In-SITU protein analysis device, system, and method

Abstract

An in-situ protein analysis device includes an inlet in fluid communication with a process pipeline of a bioreactor, the inlet being configured to receive samples from the process pipeline, a cartridge including a plurality of reservoirs, and a reaction system configured to create one or more analysis samples by selectively combining each sample with fluid from one or more of the plurality of reservoirs. The in-situ protein analysis device also includes an incubation system configured to receive the one or more analysis samples and a protein analyzer configured to receive and test the one or more analysis samples.

Description

IN-SITU PROTEIN ANALYSIS DEVICE, SYSTEM, AND METHODCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Patent Application No.63 / 553,265, filed February 14, 2024, which is hereby incorporated by reference in its entirety.BACKGROUND OF THE INVENTION

[0002] A variety of protein analysis systems and devices are commercially available. However, there is a need for improved protein analysis systems, devices, and methods.

[0003] The present invention provides for ameliorating at least some of the disadvantages of the prior art. These and other advantages of the present invention will be apparent from the description as set forth below.BRIEF SUMMARY OF THE INVENTION

[0004] In an aspect, the present disclosure provides an in-situ protein analysis device comprising an inlet in fluid communication with a process pipeline of a bioreactor, the inlet being configured to receive samples from the process pipeline, a cartridge including a plurality of reservoirs, and a reaction system configured to create one or more analysis samples by selectively combining each sample with fluid from one or more of the plurality of reservoirs. The in-situ protein analysis device also comprises an incubation system configured to receive the one or more analysis samples and a protein analyzer configured to receive and test the one or more analysis samples.

[0005] In an aspect, the present disclosure provides an in-situ protein analysis system comprising a bioreactor having a process pipeline, a connector configured to receive samples from the process pipeline of the bioreactor, and a cartridge including a plurality of reservoirs. The in-situ protein analysis system also comprises a reaction system configured to create one or more analysis samples by selectively combining each sample with fluid from one or more of theplurality of reservoirs, an incubation system configured to receive the one or more analysis samples, and a protein analyzer configured to receive and test the one or more analysis samples.

[0006] In an aspect, the present disclosure provides a method for in-situ protein analysis, the method comprising actuating one or more inlet valves in fluid communication with a process pipeline of a bioreactor to obtain samples from the process pipeline and actuating a reservoir valve of one or more valve assemblies, each valve assembly being in fluid communication with a reservoir containing an agent. The method also comprises mixing each sample with one or more agents received via actuating of the one or more valve assemblies to create an analysis sample, transferring each analysis sample to an incubator, and initiating protein analysis of each incubated analysis sample.BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS

[0007] Subject matter of the present disclosure will be described in even greater detail below based on the exemplary figures. All features described and / or illustrated herein can be used alone or combined in different combinations. The features and advantages of various implementations will become apparent by reading the following detailed description with reference to the attached drawings, which illustrate the following:

[0008] FIG. 1 schematically illustrates an in-situ protein analysis system according to an embodiment of the present disclosure;

[0009] FIG. 2 illustrates an in-situ protein analysis system with a first connector, an analysis device, and a cartridge according to an embodiment of the present disclosure;

[0010] FIG. 3 illustrates a tank with a lid-mounted in-situ protein analyzer according to an embodiment of the present disclosure;

[0011] FIG. 4 illustrates a tank with a pipe-mounted in-situ protein analyzer according to an embodiment of the present disclosure; and

[0012] FIG. 5 schematically illustrates an in-situ protein analysis system according to an embodiment of the present disclosure.DETAILED DESCRIPTION OF THE INVENTION

[0013] Conventional protein analysis systems and processes require equipment to be taken offline so that samples can be extracted and sent to an external laboratory, where samples are diluted to low concentration, and specimen concentration within the sample is subsequently measured. Conventional systems and processes further require using ultraviolet absorbance measurements, which are not capable of tracking low-to-high protein concentrations in tangential flow filtration. Furthermore, ultraviolet absorbance measurements may not be feasible for use in slurry samples. Off-line measurement also delays timely control of tangential flow filtration. Conventional protein analysis processes can also suffer from interference resulting from aggregated protein, which may become discolored, and which can lead to measurement errors. Embodiments of the present disclosure address deficiencies of conventional protein analysis systems and processes by providing improved protein analysis, which can be performed in-situ, thereby reducing or eliminating the time and resource burdens associated with conventional protein analysis.

