch is harvested, the reactor vessel is sterilized in preparation for the next batch process. For small volume reactors, the entire reactor system can be placed in an autoclave and completely sterilized. For example, reactors that are about 5 liters or less typically are made of glass and are sterilized in an autoclave. However, large volume reactors, such as those that are about 5 liters or more, are typically too large to be placed in an autoclave, and must therefore be sanitized using Clean-in-Place (CIP) and sterilized using Steam-in-Place (SIP) methods. CIP and SIP are methods used in the pharmaceutical and food industries for the in-line sterilization of processing equipment, including vessels, valves, process lines, and filter assemblies. These methods are used to achieve sterility or a certain level of sanitation required by regulation for a particular process.

In many cases, bioreactor processes do not lend themselves easily to in-situ analysis of the batch. Instead, samples must be physically extracted from the process and examined and manipulated outside the vessel, thereby exposing the

1019633.1 entire batch to the external environment and the possibility of contamination. The devices and methods described in U.S. Application Nos. 12/490,960 of Barringer and 12/491,014 of Yu et al., both of which are incorporated herein by reference, provide for sanitary sampling of material from a reactor/vessel in an open system using a sequence of SIP and CIP procedures. As used herein, the term "open" refers to a system having a sterile environment that, during operation, comes into direct communication with a non-sterile environment. Many biological research and development processes where sterile environments can be exposed to sanitary environments are amenable to such open sampling systems, such as process research and development. The products of these processes are typically not used directly in patient care. However, some manufacturing processes that operate under Good Manufacturing Practice (GMP) procedures based on regulations promulgated by the US Food and Drug Administration require a greater degree of sterility within the system. That is, certain manufacturing systems are required to be "closed." As used herein, the term "closed" refers to a system having a sterile environment that, during operation, does not come into direct communication with a non-sterile environment. In such processes, a sampling system that communicates directly with the sterile reactor or vessel containing the material of interest must also be sterile.

Sampling within a closed system has conventionally been carried out manually by connecting a pre-sterilized sample container to the outlet of the reactor/vessel containing the sample of interest. For example, a pre-sterilized container sealed by a one-time use septa is placed in communication with the vessel by puncturing the septa with a needle in fluid communication with the vessel. The needle then injects the sample into the pre-sterilized container. The container is then removed from the system and the sample is analyzed.

SUMMARY OF THE INVENTION

What is needed is a device and method for automatically obtaining a sample from a sterile system without exposing said system directly to a non-sterile environment.

1019633.1 In one aspect, the present invention is directed to a method of sampling from a manufacturing system. The method includes coupling a sampling valve and a sampling vessel in series to a fluid sample source, sterilizing the sampling valve and the sampling vessel, supplying a sample from the fluid sample source through the sterile sampling valve to the sterile sampling vessel, closing the sampling valve to close fluid communication between the fluid sample source and the sampling vessel, and directing the sample from the sterile sample vessel to a processing system.

The method may further include the step of equalizing pressure between the sampling vessel and a headspace of the fluid sample source through a sterile headspace valve prior to supplying the sample to the sterile sampling vessel. The method may still further include the step of cooling the sterile sampling vessel prior to supplying the sample to the sterile sampling vessel. The step of cooling may include passing sterile water through the sterile sampling vessel. The step of cooling can include circulating cooling fluid through a cooling jacket in thermal

communication with the sterile sampling vessel and may include monitoring the temperature of the sampling vessel. The step of sterilizing can include passing steam through the sampling valve and the sampling vessel for a duration sufficient to sterilize the sampling valve, the sampling vessel, and the fluid path therebetween. The method may further include controlling a sterile air valve to allow sterile air to enter the sampling vessel and displace the sample to a location downstream of the sampling vessel. The method may still further include the step of flushing the sterile sampling vessel with sterile water after directing the sample from the sterile sampling vessel to the processing system. In an embodiment, the method further includes the step of monitoring the volume of the sample supplied to the sterile sampling vessel.

