Patent Publication Number: US-11644349-B2

Title: Probe assembly

Description:
RELATED APPLICATIONS 
     This application is a continuation of U.S. patent application Ser. No. 15/861,201, filed on Jan. 3, 2018, now U.S. Pat. No. 10,859,412, which is a continuation of U.S. patent application Ser. No. 14/353,344 filed Apr. 22, 2014, now U.S. Pat. No. 9,891,080, issued Feb. 13, 2018, which is a filing under 35 U.S.C. 371 of international application number PCT/US2012/062345, filed on Oct. 28, 2012, published May 2, 2013 as WO 2013/063550, which claims the benefit of U.S. Provisional Patent Application No. 61/552,974, filed on Oct. 28, 2011, and entitled “Probe Assembly,” the teachings of all of the above referenced applications being incorporated herein by reference in their entirety. 
    
    
     FIELD OF INVENTION 
     This disclosure relates generally to bioprocessing systems and methods and, in particular, to systems and methods for inserting sensors into bioreactor vessels and tubing, including flexible or semi-rigid bags or tubing. 
     BACKGROUND 
     A variety of vessels, devices, components and unit operations are known for carrying out biochemical and/or biological processes and/or manipulating liquids and other products of such processes. Increasingly, in order to avoid the time, expense, and difficulties associated with sterilizing the vessels used in biopharmaceutical manufacturing processes, single-use or disposable bioreactor bags and single-use mixer bags are used as such vessels. For instance, biological materials (e.g., animal and plant cells) including, for example, mammalian, plant or insect cells and microbial cultures can be processed using disposable or single-use mixers and bioreactors. 
     The manufacturing of complex biological products such as proteins (e.g., monoclonal antibodies, peptides, hormones, and vaccine immunogens) requires, in many instances, multiple processing steps ranging from cell culture (bacteria, yeast, insect, fungi, etc.) and/or fermentation, to primary recovery, purification, and others. Conventional bioreactor-based manufacturing of biological products generally utilizes batch, or fed-batch processing through a series of unit operations with subsequent off-line laboratory analysis conducted on representative samples collected from various points of the process to ensure quality. 
     In order to obtain timely information regarding changing conditions within a bioreactor vessel during its operation, the use of sensor technology has been employed. With regard to use of disposable bioreactors, there are recognized difficulties in sterilely inserting a sensor into a flexible-walled bioreactor or flexible tubing that feeds or drains such vessels. Further, optical, electrical, and pH sensors, for example, positioned inside a flexible bag or tubing require an attachment means that allows for a clear signal to be communicated to or from external analytical instrumentation. Thus, there is an ongoing need for an improved sensor connector and a method for inserting a sensor into flexible disposable bioreactor bags or fluid circulating tubing. 
     An improved device and method for sterilely inserting a non-disposable sensor or a disposable sensor into a flexible bioreactor bag or tubing would also be beneficial for use in bioreactor-based manufacturing systems that include in-line sensing in order to provide real-time data. 
     Because the sensor itself can be expensive, there is an on-going need for an improved device and method for sterilely inserting a sensor into a flexible bag or tubing, a device and method that facilitate the removal of the sensor from the disposable bag or tubing without damaging the sensor. With such an improved device and method, the bag or tubing can be discarded along with the sensor, or alternatively the sensor can be removed, re-sterilized, and re-used. 
     SUMMARY OF THE INVENTION 
     In accordance with the present invention there is provided a probe assembly for inserting a disposable or non-disposable sensor into a flexible bag or a semi-rigid vessel or tubing, the assembly including a distal, preferably aseptic, connector for coupling to the vessel or tubing (e.g., via a mating aseptic connector), a probe sheath comprising at least a portion that is rigid, the probe sheath extending longitudinally from the aseptic connector and having at least one inner longitudinal lumen configured to receive an elongate probe body and to permit longitudinal movement of the probe body within the probe sheath lumen, and an actuator for deploying a probe within the vessel or tubing by advancing the probe body through the aseptic connector to a position where the probe can measure at least one parameter within the vessel or tubing. In one embodiment of the invention, the entire probe sheath is rigid. 