[0014] A protein analyzer and protein analyzer system are provided that comprise separate functional units that can be assembled together. The separate functional units of the protein analyzer system can include a filtration unit, a controller and valve unit, a disposable cartridge, an incubation bay, and a spectrophotometer unit. The separate functional units of the protein analyzer system can allow for cost-effective protein analysis system assembly, maintenance / repair, and customization, as functional units may be manufactured, acquired, stored, and interchanged separately and as needed. For example, a protein analyzer that must be transitioned from a first type of protein analysis can have a disposable cartridge replaced with a new disposable cartridge that includes different reagents, thereby enabling the protein analyzer to conduct a different type of protein analysis without other expensive modification or timeintensive configuration. In another example, a malfunction in an incubation bay of a protein analyzer could be easily addressed by replacement of the malfunctioning incubation bay with a new incubation bay, rather than replacement of the entire protein analyzer, thereby reducing the time and cost burden of repair / replacement processes to bring protein analysis back online.

[0015] In an embodiment, a protein analysis device is provided which can be added to a preexisting bioreactor or fermentation tank to digitalize protein monitoring. Protein monitoringcan thus be accomplished by automatic measurements, and processing data can be acquired in real-time or near real-time (e.g., being delayed from real-time by the time required for electronic communication and automatic data processing). A protein assay can be integrated into a fully automatic fluidic system that includes a photometric or spectroscopic unit to achieve sample handling and measurement in a single closed-loop system without the need for human intervention.

[0016] In an embodiment, an enclosed system is provided that includes a disposable cartridge in which analytical reagents and a blank sample are carried. The enclosed system is a fluidic system that contains flow control valves, filters, valves (such as rotary valves), a mixing chamber, an incubation bay, a cooling bay, a photometric flow cell, a photometric or spectroscopic unit, and a fluidic pump. A sample is obtained directly from a source such as a bioreactor or fermentation tank, and is passed, e.g., pumped, into a location in which it is mixed with reagents at a controlled mixing ratio. The mixed sample is driven into an incubation bay and is maintained at an incubation temperature in the bay for an incubation period. The mixed sample is then passed into a cooling bay to return to room temperature. The mixed sample is subsequently brought into a photometric flow cell for photometric or spectroscopic detection and measurement. The foregoing features allow for a customized protein analysis to be carried out with minimal user involvement, thereby increasing testing availability and frequency, reducing the likelihood of human error intervening in an analysis process, and allowing users, who would otherwise be diverted from other duties in order to obtain samples for a remotely conducted protein analysis, to focus on other tasks.

[0017] A system according to an aspect of the present disclosure uses a protein assay, which is a method for colorimetric detection and quantitation of protein. In an illustrative application, the system is configured to detect specific protein molecules by using one or more specific biological assays. For example, a bicinchoninic acid (BCA) Protein Assay can be utilized, which combines a reduction of Cu2+ to Cu+ by protein in an alkaline medium with highly sensitive and selective colorimetric detection of Cu+ by BCA. One suitable example of a BCA protein assay is Pierce™, commercially available from ThermoFisher Scientific headquartered in Waltham, Massachusetts. A BCA assay is a copper-based colorimetric assay for total protein quantification. BCA assays rely on the formation of a Cu2+ protein complex in a basicenvironment, followed by reduction of the Cu2+ to Cu+. The amount of Cu2+ that is reduced is proportional to the amount of protein present in a tested solution. The response curve from a BCA assay allows accurate determination of unknown protein concentrations and provides a higher dynamic range than other standard assays.

