bioreactor

Abstract

A bioreactor includes a vessel defining an interior chamber, a headplate configured to be detachably coupled to the vessel, an agitator coupled to the headplate and configured to extend into the interior chamber, at least one probe coupled to the headplate and configured to extend into the interior chamber, and at least one sparger coupled to the headplate and configured to extend into the interior chamber.

Description

BIOREACTORCROSS-REFERENCE TO RELATED APPLICATION(S)

[0001] The present application claims priority to and the benefit of U.S. Provisional Application No. 63 / 770,111, filed March 11, 2025, the entire content of which is incorporated herein by reference.BACKGROUND1. Field

[0002] The present disclosure relates to bioreactors.2. Description of the Related Art

[0003] Bioreactors are configured to grow organisms (such as yeast, bacteria, or animal cells) or other biochemically active substances in a controlled environment. Bioreactors may be utilized in industrial processes to produce pharmaceuticals (such as vaccines or antibodies) or to convert raw materials into useful byproducts, such as bio-converting com into ethanol. Related art bioreactors typically include a vessel, an agitator, and one or more sensor probes for measuring conditions inside the vessel. The organisms or other biochemically active substances growing in the vessel may be submerged in a liquid. Additionally, related art vessels are typically formed from stainless steel.

[0004] The above information disclosed in this Background section is only for enhancement of understanding of the background of the invention and therefore it may contain information that does not constitute prior art.SUMMARY

[0005] Aspects of the present disclosure relate to various embodiments of a bioreactor. In one embodiment, the bioreactor includes a vessel defining an interior chamber, a headplate configured to be detachably coupled to the vessel, an agitator coupled to the headplate and configured to extend into the interior chamber, at least one diptube coupled to the headplate and configured to extend into the interior chamber, and at least one sparger coupled to the headplate and configured to extend into the interior chamber.

[0006] The bioreactor may include a magnetic coupling assembly coupled to the headplate and the agitator.

[0007] The magnetic coupling assembly may include a support body, a lower magnet assembly in the support body and inside the interior chamber, and an upper magnet assembly above the lower magnet assembly and outside the interior chamber.-1- C:\Users\d_szakalski\ND Office EchoWAULT- E4JX9B16\ 197291 - 276370 (00003) - PCT Application -The lower magnet assembly is coupled to the agitator, the upper magnet assembly is configured to be coupled to a motor, and the lower magnet assembly is magnetically coupled to the upper magnet assembly.

[0008] The lower magnet assembly may include a lower casing and two or more lower magnets on the lower casing.

[0009] The upper magnet assembly may include an upper casing and two or more upper magnets on the upper casing.

[0010] The bioreactor may include a bayonet mount configured to detachably couple the support body to the headplate.

[0011] The bayonet mount may include radial pegs on the support body and corresponding L-shaped slots in the headplate.

[0012] The vessel comprises a polycarbonate material or a polyactic acid (PLA) material.

[0013] The agitator may include at least one impeller.

[0014] The impeller may be a pitched-blade impeller, a Rushton impeller, or a marine impeller.

[0015] The headplate may include a cap, a rim extending downward from a periphery of the cap, an annular groove in a lower surface of the rim, and threads on an inner surface of the rim.

[0016] The vessel may include a bottom wall, a sidewall extending upward from the bottom wall, a flange extending outward from the sidewall proximate to an upper end of the sidewall, an annular groove in an upper surface of the flange, and threads on an outer surface of the sidewall.

[0017] The sparger may be a micro-sparger or a macro-sparger.

[0018] The bioreactor may include ports in the headplate, and the probe may be coupled to the headplate at one of the ports.

[0019] At least one of ports in the headplate may be a threaded port.

[0020] The probe may include a threaded end configured to threadedly engage the threaded port in the headplate.

[0021] This summary is provided to introduce a selection of features and concepts of embodiments of the present disclosure that are further described below in the detailed description. This summary is not intended to identify key or essential features of the claimed subject matter, nor is it intended to be used in limiting the scope of the claimed subject matter. One or more of the described features may be combined with one or more other described features to provide a workable system or method.-2- C:\Users\d_szakalski\ND Office EchoWAULT- E4JX9B16\ 197291 - 276370 (00003) - PCT Application -BRIEF DESCRIPTION OF THE DRAWINGS

[0022] These and other features and advantages of embodiments of the present disclosure will become more apparent by reference to the following detailed description when considered in conjunction with the following drawings. In the drawings, like reference numerals are used throughout the figures to reference like features and components. The figures are not necessarily drawn to scale.