     In one embodiment of the invention, the probe sheath comprises at least a portion that is non-collapsible and the inner longitudinal lumen is configured to sealably receive an elongate probe body and to permit longitudinal movement of the probe body within the probe sheath lumen. 
     Methods of aseptically inserting a probe into a flexible or semi-rigid vessel or tubing are also disclosed. Such methods can include the steps of (1) providing a probe assembly having a distal aseptic connector and a probe sheath extending longitudinally from the aseptic connector and having at least one inner longitudinal lumen configured to receive an elongate probe body, (2) connecting the probe assembly to a port associated with the vessel or tubing (e.g. via a mating disposable aseptic connector) and (3) inserting an elongate probe through a lumen in the probe sheath and advancing the probe through the lumen until at least a sensing portion of the elongate probe is aseptically disposed within the vessel or tubing (or otherwise in a position where the probe can measure at least one parameter within the vessel or tubing). 
     Another embodiment of the invention is a method of forming a probe insertion device and inserting it into a flexible-walled container or a tubing system, the method including: partially inserting a probe body into a first end of a first flexible tubing section; attaching a first end of a second flexible tubing section to a tubing port of the flexible-walled container or the tubing system; welding a second end of the first flexible tubing section to the second end of the second flexible tubing section, thereby forming a welded flexible tubing; and advancing the probe body through the welded flexible tubing and partially into the flexible-walled container or the tubing system, while allowing the first flexible tubing section of the welded flexible tubing to fold back upon itself, thereby forming a probe insertion device and inserting it into a flexible-walled container or tubing system. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
       The foregoing and other objects, features and advantages of the invention will be apparent from the following more particular description of illustrative 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 the principles of the invention. Those skilled in the art will understand that the devices and methods specifically described herein and illustrated in the accompanying drawings are non-limiting exemplary embodiments. The features illustrated or described in connection with one exemplary embodiment can be combined with the features of other embodiments. Such modifications and variations are intended to be included within the scope of the present invention. 
         FIG.  1 A  is a cross-sectional side view of an exemplary, plunger-type probe assembly according to the invention in a detached position prior to coupling with a bag or tubing. 
         FIG.  1 B  is another cross-sectional side view of the probe assembly of  FIG.  1 A  in a coupled position, joined to a bag or tubing. 
         FIG.  1 C  is a cross-sectional side view of the probe assembly of another embodiment according to the invention of a plunger-type probe assembly in a detached position prior to coupling with a bag or tubing, wherein the probe body includes a connector plug. 
         FIG.  1 D  is a cross-sectional side view of the probe assembly shown in  FIG.  1 C , wherein the probe assembly is in a coupled position, joined to a bag or tubing. 
         FIG.  2 A  is a cross-sectional side view of another embodiment of a probe assembly according to the invention (with telescoping elements) in a detached position prior to coupling with a bag or tubing. 
         FIG.  2 B  is a cross-sectional side view of the probe assembly of  FIG.  2 A  in a coupled position, joined to the bag or tubing. 
         FIG.  3 A  is a cross-sectional side view of yet another embodiment of a probe assembly according to the invention (with a balloon element) in an attached position but prior to probe insertion into a bag or tubing. 
         FIG.  3 B  is a cross-sectional side view of the probe assembly of  FIG.  3 A  in a coupled position with the probe inserted into the bag or tubing. 
         FIG.  4 A  is a cross-sectional side view of yet another embodiment of an aseptic connector device according to the invention in a position that is prior to welding a section (Part “i”) thereof to a section (Part “ii”) of the tubing that is shown in  FIG.  4 B , the tubing attached to a flexible walled container or tubing. 
         FIG.  5 A  shows a cross-sectional side view of the aseptic connector device of  FIG.  4 A  and the tubing shown in  FIG.  4 B  positioned within a tubing welder. 