[0018] In an embodiment, a protein analyzer is provided that is configured as an instrument that can be mounted to filtration systems, bioreactors, and / or fermenters. The protein analyzer can be mounted to previously installed, already operational, and / or existing filtration and bioreactor products and systems, allowing the protein analyzer to be installed without requiring complete replacement of preexisting hardware and thereby reducing installation time and costs. Filtration can be carried out, for example, as a pre-processing step or as an in-situ process via a centrifuge configured to remove large particles from a sample volume to avoid potential interference that may be caused by such particles. The protein analyzer is configured for integration with existing systems, such as processing chambers and / or processing pipes of the existing systems, via a connector that can be connected or retrofitted to the existing systems. The protein analyzer can provide for digitized in-situ protein analysis and can include embedded lab- on-a-chip systems for protein analysis. The protein analyzer can provide analysis results in time intervals, of, for example, a sampling rate of one measurement per 2-10 minutes per data point. The protein analyzer can provide for adoption of various assay kits including ligand-binding assays, immunoassays, and / or bioassays. As a result, real-time or near real-time measurements of concentrations of proteins, antigens, enzymes, and / or cells can be provided for.

[0019] FIG. 1 schematically illustrates an in-situ protein analysis system 100 according to an embodiment of the present disclosure. The system 100 includes a process pipeline 102 that is part of sub-system including a bioreactor and / or a fermenter. The process pipeline 102 can be any part of a network of devices, systems, and pipes that facilitate a biological reaction, and carries a fluid with biological material to be analyzed. A first connector 104 is provided that is connected or retrofitted to the process pipeline 102. The first connector 104 provides for fluid connection to a pre-existing process pipeline 102, thereby allowing the in-situ protein analyzer and analysis method to be implemented on various preexisting systems while reducing or entirely eliminating the need for modification to original system components or hardware. Specifically, the first connector 104 provides for a direct fluid connection to a process pipeline102 via which a sample may be directly obtained without interrupting fluid flow through the process pipeline 102 or disrupting fluid processing. Because the first connector 104 thereby enables in-situ protein analysis (as described in greater detail below), conventional protein analysis methods that require additional time and resource burdens associated with extracting and sending a sample to a remotely located facility or machine can be avoided. The first connector 104 provides for fluid connection to the process pipeline 102, thereby allowing a volume of fluid to be drawn from the process pipeline 102 as a sample. The first connector 104 can be a manifold or other similar form of connector. The first connector 104 can include standardized male or female pipe or tubing dimensions and shapes (e.g., threads or barbs) to facilitate compatibility with various off-the-shelf components, thereby reducing system cost and increasing ease of installation.

[0020] The sample volume is drawn from the process pipeline 102 via a pump 106 and passed to a tangential flow filtration (TFF) device comprising an inlet (feed inlet), a retentate outlet, and a permeate outlet, the device providing a retentate fluid flow path between the inlet and the retentate outlet, and a permeate flow path between the inlet and the permeate outlet, with a porous TFF membrane 108 across the permeate flow path. A variety of TFF devices and configurations are suitable and known in the art, such as hollow fiber modules, spiral-wound modules, and flat-plate modules (sometimes referred to a flat-plate cassettes).

[0021] The sample volume is passed parallel to the TFF membrane 108 and through the retentate outlet, and is further processed for analysis as described below. The permeate passing through the TFF membrane 108 can be returned to the process pipeline, and recirculated to the TFF device if desired. The TFF membrane 108 can be included as part of, for example, a Centramate™ Cassette, or the membrane 108 can be, for example, a Supor® poly ethersulfone (PES) membrane commercially available from Cytiva®, headquartered in Marlborough, Massachusetts. A filtration unit can include the TFF device (including the TFF membrane 108), the pump 106, and one or more tubes and / or connections for the pump 106 to draw the sample volume through the TFF device.