[0023] FIGS. 1A-1B are a top perspective view and a bottom perspective view, respectively, of a bioreactor according to one embodiment of the present disclosure;

[0024] FIGS. 2A-2B are perspective view and a cross-sectional view, respectively, of a vessel according to one embodiment of the present disclosure;

[0025] FIGS. 3A-3B are top perspective view and a bottom perspective view, respectively, of a headplate and probes according to one embodiment of the present disclosure;

[0026] FIGS. 4A-4B are top perspective view and a bottom perspective view, respectively, of a servo bracket according to one embodiment of the present disclosure;

[0027] FIGS. 5A-5B are top perspective view and a bottom perspective view, respectively, of a magnetic couple support according to one embodiment of the present disclosure;

[0028] FIG. 6 is a perspective view of an impeller according to one embodiment of the present disclosure;

[0029] FIGS. 7A-7D are a top perspective view, a bottom perspective view, an exploded perspective view, and a cross-sectional view, respectively, of a bioreactor according to another embodiment of the present disclosure;

[0030] FIGS. 8A-8B are top perspective view and a bottom perspective view, respectively, of a headplate and probes according to one embodiment of the present disclosure;

[0031] FIG. 80 is a bottom perspective view of a headplate according to one embodiment of the present disclosure; and

[0032] FIGS. 9A-9D are top perspective view, a bottom perspective view, a top exploded perspective view, and a bottom exploded perspective view, respectively, of a magnetic assembly according to one embodiment of the present disclosure.DETAILED DESCRIPTION

[0033] The present disclosure relates to various embodiments of a bioreactor. In one or more embodiments, the vessel of the bioreactor may include a polycarbonate / bioplastic material.-3- C:\Users\d_szakalski\ND Office EchoWAULT- E4JX9B16\ 197291 - 276370 (00003) - PCT Application -

[0034] Hereinafter, example embodiments will be described in more detail with reference to the accompanying drawings, in which like reference numbers refer to like elements throughout. The present invention, however, may be embodied in various different forms, and should not be construed as being limited to only the illustrated embodiments herein. Rather, these embodiments are provided as examples so that this disclosure will be thorough and complete, and will fully convey the aspects and features of the present invention to those skilled in the art. Accordingly, processes, elements, and techniques that are not necessary to those having ordinary skill in the art for a complete understanding of the aspects and features of the present invention may not be described. Unless otherwise noted, like reference numerals denote like elements throughout the attached drawings and the written description, and thus, descriptions thereof may not be repeated.

[0035] In the drawings, the relative sizes of elements, layers, and regions may be exaggerated and / or simplified for clarity. Spatially relative terms, such as “beneath,” “below,” “lower,” “under,” “above,” “upper,” and the like, may be used herein for ease of explanation to describe one element or feature’s relationship to another element(s) or feature(s) as illustrated in the figures. It will be understood that the spatially relative terms are intended to encompass different orientations of the device in use or in operation, in addition to the orientation depicted in the figures. For example, if the device in the figures is turned over, elements described as “below” or “beneath” or “under” other elements or features would then be oriented “above” the other elements or features. Thus, the example terms “below” and “under” can encompass both an orientation of above and below. The device may be otherwise oriented (e.g., rotated 90 degrees or at other orientations) and the spatially relative descriptors used herein should be interpreted accordingly.

[0036] It will be understood that, although the terms “first,” “second,” “third,” etc., may be used herein to describe various elements, components, regions, layers and / or sections, these elements, components, regions, layers and / or sections should not be limited by these terms. These terms are used to distinguish one element, component, region, layer or section from another element, component, region, layer or section. Thus, a first element, component, region, layer or section described below could be termed a second element, component, region, layer or section, without departing from the spirit and scope of the present invention.

[0037] It will be understood that when an element or layer is referred to as being “on,” “connected to,” or “coupled to” another element or layer, it can be directly on, connected to, or coupled to the other element or layer, or one or more intervening elements or layers may be present. In addition, it will also be understood that when an element or layer is referred to as being “between” two elements or layers, it can be the-4- C:\Users\d_szakalski\ND Office EchoWAULT- E4JX9B16\ 197291 - 276370 (00003) - PCT Application -only element or layer between the two elements or layers, or one or more intervening elements or layers may also be present.