         FIG.  5 B  shows Part “i” of the aseptic connector device and Part “ii” of the tubing welded together to form a probe assembly according to an embodiment of the invention. 
         FIG.  5 C  shows the probe assembly with the probe body in a position after it has been pushed into the container or tubing system according to an embodiment of the invention. 
         FIG.  5 D  shows the system of  FIG.  5 C  including an outer, perimeter hose clamp securing the probe in position. 
     
    
    
     DETAILED DESCRIPTION 
     As used herein, the terms “comprises,” “comprising,” “includes,” “including,” “has,” “having,” or any other variation thereof, are intended to cover a non-exclusive inclusion. For example, a process, article, or apparatus that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such process, article, or apparatus. Further, unless expressly stated to the contrary, “or” refers to an inclusive or and not to an exclusive “or.” 
     Additionally, any examples or illustrations given herein are not to be regarded in any way as restrictions on, limits to, or express definitions of, any term or terms with which they are utilized. Instead, these examples or illustrations are to be regarded as being described with respect to one particular embodiment and as illustrative only. Those of ordinary skill in the art will appreciate that any term or terms with which these examples or illustrations are utilized will encompass other embodiments that may or may not be given therewith or elsewhere in the specification and all such embodiments are intended to be included within the scope of that term or terms. Language designating such non-limiting examples and illustrations includes, but is not limited to: “for example,” “for instance,” “e.g.,” and “in one embodiment.” 
     The term “flexible,” as used herein, refers to a structure or material that is pliable, or capable of being bent without breaking, and may also refer to a material that is compressible or expandable. An example of a flexible structure is a bag formed of polyethylene film. The terms “rigid” and “semi-rigid” are used herein interchangeably to describe structures that are “non-collapsible,” that is to say structures that do not fold, collapse, or otherwise deform under normal forces to substantially reduce their elongate dimension. “Collapsible” is defined to include substantially flexible material that will fold onto or into itself, such as, for example, fabrics and materials that form “accordion-like” structures in response to a compressive force. Depending on the context, “semi-rigid” can also denote a structure that is more flexible than a “rigid” element, e.g., a bendable tube or conduit, but still one that does not collapse longitudinally under normal conditions and forces. 
     A “vessel” as the term is used herein, means a flexible bag, a flexible container, a semi-rigid container, or a flexible or semi-rigid tubing, as the case may be. The term “vessel” as used herein is intended to encompass bioreactor vessels having a wall or a portion of a wall that is flexible or semi-rigid, single use flexible bags, as well as other containers or conduits commonly used in biological or biochemical processing, including, for example, cell culture/purification systems, mixing systems, media/buffer preparation systems, and filtration/purification systems, e.g., chromatography and tangential flow filter systems, and their associated flow paths. As used herein, the term “bag” means a flexible or semi-rigid container, vessel, or tubing. 
     Typically a flexible bag used for mixing or bioprocessing is supported by a rigid support structure or supported within a rigid vessel. A probe assembly according to an embodiment of the invention is particularly useful for attaching to a disposable or single use flexible bioreactor or mixer bag, or a flexible tubing. Sterilizing a probe before it is inserted into a reactor bag or vessel is often essential. When the probe is inserted via a probe assembly, it may be necessary to sterilize the entire probe assembly, including any sheaths, connectors, and tubes, as well as the probe itself, prior to inserting the probe into the reactor vessel. Common methods of sterilization include, but are not limited to, autoclaving, radiation treatment, and chemical treatment. When an autoclave is used, it can be important for steam to reach all of the interior surfaces of a probe assembly, as well as the exterior portions. 
     A typical industry standard size sensor is about 12 mm diameter.times.225 mm long, but any size sensor can be used. The sensor can be installed as an elongate probe body that is configured to be advanced into the vessel via a probe sheath. This particularly advantageous when the vessel has a flexible or non-rigid form. An aseptic connector is commonly used to perform the sterile connection between the probe sheath assembly and the sterile vessel. 