[0022] The system includes a plurality of shutoff valves 110 and a plurality of flow control valves 112 in order to provide for precise drawing of the sample volume, control of the sample volume though the system 100, and flow control of various fluids from reservoirs 114, 116, 118.For example, the sample volume for each measurement provided by the system can be between 50 microliters to 10 milliliters. The system can be operated at atmospheric pressure to reduce system complexity and thereby reduce system production and operational costs. The flow rate of the system can be controlled via the flow control valves 112 and shutoff valves 110 to be, for example, in the range from 100 microliters to 10 milliliters per minute. A first shutoff valve 110 is opened in order allow the sample volume to proceed through the system 100, and a flow control valve 112 regulates the flow rate of the sample volume.

[0023] The system includes at least one reagent reservoir 114, at least one buffer reservoir 116, and at least one blank reservoir 118, each of which is in fluid connection to a system line carrying the sample volume via a shutoff valve and a flow control valve. The individual shutoff and flow control valves of each reservoir 114, 116, 118 enables individual control of fluid flow from each reservoir 114, 116, 118, thereby enabling precise control of the amount of fluid from any given reservoir 114, 116, 118 being added to the system network and being communicated to the sample volume. The system further includes an air reservoir 120 that can be selectively connected to the system network via a shutoff valve. A mixer 122 mixes the sample volume and one or more fluids from reservoirs 114, 116, 118 and air from air reservoir 120, thereby forming a mixed sample volume that proceeds through the system to an incubation unit 124. The incubation unit 124 and mixer 122 can be integrated into a single block or structure, thereby facilitating a compact physical footprint of the system.

[0024] The incubation unit 124 is configured to receive the mixed sample volume from the mixer and incubate it in preparation for analysis via a spectrophotometer 132. The incubation unit 124 includes rotary valves 126 for diverting the mixed sample volume to one of a plurality of incubation coils 128, and also for merging incubated sample volumes from each of the plurality of incubation coils 128 to a single fluid connection. Incubated sample volumes are then passed through a cooling coil 130 before exiting the incubation unit 124. The rotary valves 126 can comprise, for example, Sakura E300 rotary valves commercially available from GMI Inc., headquartered at Ramsey, Minnesota, and / or SwitchEZ™ electric rotary valves, commercially available from Precigenome LLC, headquartered at San Jose, California. The rotary valves 126 can be built into and integrated with a manifold wherein channels and valves are embedded. The rotary valves 126 can be motor-driven high-performance liquid chromatography (HPLC) rotaryvalves. The incubation unit 124 can be configured as an incubation bay in which the heating and cooling coils are machined, and which is assembled together with one or more electrical heaters and cooling boards. The incubation unit 124 can achieve a high maximum temperature and a high temperature ramping rate in order to facilitate increased analysis efficiency and speed. For example, the incubation unit 124 can achieve a temperature range of between room temperature to up to 95 °C, with a temperature ramping rate of 2-20 °C per minute.

[0025] Incubated sample volumes are then passed to a spectrophotometer 132 for carrying out a protein analysis of the incubated sample volume. The spectrophotometer 132 includes a laser diode 134 configured to emit a light beam 136 toward a testing chamber 138 in which the incubated sample volume is arranged. The spectrophotometer 132 also includes a photodiode 142 for detecting a post-sample light beam 140. Once a sample is analyzed via the spectrophotometer 132, it is passed to a waste reservoir 148 by pressure provided via a pump 146, such as a peristatic pump, that is controlled by a flow speed controller 144.