In another aspect, the present invention is directed to an automatic sterile sampling system for obtaining a sample within a manufacturing process. The system includes a steam valve configured to receive steam from a steam source. The system further includes a sampling valve configured to receive fluid from a fluid sample source and connected to receive the steam from the steam source. The system still further includes a sampling vessel connected to receive fluid samples

1019633.1 from the sampling valve and connected to receive steam from the steam valve. The sampling vessel is in fluid communication with a processing system. The system also includes a controller coupled to the steam valve to control the flow of the steam to sterilize the sampling valve and the sampling vessel prior to sampling and coupled to the sampling valve to control the flow of the fluid sample through the sampling valve to the sampling vessel.

The system may further include a steam source to which the steam valve is coupled. In addition, the system may include a closed system fluid sampling source to which the sampling valve is coupled. The system may further include a headspace valve configured to couple a headspace of the fluid sample source with the sampling vessel. In addition, the controller may be further coupled to the headspace valve to control equalization of pressure between the sampling vessel and the headspace of the fluid sample source. In an embodiment, the system includes a sterile air valve connected to receive sterile air from a sterile air source, the sterile air valve being in fluid communication with the sampling vessel. Further, the controller may be coupled to the sterile air valve to control the flow of sterile air into the sampling vessel to equilibrate pressure in the sampling vessel with components located downstream of the sampling vessel. The controller may also be coupled to the sterile air valve to control the flow of sterile air into the sampling vessel to displace the fluid sample to a location downstream of the sampling vessel. The system may further include a sterile water valve connected to receive sterile water from a sterile water source, the sterile water valve being in fluid communication with the sampling vessel. . Further, the controller may be coupled to the sterile water valve to control the flow of the sterile water. The system may further include a drain valve connected to a drain to pass the steam from the sampling vessel to the drain and to pass the fluid sample from the sampling vessel to the drain. In addition, the controller may be coupled to the drain valve to control the passing of the steam to the drain and the passing of the fluid sample to the drain. Further still, the system may include an isolation valve connected to receive the fluid sample form the sampling vessel and to pass the fluid sample to the processing system. In addition, the controller may be coupled to the isolation valve to control the passing of the

1019633.1 fluid sample to the processing system. In an embodiment, the system includes a manual sampling valve connected to receive the fluid sample from the sampling vessel and pass the fluid sample to a manual sampling output port. In an embodiment, the system includes a cooling jacket connected to receive cooling fluid from a cooling fluid source, the cooling jacked being in thermal communication with the sampling vessel.

BRIEF DESCRIPTION OF THE DRAWING

The foregoing will be apparent from the following more particular description of example embodiments of the invention, as illustrated in the accompanying drawings in which like reference characters refer to the same parts throughout the different views. The drawings are not necessarily to scale, emphasis instead being placed upon illustrating embodiments of the present invention.

FIG. 1 is a drawing of an embodiment of an automated system at rest according to the present invention;

FIG. 2 is a drawing of an alternative embodiment of the automated system at rest;

FIG. 3 is a drawing for the control valve system for the isolation valve; FIG. 4 is a drawing of the automated system during a sterilization operation; FIG. 5 is a drawing of the automated system during a cooling operation; FIG. 6 is a drawing of the automated system during a cooling operation;

FIG. 7 is a drawing of the automated system during a sample extraction operation;

FIG. 8 is a drawing of the automated system during a sample transfer operation;

FIG. 9 is a drawing of the automated system during a sample transfer operation;

FIG. 10 is a drawing of a sampling valve during a sterilization operation; FIG. 11 is a drawing of a sampling valve during a sampling operation.

DETAILED DESCRIPTION OF THE INVENTION

1019633.1 A description of example embodiments of the invention follows. The embodiments provide an automated sampling system within a closed manufacturing system and method for sampling from a closed manufacturing system while ensuring that the manufacturing system, during operation, does not come into direct communication with a non-sterile environment. For example, the sampling system may be configured to obtain samples from a component in a closed system, such as a bioreactor. The invention is not limited to sampling from a bioreactor, but rather can be applied to the sterile sampling of any vessel or other sample source containing a fluid. The system employs a series of pneumatically or electrically actuated valves to control the flow of steam, fluid sample, and optionally air and/or cooling fluid through the system at specified times and includes a sample vessel through which fluid samples are routed from the bioreactor vessel to a downstream processing system. As used herein, the term "valve" refers to a single valve or system of valving that achieves a particular flow configuration. As used herein, "fluid communication" refers to a relationship between two components by which fluid can be permitted to flow from one component to the other.