     Aseptic connectors typically are two-part constructions (either a male and matching female part or a pair of “genderless” parts) that are joined together. One part of the aseptic connect can be joined to the vessel, e.g., by a suitable sized length of tubing. This aseptic connector is then coupled to a corresponding aseptic connector part on the probe assembly, as described below. When the aseptic connector that is mounted on the container is connected to the aseptic connector on the sterilized probe sheath assembly, a sterile passageway is formed between the container and the probe sheath, a passageway through which a sterile sensor or probe can be inserted such that it can take measurements of conditions inside the vessel. 
     Plunger-Type Assembly 
     Turning now to  FIG.  1 A , a plunger-type probe assembly  100  is shown having a hollow probe sheath  20  which has a barbed tube fitting  24  at a first end  22  which is used to connect the probe sheath  20  to a disposable aseptic connector  26 , and the second end  28  of the probe sheath  20  provides an opening  32  into which a sensor or probe body  40  and plunger  34  is inserted. The disposable aseptic connector  26  is attached using a suitably sized section of tubing  36  to the barbed tube fitting  24  on the first end  22  of the probe sheath  20 , thereby forming a closure  38  at that distal end capable of maintaining a sterile seal. At the left, bag wall  64  has a port  72  formed in the exterior of the bag  512 . Port  72  may have a hose barb plate  70  welded to the inside of the bag wall  64 , and a valve protrusion, such as a hose barb  74 , projecting from the bag  512 . 
     The probe sheath plunger  34  has an opening  33  into which the sensor or probe body  40  can be inserted and secured such that a seal capable of being sterilized is formed between the sensor or probe body  40  and the probe sheath plunger  34 . The probe sheath plunger  34  is positioned inside the opening  32  of the probe sheath  20  such that the sensor/probe body  40  with the sensing element can pass through the inside  42  of the probe sheath  20  and reach to the barbed fitting  24  on the first end  22  of the probe sheath  20 . 
     As shown in  FIG.  1 B , a seal  44  exists between the inside  42  of the probe sheath  20  and the outside  35  of the probe sheath plunger  34  which allows relative movement between the probe sheath  20  and the probe sheath plunger  34 . The plunger  34  can be a manual plunger or a low-rpm, high-torque motor assembly  31 . The seal  44  between the probe sheath  20  and the probe sheath plunger  34  is configured so that the seal  44  will provide a sterile barrier between the volume  42  inside the probe sheath  20  and the outside of the probe sheath  20  when the probe sheath  20  is sterilized. 
     The probe sheath plunger  34  can be moved relative to the probe sheath  20  so that when the sensor or probe body  40  needs to be inserted through the wall  64  of a flexible or semi-rigid container, column, or tubing, the plunger  34  is moved such that it decreases the internal volume  42  inside the probe sheath  20 , and the sensor or probe body  40  then moves down the sheath to the disposable aseptic connector  26 . 
     As shown in  FIG.  1 B , the probe assembly  100  can further include a locking mechanism such as a threaded portion  48 , which portion  48  can include a catch, detent, for example, positioned to maintain the probe sheath plunger  34  in the fully compressed position, as shown in  FIG.  1 B , when the sensor or probe  40  is through the wall  64  of the flexible or semi-rigid container or tubing. Arrow  60  shows direction of movement of the elongate probe body  40  longitudinally in the direction of the bag  512 . 
     The bag  512  can have an entry point or port  72  formed in the exterior of the bag  512 . This port  72  can include a hose barb plate  70  welded to the inside of the bag wall  64  and a valve protrusion, such as hose barb  74  projecting from the bag  512 . The valve protrusion  74  may be integrally formed in the exterior of the bag  512 , for example by welding a hose barb  74  into the bag film  64  in a disposable bag type reactor. The valve protrusion  74  should releasably engage an aseptic connector  76 , such that the aseptic connector can mate with another portion of an aseptic connector  26 . The aseptic connector  76  can be connected in any suitable manner to protrusion  74 , so long as the connection does not leak.  FIG.  1 B  depicts a disposable aseptic connector part  76  mating with a hose barb  74  secured by a clamping mechanism  78 , such as a tri-clover type clamp. 