[0038] The terminology used herein is for the purpose of describing particular embodiments and is not intended to be limiting of the present invention. As used herein, the singular forms “a” and “an” are intended to include the plural forms as well, unless the context clearly indicates otherwise. It will be further understood that the terms “comprises,” “comprising,” "includes," and "including," when used in this specification, specify the presence of the stated features, integers, steps, operations, elements, and / or components, but do not preclude the presence or addition of one or more other features, integers, steps, operations, elements, components, and / or groups thereof. As used herein, the term “and / or” includes any and all combinations of one or more of the associated listed items. Expressions such as “at least one of,” when preceding a list of elements, modify the entire list of elements and do not modify the individual elements of the list.

[0039] As used herein, the term "substantially," "about," and similar terms are used as terms of approximation and not as terms of degree, and are intended to account for the inherent variations in measured or calculated values that would be recognized by those of ordinary skill in the art. Further, the use of “may” when describing embodiments of the present invention refers to “one or more embodiments of the present invention.” As used herein, the terms "use," "using," and "used" may be considered synonymous with the terms "utilize," "utilizing," and "utilized," respectively. Also, the term “exemplary” is intended to refer to an example or illustration.

[0040] Unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention belongs. It will be further understood that terms, such as those defined in commonly used dictionaries, should be interpreted as having a meaning that is consistent with their meaning in the context of the relevant art and / or the present specification, and should not be interpreted in an idealized or overly formal sense, unless expressly so defined herein.

[0041] With reference now to FIGS. 1A-1B, bioreactor 100 according to one embodiment of the present disclosure includes a vessel 200 defining an interior chamber 201, a headplate 300 configured to be detachably coupled to an upper end of the vessel 200, an agitator 400 extending through the headplate 300 and into the interior chamber 201 of the vessel 200, a plurality of probes (sensors) 500 extending through the headplate 300 and into the interior chamber 201, one or more spargers 600 extending through the headplate 300 and into the interior chamber 201, a magnetic coupling assembly 700, and a micro-sparger assembly 800 inside the interior-5- C:\Users\d_szakalski\ND Office EchoWAULT- E4JX9B16\ 197291 - 276370 (00003) - PCT Application -chamber 201. In operation, the interior chamber 201 includes a liquid medium in which a bioactive process occurs in a controlled environment.

[0042] With reference now to the embodiment illustrated in FIGS. 2A-2B, the vessel 200 is a cylindrical body including a circular bottom wall 202 and a sidewall 203 extending upward from a periphery (e.g., an outer circumference) of the bottom wall 202. Together, the bottom wall 202 and the sidewall 203 define the interior chamber 201. In the illustrated embodiment, the vessel 200 also includes a flange (or lip) 204 extending outward from the sidewall 203 and around the periphery of the sidewall 203 proximate to the upper end of the sidewall 203, and threads 205 along an outer surface of the sidewall 203 above the flange 204. An upper surface of the flange 204 includes an annular groove or recess 206.

[0043] In one or more embodiments, the interior chamber 201 of the vessel 200 has a total volume in a range from approximately 2.5 liters (L) to approximately 4.7 L. In one embodiment, the interior chamber 201 of the vessel 200 may have a volume of approximately 3.6 L. Additionally, in one or more embodiments, the vessel 200 may have a working volume in a range from approximately 1.5 L to approximately 3.25 L.

[0044] In one or more embodiments, the vessel 200 may be formed from (or include) a polycarbonate / bioplastic material (e.g., a recycled polycarbonate material). In one or more embodiments, the vessel 200 may be formed from (or include) a polyactic acid (PLA) material or a PLA variant material.

[0045] With reference now to FIGS. 3A-3B, the headplate 300 according to one embodiment of the present disclosure includes a flat, circular cap 301 and a rim 302 extending downward from a periphery (e.g., a circumference) of the cap 301. A lower surface of the rim 302 of the headplate 300 includes an annular groove or recess 303. Additionally, an inner surface of the rim 302 includes threads 304. The threads 304 of the headplate 300 are configured to mate with the threads 205 of the vessel 200, and the annular groove 303 of the headplate 300 is configured to align (or substantially align) with the annular groove 206 of the vessel 200. Together, the annular groove 303 of the headplate 300 and the annular groove 206 of the vessel 200 are configured to accommodate a gasket 305 or other seal / sealant for forming a fluid-tight seal between the vessel 200 and the headplate 300.