Referring to the FIG. 1 , a sample source, such as reactor vessel 1 is fluidly connected to sampling valve 3. Samples from sample source 1 are dispensed through sampling valve 3 and directed into sample vessel 61. Sample vessel 61 is approximately the same form factor (ratio of height and diameter) as the reactor vessel. It is believed that using a sampling vessel that has the same form factor as the reactor vessel achieves a constant surface area to volume ratio and headspace to liquid volume ratio. It is further believed that keeping these ratios constant helps to achieve equivalent gas saturation in the fluid of both the reactor vessel and sample vessel. For reasons described further below, such equivalent gas saturation is desirable. Temperature sensor 65 and level sensor 67 at sample vessel 61 provide for monitoring of the sample in the sample vessel. Sample vessel 61 is in thermal communication with cooling jacket 63, which circulates cooling fluid from cooling fluid source 75.

As shown in FIG. 1, fluid channel 4 establishes fluid communication between sampling vessel 61 and manual sample sampling valve 15, isolation valve

1019633.1 17, and drain valve 19. Samples exit sample vessel 61 through fluid channel 4 and can be directed to various locations downstream. For example, manual sampling valve 15 may be opened and sample fluid may pass through manual sampling output port 21 if the user wishes to obtain samples manually. For automated sampling, isolation valve 17 can be controlled to allow fluid samples to pass from fluid channel 4 to sample transfer channel 6. These samples are directed into sampling processing system 11 for automated processing and analysis of the sample.

Processing system 11 can include cleaning, processing, and analytical

instrumentation, as well as controller 27. An example of a suitable processing system is described in U.S. Patent Application Publication No. 2004/0259266, incorporated herein by reference in its entirety. Alternatively, the sample may also be discarded by opening drain valve 19 to route the sample through drain channel 8 to drain 10.

Referring back to FIG. 1, the top of sample vessel 61 is connected to the headspace 71 of reactor vessel 1 by way of headspace pipe 73. Headspace pipe 73 may be fitted optionally with sterile filters, fluid traps, or other accessories.

Headspace pipe 73 is fitted with an automatic valve 81, which controls the flow of headspace gas and provides for pressure equilibrium between reactor vessel 1 and sample vessel 61. Equilibrating or equalizing the pressure between reactor vessel headspace 71 and sample vessel 61 prevents outgassing of dissolved gas

components from fluid samples transferred from the reactor vessel 1 to the sampling vessel 61 and achieves equivalent gas saturation in the fluid of both the reactor vessel and sample vessel. Thus, the concentration of components in the sample does not change as a result of introduction into sample vessel 61, and the sample in sampling vessel 61 is representative of the fluid in the reactor vessel 1.

Headspace pipe 73 is connected to steam source 5, optional sterile air source 77, and optional sterile water source 117. Steam source 5 is provided for sterilization of the system, particularly sampling valve 3 and sampling vessel 61. Steam is allowed to pass through steam valve 13 into headspace pipe 73.

Headspace valve 81 is a three-port valve that includes one valved leg, which can be open or closed, and two common legs, which are in fluid communication.

1019633.1 The valved leg of headspace valve 81 is in fluid communication with headspace 71. When headspace valve 81 is open, headspace 71 is in fluid communication with sampling vessel 61, steam valve 13, and steam channel 2. Preferably steam valve 13 and sampling valve 3 are located in close proximity to headspace valve 81, because these valves will be in contact with the sample.

Steam valve 13 preferably is in close proximity to head space valve 81, in order to minimize "dead legs," i.e., tubing not directly contacted by steam.

Preferably, all tubing connections in contact with the sample are minimized. The term sample here refers to the contents of the reactor vessel 1 , whether in liquid phase (bottom of reactor) or gas phase (top of reactor). For example, the following tubing connections or fluid paths may be minimized: from headspace pipe 73 to headspace valve 81, from headspace valve 81 to sampling vessel 61, from headspace 71 to sampling valve 3, from sampling valve 3 to headspace pipe 73, from steam valve 13 to headspace valve 81, from sterile air valve 121 to headspace pipe 73, from sterile water valve 119 to headspace pipe 73, from fluid channel 4 to manual sampling valve 15, from fluid channel 4 to isolation valve 17, from fluid channel 4 to drain valve 19, and fluid channel 4.