     The aseptic connector can include two separate portions, or parts  26 ,  76 . These portions can mate together in a traditional male and female relationship, as is shown in  FIG.  1 A . Other types of connectors may be used with the disclosed probe assembly. For example, the aseptic connector portions can connect to one-another in a non-mating fashion, such that each portion of the aseptic connector is identical. Clamping mechanisms can be utilized to ensure proper sealing and non-leaking function of the aseptic connector. The aseptic connector can include a non-permeable membrane sealing the connectors portions from contamination from the ambient environment, this membrane being designed to be removed prior to insertion of the probe body through the aseptic connector. The aseptic connector can be appropriately sized to match the diameter of a desired probe, vessel port, probe assembly connection size, or any other desired sizing variable. The type of aseptic connector can be selected without regard to the embodiment of the probe sheath type. Aseptic connectors are available from various commercial sources, such as Colder Products, Pall, Milllipore and GE Healthcare. 
     The probe sheath plunger  34  can be disposed within the probe sheath  20  such that no ambient air, liquids, or other matter from the exterior of sheath  20  can pass to the sheath interior  42 . The probe sheath plunger  34  can be formed of a rubber material such that the plunger can slide along the probe sheath  20  and such that the plunger  34  forms a seal directly against the probe sheath  20 . Alternatively, as explained above the assembly can include seals  44 . Alternatively, in another embodiment there is no plunger  34 ; instead, for example, a portion of the probe body serves as the actuator. In this case, the seals  44  contact the elongate probe body directly, aseptically sealing the interior  42  from the ambient environment. 
       FIG.  1 C  depicts another embodiment of the disclosed probe assembly wherein the probe sheath includes at least two parts, one of which comprises a tubular section of the probe sheath which can be removed after the probe sheath has been collapsed and the probe body is locked together with a bag port. 
     The embodiment in  FIG.  1 C  includes a plunger-type probe assembly  150  having a hollow probe sheath  20  having an inside wall, a first end  22 , and a second end  28  into which a rear plunger  34  has been inserted through opening  32 . Rear plunger  34  is secured to the inside wall of probe sheath  20 , for example by means of a bayonette fitting with prongs  62  (shown in  FIG.  1 D  but not shown in  FIG.  1 C ). The front portion of rear plunger  34  includes threads  35  arranged for mating and connecting to a threaded portion  48  of a connector plug  45  in the front portion of the probe sheath  20 . A sensor or probe body  40  is axially positioned within probe sheath  20  and secured at its rear end within rear plunger  34 , and secured at its front end within connector plug  45 . The front portion of sensor or probe body  40  is positioned in a disposable aseptic connector  26  which is attached, for example, using a suitably sized section of tubing to the barbed tube fitting  24  on the first end  22  of the probe sheath  20 , thereby forming a closure capable of maintaining a sterile seal. A detachable tool  509  to which is affixed handle  508  is connected to the probe sheath  20 . The seals  44  between the probe sheath  20  and the probe sheath plunger  34  are configured so that the seal  44  will provide a sterile barrier between the inside of the probe sheath  20  and the outside of the probe sheath  20  when the probe sheath  20  is sterilized. Seals  44  can be O-rings. 
     Arrow  624  shows the direction in which the rear plunger  34  is moved to advance the probe body  40  into the bag  512  as shown in  FIG.  1 D . 