[0046] The cap 301 of the headplate 300 also includes a plurality of ports 306 configured to accommodate the probes 500. The ports 306 may be threaded, nonthreaded openings, or a combination of threaded and non-threaded openings. Although in the illustrated embodiment the headplate 300 includes three ports 306, in one or more embodiments the headplate 300 may include any other suitable number of ports 306 depending, for instance, on the number of probes 500 (e.g., the headplate 300 may include from one to eight ports 306). The cap 301 of the headplate 300 also-6- C:\Users\d_szakalski\ND Office EchoWAULT- E4JX9B16\ 197291 - 276370 (00003) - PCT Application -includes one or more openings 307 configured to accommodate the one or more spargers 600. Although in the illustrated embodiment the headplate 300 includes four openings 307, in one or more embodiments the headplate 300 may include any other suitable number of openings 307 depending, for instance, on the number of spargers 600 (e.g., the headplate 300 may include from one to twelve openings 307).

[0047] In one or more embodiments, one of the probes 500 may be a pH probe configured to measure the acidity level within the interior chamber 201 of the vessel 200, and another one of the probes 500 may be a dissolved oxygen concentration (DO) probe configured to measure the DO within the interior chamber 201 of the vessel 200. Both pH and DO levels affect cell viability and metabolic activity and therefore the measurements taken by the pH probe and the DO probe may be utilized to ensure successful bioprocesses inside the vessel 200.

[0048] The one or more spargers 600 are configured to introduce gas (e.g., air, CO2, N2, and / or O2) into the interior chamber 201 of the vessel 200. Although in the illustrated embodiment the bioreactor 100 includes four spargers 600, in one or more embodiments the bioreactor 100 may include any other suitable number of spargers 600, such as from one to six spargers. The spargers may be micro-spargers, macrospargers, or a combination thereof depending, for instance, the type of bioreaction performed in the bioreactor 100. For instance, in one or more embodiments, one or more micro-spargers may be utilized for cell culture processes and one or more macrospargers may be utilized for fermentation processes.

[0049] With reference now to the embodiment illustrated in FIGS. 1A-1B, 4A-4B, and 5, the agitator 400 is an impeller-based agitator including a servo motor 401, a servo bracket 402 housing the servo motor 401, an axle 403 having an upper end coupled to the servo motor 401 and configured to be rotatably driven by the servo motor 401, and at least one impeller 404 coupled to a lower end of the axle 403. In one or more embodiments, the agitator 400 may include a first impeller 404(i) coupled to a lowermost end of the axle 403 and a second impeller 404(ii) coupled to the axle 403 above the first impeller 404(i). In one or more embodiments, the impeller 404 may include pitched impeller blades 405 (e.g., impeller blades pitched at approximately 45 degrees). In one or more embodiments, the impeller 404 may include any other suitable type or kind of impeller, such as a Rushton impeller (i.e., horizontal disk and flat, vertically-mounted blades on the horizontal disk) or a marine impeller.

[0050] In the illustrated embodiment, the headplate 300 also includes a support mount 308 extending upward from a central portion (e.g., the center) of the upper surface of the cap 301. The support mount 308 is configured to support and be coupled to the servo bracket 402. In the illustrated embodiment, the support mount 308 includes a generally rectangular standoff 309, a central opening 310, and four-7- C:\Users\d_szakalski\ND Office EchoWAULT- E4JX9B16\ 197291 - 276370 (00003) - PCT Application -mounting holes 311 in the corners of the standoff 309. The mounting holes 311 are configured to receive fasteners securing the servo bracket 402 to the support mount 308 of the headplate 300.

[0051] In the illustrated embodiment, the magnetic coupling assembly 700 includes a support body 701 and a magnet 702 (see FIG. 1B) supported on the support body 701. In the embodiment illustrated in FIGS. 5A-5B, the support body 701 includes a base 703, and annular sidewall 704 extending upward from the base 703, and a plurality of legs 705 extending outward from the annular sidewall 704. In one or more embodiments, the legs 705 may be regularly (i.e., equally or substantially equally) spaced apart from each other or irregularly spaced from each other. The legs 705 include openings 706 configured to accommodate fasteners coupling the magnetic coupling 700 to the headplate 300. The magnetic coupling 700 may include any suitable number of legs 705, such as from two to eight legs 705. When the magnetic coupling 700 is coupled to the headplate 300, the magnet 702 is inside the interior chamber 201 of the vessel 200. Additionally, in one or more embodiments, the bioreactor 100 may include a second magnet outside the interior chamber 201 and on top of the vessel 200.