[0026] It will be readily understood that FIG. 1 is a schematic illustration of the system 100 only, and that different components of the system can therefore be housed within the same housing or structure even if they are illustrated separately or far apart in FIG. 1 . For example, in some configurations, the waste reservoir 148 can be housed together with reservoirs 114, 116, 118, as will be illustrated below. The spectrophotometer 132 can comprise a modified benchtop spectrophotometer or a unit built with a laser diode, photo diode, and a flow cell. The spectrophotometer 132 can have pre-selected and fixed emission and absorption wavelengths, or can be scanned within a full spectrometer range. The spectrophotometer 132 can have a wavelength range of, e.g., from 280 nanometers to 285 nanometers, for detection of specific specimens. The spectrophotometer can have a power consumption rating of, e.g., from 10 to 250 Watts.

[0027] FIG. 2 illustrates an in-situ protein analysis system 200 with a second connector 202, an analysis device 204, and a cartridge 214 according to an embodiment of the present disclosure. The second connector 202 is connected to the analysis device 204 to provide a fluid connection of the analysis device to a processing pipeline of a bioreactor and / or fermenter. The second connector 202 can be embodied similar or identical in configuration to the first connector 104. The analysis device includes one or more control buttons 206 by which a user can controloperation of various functions of the analysis device. The analysis device 204 includes a power button 208 for controlling an on / off state of the analysis device 204 and / or for controlling various power states (such as a sleep or standby power states) of the analysis device 204. The analysis device 204 also includes a display 210 for displaying information to a user, which can be displayed via alphanumeric characters and / or graphics. The analysis device 204 includes a cartridge slot 212 into which a cartridge 214 can be inserted so that reservoirs in the cartridge can be put in fluid communication with internal fluidic channels of the analysis device 204.

[0028] The analysis device 204 can be formed generally as a box. For example, the analysis device 204 can be formed as a box with a width of, e.g., 15-25 inches38-64 cm), a height of, e.g., 6-12 inches (« 15-30 cm), and a depth of, e.g., 8-15 inches (~ 20-38 cm), thereby providing a compact form factor that can be readily fitted or retrofitted to a processing pipeline, a bioreactor, or a fermenter with optimal convenience to installation, maintenance, and / or process monitoring personnel. The analysis device 204 can have a lighter weight compared to conventional analysis devices, which are not optimized for weight. For example, the analysis device 204 can weigh under, e.g., 30 pounds (~ 14 kg), thereby providing a light-weight analysis device that minimizes the need for supporting structures and / or bulky fittings that would otherwise be necessary for securing heavier analysis equipment, while reducing stresses imposed on existing pipelines, tanks, and / or equipment.

[0029] The cartridge 214 includes a plurality of reservoirs that include at least a reagent reservoir 216 and a waste reservoir 218. In some embodiments, the cartridge 214 also includes a buffer reservoir and / or a blank reservoir. In an embodiment, the cartridge 214 is disposable and comprises a plastic injection molded structure. The cartridge 214 can be customized according to an application and / or end user’s analysis needs by providing varying types of reservoirs and varying quantities of reservoir liquid volumes, thereby providing a high degree of versatility for the in-situ protein analysis system 200 and making it compatible for diverse analysis requirements. The cartridge 214 includes between three and eight reservoirs, with each reservoir containing a different reagent and / or solution. Each reservoir can have an internal volume of, e.g., between 10-500 milliliters. The waste reservoir 218 can be larger than other reservoirs of the cartridge 214, thereby ensuring sufficient volume to receive waste produced after many individual measurements are made of multiple sample volumes.

[0030] The in-situ protein analysis system 200 is configured to automatically implement an entire BCA protein assay procedure, and provides many advantages over conventional protein analysis systems. The system 200 provides for hands-free and automatic operation, reducing time and resource outlays associated with conducting a protein analysis and with training users to conduct such analyses. The system 200 provides for self-calibration via inclusion of a blank reservoir in the cartridge 214, which enables blank reference measurements. Time and cost savings are achieved via multi-data measurements provided by the system 200 in real-time or near real-time. A disposable cartridge 214 can be provided for each processing batch, thereby allowing for repeated use and minimizing the amount of work between batches processed. The cartridges 214 are interchangeable and compliant with multiple assay selections, making the system 200 versatile and allowing targeted protein analysis for a variety of applications.