       FIG.  1 D  shows the plunger type probe assembly  150  in a collapsed position from the position that is depicted in  FIG.  1 C . At the left, bag wall  64  of container  512  has a port  72  formed in the exterior of the bag  512 . Port  72  may have a hose barb plate welded to the inside of the bag wall  64 , and a valve protrusion, such as a hose barb  74 , projecting from the bag wall  64 . The sensor/probe body  40  with the sensing element has advanced, passing through the inside of the probe sheath  20 , and through the disposable aseptic connector  26  with end wall  38 , and through the bag wall  64  to the interior of bag  512 . Clamping mechanisms  78  secure the probe body  40 . 
       FIG.  1 D  also shows a connection port  36   a  of the disposable aseptic connector  26  connected to the barbed tube fitting  24  of the probe sheath  20 . The threaded portion  35  of probe sheath plunger  34  is shown mated with the threaded portion  48  of connector plug  45 . The tubular portion of probe sheath  20  has been removed, along with tool  509  with handle  508 , the tool having being used to detach the probe sheath  20 . The connector plug  45  is shown with the prongs  62  that had been used to attach the rear plunger  34  to the inside wall of probe sheath  20 . 
     Telescoping Assembly 
       FIG.  2 A  is a side elevation, partially cutaway view of a telescoping plunger probe assembly  200  according to another embodiment of the invention. The telescoping assembly  200  differs from the above description only in that the probe sheath  20  may consist of one or more sections  50  that allow the sections of the probe sheath  20  to be telescoped into one another, such that the overall length of the probe sheath  20  is reduced when the plunger  34  is fully compressed into the probe sheath  20 , as shown in  FIG.  2 B . Arrow  68  shows direction of movement of the elongate probe body  40  longitudinally in the direction of the bag wall  64 . The telescoping sections  50  are configured to provide moveable seals  52  between the sections  50 , and these seals  52  are such that they, along with the other seals  44  described above can provide a sterile interior space  42  formed inside the probe sheath  20  once the assembly has been sterilized. 
     As discussed above, the probe sheath assembly can again include a locking mechanism such as a threaded portion  48 , a catch, detent, etc., to maintain the telescoping sections  50  or segments of the probe sheath  20  in their fully compressed configuration as shown in  FIG.  2 B , or telescoped configuration when the probe sheath plunger  34  is in the fully compressed position and the sensor or probe is inserted into the flexible or semi-rigid container or tubing  64 . 
     Balloon Plunger-Type Assembly 
       FIGS.  3 A and  3 B  are side elevation, partially cutaway views of a balloon plunger-type probe assembly  500  according to another embodiment of the invention, wherein a rigid or semi-rigid sheath  502  is connected to a flexible sheath portion  504 . The flexible sheath portion  504  can be connected to an elongate sensor or probe body  506  or to an elongate probe handle  508  that is attached to the elongate probe body  506  disposed within the probe sheath  502 ,  504 . 
     The flexible portion  504  can be fixed to the rigid or semi-rigid sheath portion  502  with means known in the art, such as clamp  510 . The flexible portion  504  can be elastic, or inelastic, so long as it is deformable and is able to maintain a seal with sheath  502  when the probe body  506  or handle  508  is moved longitudinally in the direction of the bag  512 . When the probe  506  is disposed within the bag  512 , the flexible portion  504  can be disposed within the rigid or semi-rigid portion  502 , as shown in  FIG.  3 B . 
     The sensor used in a probe sheath  20  according to an embodiment of the invention can be any type of sensor. Non-limiting examples include conductivity, pH, dissolved oxygen, and turbidity sensors. 
     The probe sheath  20  according to an embodiment of the present invention facilitates the removal or retraction of a sensor from a flexible or semi-rigid container  64  or a flexible or semi-rigid tubing  64  so that the sensor can be sterilized and re-used in another device. 
     Probe Insertion Device Not Requiring A Gas-Permeable Membrane Connector 
     At the outset, this invention of the invention is described in its broadest overall aspects, with a more detailed description following. 
     A probe insertion device that does not require a gas permeable membrane connector is described. The probe insertion device  700   b ,  FIG.  5 B , according to an embodiment of the invention includes a thin-walled collapsible tubing  604   a ,  605   b  that can be folded in upon itself, as shown  FIGS.  5 B,  5 C, and  5 D . 