[0052] In the illustrated embodiment, the micro-sparger 800 is inside the interior chamber 201 of the vessel 200 and it includes a plurality of suction cups 801 coupling the micro-sparger 800 to the bottom wall 202 of the vessel 200. The micro-sparger 800 is configured to introduce gas (e.g., air, CO2, N2, and / or O2) into the interior chamber 201 of the vessel 200.

[0053] In one or more embodiments, the bioreactor 100 may include an electrical thermal blanket wrapped around an outer periphery of the vessel 200.

[0054] In one or more embodiments, the vessel 200 of the bioreactor 100 may be formed by blow molding, such as extrusion blow molding, injection blow molding, or injection stretch blow molding.

[0055] In operation, the vessel 200 may be heated by an electrical thermal blanket. In one or more embodiments, the vessel 200 may be heated by the electrical thermal blanket to a maximum temperature of approximately 60 °C.

[0056] With reference now to FIGS. 7A-7D, a bioreactor 900 according to another embodiment of the present disclosure includes a vessel 1000 defining an interior chamber 1001, a headplate 1100 configured to be detachably coupled to an upper end of the vessel 1000, an agitator 1200 extending through the headplate 1100 and into the interior chamber 1001 of the vessel 1000, a plurality of diptubes 1300 extending through the headplate 1100 and into the interior chamber 1001, one or more spargers 1400 extending through the headplate 1100 and into the interior chamber 1001, and a magnetic coupling assembly 1500. In operation, the interior chamber 1001-8- C:\Users\d_szakalski\ND Office EchoWAULT- E4JX9B16\ 197291 - 276370 (00003) - PCT Application -includes a liquid medium in which a bioactive process occurs in a controlled environment. The vessel 1000 is shown transparent in FIGS. 7A-7B to reveal the agitator 1200, the diptubes 1300, and the sparger(s) 1400.

[0057] In the illustrated embodiment, the vessel 1000 is a cylindrical body including a circular bottom wall 1002 and a sidewall 1003 extending upward from a periphery (e.g., an outer circumference) of the bottom wall 1002. Together, the bottom wall 1002 and the sidewall 1003 define the interior chamber 1001. In the illustrated embodiment, the vessel 1000 also includes a flange (or lip) 1004 extending outward from the sidewall 1003 and around the periphery of the sidewall 1003 proximate to the upper end of the sidewall 1003, and threads 1005 along an outer surface of the sidewall 1003 above the flange 1004. An upper surface of the flange 1004 includes an annular groove or recess 1006.

[0058] In one or more embodiments, the sidewall 1003 of the vessel 1000 depicted in FIGS. 7A-7D may be taller than the sidewall of the vessel 200 depicted in FIGS. 1A-1 B. Additionally, in one or more embodiments, the total volume of the interior chamber 1001 of the vessel 1000 may be larger than total volume of the interior chamber 201 of the vessel 200 (e.g., the total volume may be greater than approximately 1.5 L, greater than approximately 2.5 L, greater than approximately 3.25 L, greater than approximately 3.6 L, or greater than approximately 4.7 L). The relatively taller sidewall 1003 (and consequently the relatively larger volume of the interior chamber 1001 ) may be configured to facilitate or accommodate a higher gas flow rate process, such as fermentation.

[0059] In one or more embodiments, the vessel 1000 may be formed from (or include) a polycarbonate / bioplastic material (e.g., a recycled polycarbonate material). In one or more embodiments, the vessel 1000 may be formed from (or include) a polyactic acid (PLA) material or a PLA variant material. In one or more embodiments, the polycarbonate or PLA material may be subjected to quality validation to ensure that the vessel 1000 will not degrade (or substantially not degrade) under the operating conditions of the bioreactor 900 and / or to prevent leaching of the material of the vessel 1000 into the interior chamber 1001 and the bioreactive material / solution contained therein (e.g., prevent cytotoxicity of the cells cultured in the bioreactor 900). In one or more embodiments, a lining (e.g., a spray in lining) may be included on an inner surface of the vessel 1000 to prevent (or at least mitigate) degradation of the vessel and / or leaching of the material of the vessel 1000 into the interior chamber 1001.Accordingly, unlike related art bioreactor vessels, which are commonly formed from metal (e.g., stainless steel) or glass to prevent degradation and leaching, the vessel 1000 according to one or more embodiments of the present disclosure may be formed-9- C:\Users\d_szakalski\ND Office EchoWAULT- E4JX9B16\ 197291 - 276370 (00003) - PCT Application -of (or include) a polycarbonate or PLA material, which are lighter, more affordable, and more readily recyclable compared to conventional materials.