Steam source 5 is also fluidly connected to sampling valve 3 and sample vessel 61, and is therefore capable of sterilizing both components. Steam passes through sampling valve 3 and sampling vessel 61 and exits to drain 10.

Sample vessel 61 may be connected to sterile air source 77 to equilibrate pressure in sample vessel 61 with components downstream of sample vessel 61. The connection is automatically controlled by manipulating an optional sterile air valve 121 between sample vessel 61 and sterile air source 77. When a sample is released from sample vessel 61 to a downstream location, sterile air valve allows sterile air to enter sample vessel 61 and displace the fluid sample downstream. The sterile air source 77 may also be used to cool the sample vessel or may be used to blow down steam condensate into steam drain 10 after the sample vessel 61 has cooled and before the sample is admitted from the reactor vessel 1 to the sample vessel, thereby eliminating liquid that could potentially dilute the next sample to enter sample vessel 61.

1019633.1 Sample vessel 61 may be also connected to sterile water source 117. Sterile water valve 119 is controlled to allow sterile water to pass through sampling vessel 61 to reduce the temperature of sampling vessel 61 after steam sterilization of sampling vessel 61. In addition, sterile water from sterile water valve 119 can be used to flush residual fluid from the sampling vessel prior to sterilization in preparation for a new sample.

Steam valve 13 controls the flow of steam through a steam channel 2. Steam valve 13 is typically a diaphragm valve, such as GEMU^® Type 650/015/D80415 A0- 1537, which is a 1/2 inch two-port pneumatically actuated sanitary valve. When steam valve 13 is open, steam is allowed to pass through steam channel 2 to sampling valve 3.

Sampling valve 3 is typically a three-port plunger valve specifically adapted for sterile sampling of a liquid sample from a container, such as the KEOFITT^®

Wl 5 sampling valve, or the valves described in U.S. Patent Application

Publication No. 2007/0074761 incorporated herein by reference in its entirety. An example of a suitable KEOFITT^® sampling valve is shown in FIGS. 10 and 11. Sampling valve 3 is connected to three components of the system: the steam channel 2, a fluid sample source 1, such as a reactor vessel, and a steam/sample channel 4. Steam and fluid samples can flow from the sampling valve 3 to sampling vessel 61 through steam/sample channel 4. Steam/sample channel 4 typically has an inner diameter of about 9 mm. When the sampling valve 3 is closed as shown in FIG. 10, steam is able to flow from steam channel 2 to steam/sample channel 4. When opened, as shown in FIG. 11, fluid sample flows from port 33 toward steam/sample channel 4, the flow path toward steam channel 2 being blocked by steam valve 13.

Isolation valve 17 is typically a two-port diaphragm valve, such as a GEMU^®

Type 650 TC TFE 15RaEP Conl having two 3/8 inch (9 mm nominal) ports, which is a pneumatically actuated sanitary valve.

Drain valve 19 is typically similar to isolation valve 17. An example of a suitable drain valve is a GEMTj^® Type 650 TC TFE 15RaEP Conl having two 3/8 inch (9 mm nominal) ports, which is a pneumatically actuated sanitary valve.

1019633.1 Manual sampling valve 15 is typically a two-port valve, such as GEMU^® Type 650 TC TFE 15RaEP Conl with manual actuator. Manual sampling valve 15 is connected to manual sampling output port 21, which can be used by a human operator to draw fluid samples from the sampling vessel 61. The manual valve operates in a similar manner as the isolation valve 17. However, during normal automatic operation, the valve shuts the fluid pathway to manual output port 21.

Headspace valve 81 is typically a three-port diaphragm valve, such as a GEMU^® Type 650 TC TFE 15RaEP Con A-B, which is a 3/8 inch three-port pneumatically actuated sanitary valve.

Sterile water valve 119 and sterile air valve 121 are typically two-port valves, such as GEMU^® Type 650 TC TFE 15RaEP Conl with pneumatic actuator or equivalent.