[0031] FIG. 3 illustrates an in-situ protein analysis system 300 having a tank 308 with a lidmounted in-situ protein analyzer 302 according to an embodiment of the present disclosure. The tank includes a lid 306 and is configured to carry a volume of bulk solution 310. The protein analyzer 302 is mounted onto the lid 306 and includes a sampling probe 304 that extends into the bulk solution 310. The protein analyzer 302, due to its in-situ configuration and being mounted directly to the tank 308, provides for real-time and / or near-real-time protein analysis of one or more samples drawn from the bulk solution 310 via the sampling probe 304.

[0032] FIG. 4 illustrates an in-situ protein analysis system 400 having a tank 408 with pipemounted protein analyzer 402 according to an embodiment of the present disclosure. The tank includes a lid 406 and is configured to carry a volume of bulk solution 410. The protein analyzer 402 is mounted to a pipe 404 connected to the tank 408. A sample can be drawn from the bulk solution via the pipe 404 and into the protein analyzer. The pipe 404 can be a dedicated sampling tube for providing a sample to the protein analyzer, or the pipe 404 can be part of a processing pipeline via which a sample is drawn, thereby ensuring that a representative sample from a process is drawn for analysis.

[0033] The in-situ protein analysis systems 300, 400 can be attached and detached from their mounting points without affecting the processing capabilities of the tank and / or system to which they are attached. The in-situ protein analysis systems 300, 400 can provide for a single batch of measurements per consumable cartridge or up to seven days of continuous measurements perconsumable cartridge. Data measurements can be obtained via the systems 300, 400 with a frequency of between 24 measurements per day up to 8,640 measurements per day.

[0034] FIG. 5 illustrates an in-situ protein analysis system 500 according to an embodiment of the present disclosure. The system 500 includes an inlet 502 via which a booster pump 504 draws a fluid. The fluid then flows into a pre-treatment cartridge. The system further includes a water purification cartridge 510 arranged downstream of the pre-treatment cartridge 506, and an ultrapure water cartridge 512 arranged downstream of the water purification cartridge 510. The water purification cartridge 510 can comprise, for example, a MicroPure™ cartridge commercially available from ThermoFisher Scientific. An outlet 518 for dispensing of fluid is arranged downstream of the ultrapure water cartridge 512. The system includes a plurality of third connectors 508 so that a sample volume can be drawn from a particular part of the process of the system 500. The third connectors 508 can be embodied as first connectors 104 and / or second connectors 202. The sample is drawn via one of the third connectors 508 into a protein analyzer 514. The protein analyzer 514 is configured to analyze a drawn sample using one or more reagents provided via a cartridge 516 inserted into the protein analyzer 514.

[0035] The disclosure is further illustrated by the following exemplary aspects. However, the disclosure is not limited by the following aspects.

[0036] (1) An in-situ protein analysis device comprising: an inlet in fluid communication with a process pipeline of a bioreactor, the inlet being configured to receive samples from the process pipeline; a cartridge including a plurality of reservoirs; a reaction system configured to create one or more analysis samples by selectively combining each sample with fluid from one or more of the plurality of reservoirs; an incubation system configured to receive the one or more analysis samples; and a protein analyzer configured to receive and test the one or more analysis samples.

[0037] (2) The device of aspect 1, wherein the plurality of reservoirs of the cartridge include at least a reagent reservoir, a buffer reservoir, and a blank sample reservoir.

[0038] (3) The device of aspects 1 or 2, wherein the reaction system includes a valve assembly in fluid communication with each of the plurality of reservoirs, each valve assembly including one or more reservoir valves that are actuatable to allow controlled injection of agents from the respective reservoir into each sample.

[0039] (4) The device of any one of aspects 1-3, wherein at least one of the plurality of reservoirs of the cartridge is a waste reservoir configured to receive the one or more analysis samples from the protein analyzer.