[0060] With reference now to FIGS. 8A-8B, the headplate 1100 according to one embodiment of the present disclosure includes a flat, circular cap 1101 and a rim 1102 extending downward from a periphery (e.g., a circumference) of the cap 1101. A lower surface of the rim 1102 of the headplate 1100 includes an annular groove or recess 1103. Additionally, an inner surface of the rim 1102 includes threads 1104. The threads 1104 of the headplate 1100 are configured to mate with the threads 1005 of the vessel 1000, and the annular groove 1103 of the headplate 1100 is configured to align (or substantially align) with the annular groove 1006 of the vessel 1000. Together, the annular groove 1103 of the headplate 1100 and the annular groove 1006 of the vessel 1000 are configured to accommodate a gasket 1105 or other seal / sealant for forming a fluid-tight seal between the vessel 1000 and the headplate 1100.

[0061] The cap 1101 of the headplate 1100 also includes a plurality of ports 1106 configured to accommodate the diptubes 1300 and the one or more spargers 1400.The ports 1106 may be threaded, non-threaded openings, or a combination of threaded and non-threaded openings. In one or more embodiments in which the ports 1106 are threaded, an upper end portion of the diptubes 1300 and / or the sparger(s) 1400 may be threaded such that the diptubes 1300 and / or the sparger(s) 1400 may be threadedly coupled to the threaded ports 1106 in the headplate 1100. Although in the illustrated embodiment the headplate 1100 includes fourteen (14) ports 1106, in one or more embodiments the headplate 1100 may include any other suitable number of ports 1106 depending, for instance, on the number of diptubes 1300 and the number of spargers 1400 (e.g., the headplate 1100 may include from one to twenty ports 1106). In one or more embodiments, the number of ports 1106 may be greater than the total number of diptubes 1300 and spargers 1400 and the unused ports 1106 may be capped or plugged with threaded plugs 1107 (e.g., screwcaps) when not in use. In the illustrated embodiment, four threaded plugs 1107 are provided, but the present disclosure is not limited thereto. Although in the illustrated embodiment the headplate 300 includes one sparger 1400, in one or more embodiments the headplate 1100 may include any other suitable number of spargers 1400 depending, for instance, on the type of bioreactive process performed inside of the bioreactor 900 (e.g., the bioreactor 900 may include from one to four spargers 1400 ).The one or more spargers 1400 are configured to introduce gas (e.g., air, CO2, N2, and / or O2) into the interior chamber 1001 of the vessel 1000. The sparger(s) 1400 may be micro-sparger(s), macrospargers), or a combination thereof. The diptubes 1300 are configured to enable the introduction of media into the interior chamber 1001 of the bioreactor 900 and / or to-10- C:\Users\d_szakalski\ND Office EchoWAULT- E4JX9B16\ 197291 - 276370 (00003) - PCT Application -enable sampling the solution within the interior chamber 1001 by withdrawing a portion of the solution through one or more of the diptubes 1300.

[0062] The bioreactor 900 is also configured to support one or more probes (e.g., sensors) extending into the interior chamber 1001. The probes may be coupled (e.g., threadedly coupled) to the headplate 1101 by removing one or more of the threaded plugs 1107 and threading the one or more probes into one or more of the threaded ports 1106 in the headplate 1100. In one or more embodiments, the probes may be single use or multi-use (e.g., the probes may be multi-use probes that are sterilized between runs and then installed during the assembly and setup of the single-use vessel 1000). In one or more embodiments, one of the probes may be a pH probe configured to measure the acidity level within the interior chamber 1001 of the vessel 1000, and another one of the probes may be a dissolved oxygen concentration (DO) probe configured to measure the DO within the interior chamber 1001 of the vessel 1000. Both pH and DO levels affect cell viability and metabolic activity and therefore the measurements taken by the pH probe and the DO probe may be utilized to ensure successful bioprocesses inside the vessel 1000. Additionally, one or more of the probes may be a resistance temperature detector (RTD probe) configured to measure temperature by correlating it with increased electrical resistance in a metal element. In one or more embodiments, the RTD probe may be inserted into a tube 1301 having a capped bottom 1302 that serves as a thermowell.