     One embodiment of a method of producing the disclosed probe insertion device is depicted in  FIGS.  4 A through  5 D .  FIG.  4 A  depicts a first starting component  600   a  including a disposable or non-disposable probe body  606  having an elongate probe handle  608  and which is partially inserted into a first end of a section of flexible tubing  604  and clamped in place by clamp  610   a . The section of autoclavable or irradiatable, weldable, thin-walled, flexible tubing  604  is of sufficient length to fit in a standard tubing welder. The second end of flexible tubing  604  is clamped by clamp  610   b  to a sterile, autoclavable, gas vent filter  612  having an open end partially inserted into the flexible tubing  604 . Filter  612  may be, for example, a 0.2 micron filter. 
       FIG.  4 B  depicts a second starting component  600   b  comprising a section of flexible tubing  605  having a plug  615  or welded closing at a first end. Flexible tubing  605 , in one embodiment of the invention, has a wall that is thicker than the wall of tubing  604 . The section of flexible tubing  605  is autoclavable or irradiatable and weldable and of sufficient length to fit in a standard tubing welder. Tubing  605  is clamped by clamp  610   c  or otherwise attached to a container tubing port  620  secured to wall  64  of flexible container  512  or to a tubing system. The flexible container  512  or tubing system has preferably been irradiated, autoclaved or otherwise sterilized. 
     The first and second starting components  600   a  and  600   b , respectively, are autoclaved, irradiated, or otherwise sterilized. The middle sections of the starting components are then placed in a standard tubing welder  625 .  FIG.  5 A  depicts an arrangement,  700   a , wherein a middle section of each of the first starting component  600 A and the second starting component  600   b  are positioned within a standard tubing welder  625 . Part i,  604   a  and Part ii,  605   b  are welded together at weld joint  622  as shown in  FIG.  5 B . The filter  612  of the first starting component  600   a  and the plugged tubing end of the second starting component  600   b  are discarded. 
     As shown in  FIG.  5 B , the probe insertion device  700   b  in  FIG.  5 B  has been formed from two flexible tubing parts, Part i and Part ii,  604   a  and  605   b , respectively, which have been autoclaved, irradiated, or otherwise sterilized along with a sacrificial gas permeable vent filter  612  that is discarded after the parts  604   a  and  605   b  are placed in a tube welder  625  and welded together. As also shown in  FIG.  4 B ,  FIGS.  5 A and  5 B , flexible tubing  605  and/or Part ii,  605   b  is clamped by clamp  610   c  or otherwise attached and fluidically connected to container tubing port  620  of flexible or semi-rigid container  512  or to a flexible tubing into which the probe  606  is to be inserted. After Parts i and ii,  604   a  and  605   b  respectively have been welded together as shown in  FIG.  5 B , the probe body  606  is advanced in the direction of arrow  624  and inserted into the container  512  by collapsing the flexible tubing  604 ,  604   a  inward as the probe is pushed or otherwise advanced through the weld, through tubing  605 ,  605   b  and through the container tubing port  620  and into the container  512 .  FIG.  5 C  shows the resulting probe assembly  700   c.    
       FIG.  5 C  shows that the flexible tubing Part i,  604   a  has folded back on itself, and is now positioned inside of flexible tubing Part ii,  605   b . Probe body  606  with attached probe wire  630  is shown as positioned or “sandwiched” between two surfaces formed of tubing  604   a . As shown in  FIG.  5 D , a perimeter or outer hose clamp  640  can be attached to secure the probe body  606  in position and to prevent backward leaking. 
     EQUIVALENTS 
     One skilled in the art will appreciate further features and advantages of the invention based on the above-described embodiments. Accordingly, the invention is not to be limited by what has been particularly shown and described, except as indicated by the appended claims. All publications and references cited herein are expressly incorporated herein by reference in their entirety.