The steam valve 13, sampling valve 3, isolation valve 17, drain valve 19, headspace valve 81, manual sampling valve 15, optional sterile water valve 119, and optional sterile air valve 121 are controlled in sequence to perform various system operations, which will be described in detail below. Each of the valves may be pneumatically actuated by one of two control valves in parallel: a solenoid control valve and a manual control valve. For example, FIG. 3 shows isolation valve 17, which is pneumatically actuated by either manual control valve 36 or solenoid control valve 35. The user can select between automatic and manual control by toggling auto/manual solenoid switch valve 34, which is connected to compressed air source 33. The valve switches compressed air from source 33 to either the solenoid valve 35 for automatic control or manual control valve 36 for manual control. Under normal operation, the valves of the system are controlled automatically. A controller 27, such as a programmable logic controller (PLC) controls the solenoid valves and solenoid switch valves. As shown in FIG. 1, the controller typically resides in processing system 1 1 and controls the control valves to actuate the pneumatic valves, thereby automatically performing the various operations of the system in sequential order periodically throughout the bioreactor process.

1019633.1 The downstream set-up as shown in FIG. 1 can be alternatively achieved by using three-port or three-way valves, such as shown in FIG. 2 and as described in U.S. Application No. 12/490,960, which is incorporated herein by reference. As shown in FIG. 2, manual sampling valve 15 and isolation valve 17 are three-port valves. Fluid channel 4' establishes fluid communication between sampling vessel 61 and manual sampling valve 15. Sampling vessel 61 is connected to isolation valve 17 through manual sampling valve 15 and further connected to drain valve 19 through isolation valve 17.

In the system shown in FIG. 2, isolation valve 17 may be a three-port diaphragm valve. An example of a suitable isolation valve is a GEMlj^® Type 650 TC TFE 15RaEP Con A-B, which is a 3/8 inch three-port pneumatically actuated sanitary valve. A first port of isolation valve 17 is connected to the steam/fluid channel 4', while a second port of isolation valve 17 is connected to drain channel 8 and a third port of the isolation valve 17 is connected to sample transfer channel 6.

In the system shown in FIG. 2, manual sampling valve 15 is typically a three-port diaphragm valve, such as GEMIJ^® Type 601 TC TFE 15RaEP Con A-B, which is a 3/8 inch three-port manually actuated sanitary valve.

During the sampling operation, controller 27 automatically manipulates the system. As shown in the FIG. 1, controller 27 is connected to headspace valve 81, steam valve 13, sterile air valve 121, sterile water valve 119, sampling valve 3, isolation valve 17, manual sampling valve 15, and drain valve 19. The controller may be also connected to temperature sensor 65, level sensor 67, and cooling fluid source 75.

In one embodiment, the controller operates the sampling system in the following sequence.

Steam Sterilization

FIG. 4 is a drawing of the automated reactor sampling system during a sterilization operation for the sampling system when the reactor is in operation. The sampling process begins by first sterilizing sampling valve 3 and sampling vessel 61. As shown in FIG. 4, steam valve 13 and drain valve 19 are opened, allowing steam source 5 to provide steam through sampling valve 3 and sampling vessel 61 to

1019633.1 drain 10. Steam is typically allowed to pass through sampling valve 3 and sampling vessel 61 for about 20-25 minutes at 121 °C or higher to ensure complete

sterilization of these components. During this time, reactor sampling valve 3, manual sampling valve 15, and isolation valve 17 are closed, so that steam cannot pass to reactor vessel 1, manual sampling output port 21, or sample transfer channel 6. During sterilization when the reactor is in operation, headspace valve 81 , and optional sterile air and sterile water valves 121, 119 are also closed. The steam also sterilizes headspace valve 81. After sterilization, steam valve 13 is closed.

For preparation of a reactor to be put in use, a preliminary sterilization must be performed. In a preliminary operation before putting the reactor in service for a fermentation cycle the entire reactor system, including headspace pipe 73 and headspace 71, is steam sterilized in addition to the steam path shown in FIG. 4. In that case, headspace valve 81 is open and steam can pass through headspace pipe 73, headspace 71, and the fluid path from headspace 71 to sampling valve 3.

FIG. 10 shows the sterilizing of a sampling valve, such as sampling valve 3, in detail. Valve head 31 is seated over an aperture 33, thereby obstructing the flow of fluid from a fluid sample source, such as reactor vessel 1. Steam enters sampling valve 3 from the steam source (not shown) through steam channel 2 and exits through steam/sample channel 4. Headspace valve 81 and headspace pipe 73 are not shown in FIG. 10.