[0063] With reference now to the embodiment illustrated in FIGS. 7A-8B, the agitator 1200 is an impeller-based agitator including a servo motor 1201, a servo bracket 1202 supporting the servo motor 1201, an axle 1203 having an upper end coupled to the servo motor 1201 and configured to be rotatably driven by the servo motor 1201 and a lower end coupled to the magnetic coupling assembly 1500, an impeller shaft 1204 having an upper end coupled to the magnetic coupling assembly 1500, and at least one impeller 1205 coupled to a lower end of the impeller shaft 1204.Although in the illustrated embodiment the agitator 1200 includes a single impeller 1205, in one or more embodiments, the agitator 1200 may include any other suitable number of impellers, such as two or more impellers. In one or more embodiments, the impeller 1205 may include pitched impeller blades 1206 (e.g., impeller blades pitched at approximately 45 degrees). In one or more embodiments, the impeller 1205 may include any other suitable type or kind of impeller, such as a Rushton impeller (i.e., horizontal disk and flat, vertically-mounted blades on the horizontal disk) or a marine impeller.

[0064] In the illustrated embodiment, the headplate 1100 also includes a support mount 1108 extending upward from a central portion (e.g., the center) of the upper surface of the cap 1101. The support mount 1108 is configured to support and be-11- C:\Users\d_szakalski\ND Office EchoWAULT- E4JX9B16\ 197291 - 276370 (00003) - PCT Application -coupled to the servo bracket 1202. In the illustrated embodiment, the support mount 1108 includes a generally rectangular standoff 1109, a central opening 1110, and two mounting openings 1111 in two of the comers of the standoff 1109. The mounting openings 1111 are configured to accommodate (receive) mounting pegs extending downward from the servo bracket 1202 to prevent rotation of the servo bracket 1202 relative to the headplate 1100. The servo bracket 1202 may have any configuration suitable for the type or kind of servo motor utilized.

[0065] With reference now to the embodiment illustrated in FIGS. 9A-9B, the magnetic coupling assembly 1500 includes a support body 1501, a lower magnet assembly 1502, and an upper magnet assembly 1503. The support body 1501 is in the interior chamber 1001 of the vessel 1000 and detachably coupled to the headplate 1100. The lower magnet assembly 1502 is supported on (above) the support body 1501 and is located in the interior chamber 1001 of the vessel 1000 between the support body 1501 and the headplate 1100. The upper magnet assembly 1503 is above the lower magnet assembly 1502 (e.g., spaced apart from the lower magnet assembly 1502) and is outside of the interior chamber 1001 of the vessel 1000. In one or more embodiments, the upper magnet assembly 1503 may be accommodated in the central opening 1110 of the headplate 1100. The upper magnet assembly 1503 is magnetically coupled to the lower magnet assembly 1502. In this manner, the magnetic coupling assembly 1500 couples the servo motor to the impeller shaft 1204 and the impeller 1205 such that, in operation, actuation of the servo motor rotates the impeller shaft 1204 and the impeller 1205.

[0066] In one or more embodiments, the support body 1501 may be detachably coupled to the headplate 1100 with a bayonet mount 1504. In one or more embodiments, the bayonet mount 1504 may include a pair of radial pegs 1505 (e.g., a pair of opposing radial pegs 1505) on the support body 1501 and a corresponding pair of L-shaped slots 1506 (e.g., a pair of opposing L-shaped slots 1506) in a cylindrical flange 1507 extending downward from the cap 1101 of the headplate 1100. The L-shaped slots 1506 in the cylindrical flange 1507 are configured to accommodate the pair of radial pegs 1505 on the support body 1501.

[0067] Additionally, in one or more embodiments, the magnetic coupling assembly 1500 also includes a radial bearing 1508. The radial bearing 1508 may be accommodated in a recess 1509 in the support body 1501. The impeller shaft 1204 extends up through an opening 1510 in the support body 1501, through a central opening 1511 of the radial bearing 1508, and through an opening 1512 in the lower magnet assembly 1502.

[0068] In one or more embodiments, the lower magnet assembly 1502 includes a lower casing 1513 and a plurality of lower magnets 1514 on the lower casing 1513.-12- C:\Users\d_szakalski\ND Office EchoWAULT- E4JX9B16\ 197291 - 276370 (00003) - PCT Application -The plurality of lower magnets 1514 may be arranged circumferentially around the lower casing 1513. Additionally, the plurality of lower magnets 1514 may be arranged with alternating polarities on the lower casing 1513. Similarly, in one or more embodiments, the upper magnet assembly 1503 includes an upper casing 1515 and a plurality of upper magnets 1516 on the upper casing 1515. The plurality of upper magnets 1516 may be arranged circumferentially around the upper casing 1515.Additionally, the plurality of upper magnets 1516 may be arranged with alternating polarities on the upper casing 1515.