[0040] (5) The device of any one of aspects 1-4, wherein the protein analyzer is a spectrophotometer.

[0041] (6) The device of any one of aspects 1-5, wherein the reaction system includes a mixer, the mixer being configured to create the one or more analysis samples by mixing together each sample with fluid from one or more of the plurality of reservoirs.

[0042] (7) The device of any one of aspects 1-6, further comprising a connector fluidly connecting the inlet to the process pipeline.

[0043] (8) The device of any one of aspects 1-7, wherein the incubation system includes a plurality of incubating coils configured to heat the one or more analysis samples and at least one cooling coil configured to cool the one or more analysis samples to room temperature.

[0044] (9) An in-situ protein analysis system comprising: a bioreactor having a process pipeline; a connector configured to receive samples from the process pipeline of the bioreactor; a cartridge including a plurality of reservoirs; a reaction system configured to create one or more analysis samples by selectively combining each sample with fluid from one or more of the plurality of reservoirs; an incubation system configured to receive the one or more analysis samples; and a protein analyzer configured to receive and test the one or more analysis samples.

[0045] (10) The system of aspect 9, wherein the protein analyzer is configured to send a signal indicating a result of each test, and wherein the in-situ protein analysis system further comprises a processor configured to receive the signal to send the result to a display.

[0046] (11) The system of aspects 9 or 10, further comprising a probe arranged in the bioreactor and in fluid communication with the connector.

[0047] (12) The system of any of aspects 9-11, wherein the reaction system, the incubation system, and the protein analyzer are arranged within a single housing.

[0048] (13) The system of aspect 12, wherein the housing is mounted on the bioreactor.

[0049] (14) A method for in-situ protein analysis, the method comprising: actuating one or more inlet valves in fluid communication with a process pipeline of a bioreactor to obtain samples from the process pipeline; actuating a reservoir valve of one or more valve assemblies,each valve assembly being in fluid communication with a reservoir containing an agent; mixing each sample with one or more agents received via actuating of the one or more valve assemblies to create an analysis sample; transferring each analysis sample to an incubator; and initiating protein analysis of each incubated analysis sample.

[0050] (15) The method of aspect 14, further comprising inserting a cartridge including the one or more reservoirs into an in-situ protein analysis device.

[0051] (16) The method of aspect 15, further comprising actuating a waste valve in fluid communication with a waste reservoir, the waste reservoir being arranged in the inserted cartridge.

[0052] (17) The method of any of aspects 14-16, further comprising actuating a pump to draw the samples from the process pipeline through a filter.

[0053] (18) The method of any of aspects 14-17, further comprising heating each analysis sample in the incubator and cooling each sample to room temperature in the incubator.

[0054] (19) The method of any of aspects 14-18, wherein initiating the protein analysis of each incubated analysis sample comprises emitting a light beam into a testing chamber into which the analysis samples are received.

[0055] It shall be noted that the preceding aspects are illustrative and not limiting. Other exemplary combinations are apparent from the entirety of the description herein. It will also be understood by one of ordinary skill in the art that various embodiments may be used in various combinations with the other embodiments provided herein.

[0056] The use of the terms “a” and “an” and “the” and “at least one” and similar referents in the context of describing the invention (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. The use of the term “at least one” followed by a list of one or more items (for example, “at least one of A and B”) is to be construed to mean one item selected from the listed items (A or B) or any combination of two or more of the listed items (A and B), unless otherwise indicated herein or clearly contradicted by context. The terms “comprising,” “having,” “including,” and “containing” are to be construed as open-ended terms (i.e., meaning “including, but not limited to,”) unless otherwise noted. Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate valuefalling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”) provided herein, is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention.

[0057] Preferred embodiments of this invention are described herein, including the best mode known to the inventors for carrying out the invention. Variations of those preferred embodiments may become apparent to those of ordinary skill in the art upon reading the foregoing description. The inventors expect skilled artisans to employ such variations as appropriate, and the inventors intend for the invention to be practiced otherwise than as specifically described herein. Accordingly, this invention includes all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described elements in all possible variations thereof is encompassed by the invention unless otherwise indicated herein or otherwise clearly contradicted by context.