Cool-down

After the sampling vessel is sterilized, it must be cooled, so that incoming fluid samples are not denatured or otherwise damaged as a result of residual heat from the sterilization operation. As shown in FIG. 5 sampling vessel 61 may be cooled by sterile water from sterile water source 117. For example, sterile water valve 119 is opened and sterile water is allowed to pass through sampling vessel 61 until the vessel reaches a desired temperature. The temperature of sample vessel 61 may be monitored by temperature sensor 65. Drain valve 19 remains open from the sterilization operation, and the sterile water exits the system by way of drain 10. At the end of the cooling operation, sterile water valve 119 and drain valve 19 are closed. Alternatively, as shown in FIG. 6, drain valve 19 may be closed prior to the

1019633.1. cooling operation, and sample vessel 61 is cooled by circulating cooling fluid from cooling fluid source 75 through cooling jacket 63, which is in thermal

communication with sterile sampling vessel 61.

Sample Extraction

After sampling vessel 61 is cool enough to handle fluid samples, a fluid sample is ready to be extracted from reactor vessel 1. FIG. 7 is a drawing of the automated system during a sampling operation. First, headspace valve 81 is opened, followed by reactor sample valve 3 after a brief delay. Fluid from reactor vessel 1 is allowed to enter sampling vessel 61 through sampling valve 3 until the desired volume of sample is obtained in sampling vessel 61. Because headspace valve 81 remains open during sample extraction, any dissolved gas components in the sample remain saturated in the fluid. Thus, the concentration of the fluid sample after extraction from reactor vessel 1 is unchanged with respect to the concentration of the same fluid sample prior to extraction from reactor vessel 1. The volume of sample entering sampling vessel 61 may be monitored by level sensor 67. Once the desired amount of sample is obtained, sampling valve 3 and headspace valve 81 are closed.

FIG. 11 shows as ample valve, such as sampling valve 3, during the sampling operation in detail. Valve head 31 is removed from port 33 by pneumatic control and fluid is allowed to flow from a fluid sample source, such as fluid sample source I, through steam/sample channel 4. The steam valve 13 along the steam channel 2 prevents fluid samples from flowing to the steam source. Headspace valve 81 and headspace pipe 73 are not shown in FIG. 11.

Sample Transfer

The sample is now contained in sample vessel 61 and fluid communication to reactor vessel is closed. Now, the sample may be transferred downstream without danger of exposing reactor vessel 1 to a non-sterile environment. The sample is allowed to exit sample vessel 61 and can be directed to one of several parts of the system, as described above. For example, isolation valve 17 can be opened to allow the sample to pass through sample transfer channel 6 into automated processing

1019633.1 system 11 as shown in FIG. 8. Alternatively, the sample can be obtained manually by opening manual samplmg valve 15 to allow the sample to pass through manual sampling output port 21 as shown in FIG. 9. On the other hand, if the operator wishes to discard the sample, drain valve 19 can be opened. After transfer of the sample from sample vessel 61 to the desired downstream system component, the respective valve leading thereto is closed. Sterile air valve 121 may be controlled to allow sterile air to enter sample vessel 61 and displace the fluid sample downstream.

Flush and Sterilize

With the sample successfully transferred, the system is again sterilized. First, sterile water valve 119 is opened and sterile water is allowed to pass through sample vessel 61 to flush the system of any residual sample fluid. Drain valve 19 is opened, and the sterile water exits the system by way of drain 10. This operation is similar to cooling the sampling vessel with sterile water as described with reference to FIG. 5. After the system is sufficiently flushed, sterile water valve 119 and drain valve 19 are closed. Steam valve 13 and drain valve 19 are opened, allowing steam source 5 to provide steam through sampling valve 3 and sampling vessel 61 to drain 10. This operation is similar to steam sterilization described with reference to FIG. 4. After sterilization, steam valve 13 is closed. Sample vessel 61 may be cooled by sterile water from sterile water source 117 or by circulating cooling fluid from cooling fluid source 75 through cooling jacket 63.

The teachings of all patents, published applications and references cited herein are incorporated by reference in their entirety. While this invention has been particularly shown and described with references to example embodiments thereof, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the scope of the invention encompassed by the appended claims.