[0069] In one or more embodiments, the bioreactor 900 may include an electrical thermal blanket wrapped around an outer periphery of the vessel 1000.

[0070] In one or more embodiments, the vessel 1000 of the bioreactor 900 may be formed by blow molding, such as extrusion blow molding, injection blow molding, or injection stretch blow molding.

[0071] In operation, the vessel 1000 may be heated by an electrical thermal blanket. In one or more embodiments, the vessel 1000 may be heated by the electrical thermal blanket to a maximum temperature of approximately 60 °C.

[0072] While this invention has been described in detail with particular references to exemplary embodiments thereof, the exemplary embodiments described herein are not intended to be exhaustive or to limit the scope of the invention to the exact forms disclosed. Persons skilled in the art and technology to which this invention pertains will appreciate that alterations and changes in the described structures and methods of assembly and operation can be practiced without meaningfully departing from the principles, spirit, and scope of this invention, as set forth in the following claims.-13- C:\Users\d_szakalski\ND Office EchoWAULT- E4JX9B16\ 197291 - 276370 (00003) - PCT Application -

Claims

WHAT IS CLAIMED IS:

1. A bioreactor comprising:a vessel defining an interior chamber;a headplate configured to be detachably coupled to the vessel;an agitator coupled to the headplate and configured to extend into the interior chamber;at least one diptube coupled to the headplate and configured to extend into the interior chamber; andat least one sparger coupled to the headplate and configured to extend into the interior chamber.

2. The bioreactor of claim 1 , further comprising a magnetic coupling assembly coupled to the headplate and the agitator.

3. The bioreactor of claim 2, wherein the magnetic coupling assembly comprises:a support body;a lower magnet assembly in the support body and inside the interior chamber; andan upper magnet assembly above the lower magnet assembly and outside the interior chamber,wherein the lower magnet assembly is coupled to the agitator,wherein the upper magnet assembly is configured to be coupled to a motor, andwherein the lower magnet assembly is magnetically coupled to the upper magnet assembly.

4. The bioreactor of claim 3, wherein the lower magnet assembly comprises a lower casing and a plurality of lower magnets on the lower casing.

5. The bioreactor of claim 4, wherein the upper magnet assembly comprises an upper casing and a plurality of upper magnets on the upper casing.

6. The bioreactor of claim 3, further comprising a bayonet mount configured to detachably couple the support body to the headplate.

7. The bioreactor of claim 6, wherein the bayonet mount comprises radial pegs on the support body and corresponding L-shaped slots in the headplate.-14- C:\Users\d_szakalski\ND Office EchoWAULT- E4JX9B16\ 197291 - 276370 (00003) - PCT Application -8. The bioreactor of claim 1 , wherein the vessel comprises a polycarbonate material.

9. The bioreactor of claim 1 , wherein the vessel comprises a polyactic acid (PLA) material.

10. The bioreactor of claim 1 , wherein the agitator comprises at least one impeller.

11. The bioreactor of claim 10, wherein the at least one impeller is a pitched-blade impeller, a Rushton impeller, or a marine impeller.

12. The bioreactor of claim 1 , wherein the headplate comprises:a cap;a rim extending downward from a periphery of the cap;an annular groove in a lower surface of the rim; andthreads on an inner surface of the rim.

13. The bioreactor of claim 12, wherein the vessel comprises:a bottom wall;a sidewall extending upward from the bottom wall;a flange extending outward from the sidewall proximate to an upper end of the sidewall;an annular groove in an upper surface of the flange; andthreads on an outer surface of the sidewall.

13. The bioreactor of claim 1 , wherein the sparger comprises a micro-sparger.

14. The bioreactor of claim 1 , wherein the sparger comprises a macro-sparger.

15. The bioreactor of claim 1 , further comprising a plurality of ports in the headplate, wherein the plurality of ports are configured to couple a plurality of probes to the headplate.

17. The bioreactor of claim 15, wherein at least one of the plurality of ports in the headplate is a threaded port.-15- C:\Users\d_szakalski\ND Office EchoWAULT- E4JX9B16\ 197291 - 276370 (00003) - PCT Application -18. The bioreactor of claim 17, wherein the at least one probe comprises a threaded end configured to threadedly engage the threaded port in the headplate.-16- C:\Users\d_szakalski\ND Office EchoWAULT- E4JX9B16\ 197291 - 276370 (00003) - PCT Application -

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