Claims

CLAIMS:

1. An in-situ protein analysis device comprising: an inlet in fluid communication with a process pipeline of a bioreactor, the inlet being configured to receive samples from the process pipeline; a cartridge including a plurality of reservoirs; a reaction system configured to create one or more analysis samples by selectively combining each sample with fluid from one or more of the plurality of reservoirs; an incubation system configured to receive the one or more analysis samples; and a protein analyzer configured to receive and test the one or more analysis samples.

2. The device of claim 1, wherein the plurality of reservoirs of the cartridge include at least a reagent reservoir, a buffer reservoir, and a blank sample reservoir.

3. The device of claims 1 or 2, wherein the reaction system includes a valve assembly in fluid communication with each of the plurality of reservoirs, each valve assembly including one or more reservoir valves that are actuatable to allow controlled injection of agents from the respective reservoir into each sample.

4. The device of any one of claims 1-3, wherein at least one of the plurality of reservoirs of the cartridge is a waste reservoir configured to receive the one or more analysis samples from the protein analyzer.

5. The device of any one of claims 1-4, wherein the protein analyzer is a spectrophotometer.

6. The device of any one of claims 1-5, wherein the reaction system includes a mixer, the mixer being configured to create the one or more analysis samples by mixing together each sample with fluid from one or more of the plurality of reservoirs.

7. The device of any one of claims 1-6, further comprising a connector fluidly connecting the inlet to the process pipeline.

8. The device of any one of claims 1-7, wherein the incubation system includes a plurality of incubating coils configured to heat the one or more analysis samples and at least one cooling coil configured to cool the one or more analysis samples to room temperature.

9. An in-situ protein analysis system comprising: a bioreactor having a process pipeline; a connector configured to receive samples from the process pipeline of the bioreactor; a cartridge including a plurality of reservoirs; a reaction system configured to create one or more analysis samples by selectively combining each sample with fluid from one or more of the plurality of reservoirs; an incubation system configured to receive the one or more analysis samples; and a protein analyzer configured to receive and test the one or more analysis samples.

10. The system of claim 9, wherein the protein analyzer is configured to send a signal indicating a result of each test, and wherein the in-situ protein analysis system further comprises a processor configured to receive the signal to send the result to a display.

11. The system of claims 9 or 10, further comprising a probe arranged in the bioreactor and in fluid communication with the connector.

12. The system of any of claims 9-11, wherein the reaction system, the incubation system, and the protein analyzer are arranged within a single housing.

13. The system of claim 12, wherein the housing is mounted on the bioreactor.

14. A method for in-situ protein analysis, the method comprising: actuating one or more inlet valves in fluid communication with a process pipeline of a bioreactor to obtain samples from the process pipeline; actuating a reservoir valve of one or more valve assemblies, each valve assembly being in fluid communication with a reservoir containing an agent; mixing each sample with one or more agents received via actuating of the one or more valve assemblies to create an analysis sample; transferring each analysis sample to an incubator; andinitiating protein analysis of each incubated analysis sample.

15. The method of claim 14, further comprising inserting a cartridge including the one or more reservoirs into an in-situ protein analysis device.

16. The method of claim 15, further comprising actuating a waste valve in fluid communication with a waste reservoir, the waste reservoir being arranged in the inserted cartridge.

17. The method of any of claims 14-16, further comprising actuating a pump to draw the samples from the process pipeline through a filter.

18. The method of any of claims 14-17, further comprising heating each analysis sample in the incubator and cooling each sample to room temperature in the incubator.

19. The method of any of claims 14-18, wherein initiating the protein analysis of each incubated analysis sample comprises emitting a light beam into a testing chamber into which the analysis samples are received.

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