1019633.1

Claims

CLAIMS What is claimed is:

1. A method of sampling from a manufacturing system, comprising:

coupling a sampling valve and a sampling vessel in series to a fluid sample source;

sterilizing the sampling valve and the sampling vessel;

supplying a sample from the fluid sample source through the sterile sampling valve to the sterile sampling vessel;

closing the sampling valve to close fluid communication between the fluid sample source and the sampling vessel; and

directing the sample from the sterile sampling vessel to a processing system.

2. The method of claim 1 , further comprising the step of equalizing pressure between the sampling vessel and a headspace of the fluid sample source through a sterile headspace valve prior to supplying the sample to the sterile sampling vessel.

3. The method of claim 1, further comprising the step of cooling the sterile sampling vessel prior to supplying the sample to the sterile sampling vessel.

4. The method of claim 3, wherein the step of cooling comprises passing sterile water through the sterile sampling vessel.

5. The method of claim 3, wherein the step of cooling comprises circulating cooling fluid through a cooling jacket in thermal communication with the sterile sampling vessel.

6. The method of claim 3, wherein the step of cooling comprises monitoring the temperature of the sampling vessel.

1019633.1

7. The method of claim 1, wherein the step of sterilizing comprises passing steam through the sampling valve and the sampling vessel for a duration sufficient to sterilize the sampling valve, the sampling vessel, and the fluid path therebetween.

8. The method of claim 1, further comprising controlling a sterile air valve to allow sterile air to enter the sampling vessel and displace the sample to a location downstream of the sampling vessel.

9. The method of claim 1, further comprising the step of flushing the sterile sampling vessel with sterile water after directing the sample from the sterile sampling vessel to the processing system.

10. The method of claim 1 , further comprising the step of monitoring the volume of the sample supplied to the sterile sampling vessel.

11. An automatic sterile sampling system for obtaining a sample within a manufacturing process, comprising:

a steam valve configured to receive steam from a steam source;

a sampling valve configured to receive fluid from a fluid sample source and connected to receive the steam from the steam source;

a sampling vessel connected to receive fluid samples from the sampling valve and connected to receive steam from the steam valve, the sampling vessel being in fluid communication with a processing system; and

a controller coupled to the steam valve to control the flow of the steam to sterilize the sampling valve and the sampling vessel prior to sampling and coupled to the sampling valve to control the flow of the fluid sample through the sampling valve to the sampling vessel.

12. The system of claim 11, further comprising a steam source to which the steam valve is coupled.

1019633.1

13. The system of claim 11, further comprising a closed system fluid sample source to which the sampling valve is coupled.

14. The system of claim 11 , further comprising a headspace valve, configured to couple a headspace of the fluid sample source with the sampling vessel, and wherein the controller is further coupled to the headspace valve to control equalization of pressure between the sampling vessel and the headspace of the fluid sample source.

15. The system of claim 11, further comprising a sterile air valve connected to receive sterile air from a sterile air source, the sterile air valve being in fluid communication with the sampling vessel.

16. The system of claim 15, wherein the controller is further coupled to the sterile air valve to control the flow of sterile air into the sampling vessel to equilibrate pressure in the sampling vessel with components located downstream of the sampling vessel.

17. The system of claim 15, wherein the controller is further coupled to the sterile air valve to control the flow of sterile air into the sampling vessel to displace the fluid sample to a location downstream of the sampling vessel.

18. The system of claim 11, further comprising a sterile water valve connected to receive sterile water from a sterile water source, the sterile water valve being in fluid communication with the sampling vessel, and wherein the controller is further coupled to the sterile water valve to control the flow of the sterile water.

19. The system of claim 11, further comprising a drain valve connected to a drain to pass the steam from the sampling vessel to the drain and to pass the fluid sample from the sampling vessel to the drain, and wherein the controller is further coupled to the drain valve to control the passing of the steam to the drain and the passing of the fluid sample to the drain.

1019633.1

20. The system of claim 1 1, further comprising an isolation valve connected to receive the fluid sample from the sampling vessel and to pass the fluid sample to the processing system, and wherein the controller is further coupled to the isolation valve to control the passing of the fluid sample to the processing system.

21. The system of claim 11 , further comprising a manual sampling valve connected to receive the fluid sample from the sampling vessel and pass the fluid sample to a manual sampling output port.

22. The system of claim 11 , further comprising a cooling jacket connected to receive cooling fluid from a cooling fluid source, the cooling jacked being in thermal communication with the sampling vessel.

1019633.1