Chromatography apparatus and methods

Abstract

The present invention relates to a computer-implemented method 600 performed by a chromatographic device 400 configured to separate molecules of varying sizes from an eluent, the method comprising: obtaining (610) reference data for a chromatographic column of the chromatographic device, wherein the reference data indicates elution characteristics for a set of molecular sizes; obtaining (620) an elution progress measure and a corresponding quantitative measure indicating the concentration of molecules in an eluate of the chromatographic device; estimating (630) a size measure indicating the molecular size of the molecules in the eluate; rendering (640) a representation (300) indicating the quantitative measure and the size measure; and controlling (650) a display to display the representation to a user of the chromatographic device.

Description

Technical Field

[0001] This invention relates to a method for controlling a chromatographic apparatus. The invention further relates to a chromatographic apparatus for performing this method. Background Technology

[0002] Chromatography is a well-known process for analyzing and preparing chemical mixtures or samples. The sample is typically suspended in a fluid called a buffer composition or resin. The various sample components of the mixture travel down the chromatographic column at different speeds, thus separating them. This process is sometimes called size exclusion chromatography.

[0003] In size exclusion chromatography, macromolecules elute first because they have no access to the pores in the resin and therefore have access to smaller volumes. Small molecules diffuse into the pores and therefore have access to larger volumes as they pass through the column. This is further explained, for example, in https: / / en.wikipedia.org / wiki / Size-exclusion_chromatography.

[0004] This separation can be used to separate sample components in a fractionation step, where the mobile phase, known as elution, can be directed to one or more different fractionation vessels, for example, via the outlet valve of the chromatographic apparatus.

[0005] The problem with conventional solutions is that, in order to estimate the molecular weight of molecules in different containers, users are forced to perform post-elution and post-fractionation analyses, where the chromatographic process time or eluent volume is analyzed to estimate the molecular weight of molecules in different containers. Performing such post-elution analyses is a cumbersome and time-consuming operation.

[0006] Another problem with conventional solutions is that it is not possible to activate or deactivate fractionation based on the estimated molecular weight of the molecule to collect or discard the molecule separately.

[0007] Therefore, there is a need for an improved method for controlling fractionation in chromatographic equipment.

[0008] Purpose of the invention

[0009] The purpose of the embodiments of the present invention is to provide a solution that alleviates or solves the above-mentioned disadvantages and problems. Summary of the Invention

[0010] The above and other objectives are achieved through the subject matter described herein. Further advantageous embodiments of the invention are further defined herein.

[0011] According to a first aspect of the invention, the above and other objectives are achieved by a computer-implemented method executed by a chromatographic apparatus configured to separate molecules of varying sizes from an eluent, the method comprising: obtaining reference data for a chromatographic column of the chromatographic apparatus, wherein the reference data indicates elution characteristics for a set of molecular sizes; obtaining a elution progress measure and a corresponding quantitative measure indicating the concentration of molecules in the eluent of the chromatographic apparatus; estimating a size measure indicating the molecular size of molecules in the eluent; rendering a representation indicating the quantitative and size measures; and controlling a display to show the representation to a user of the chromatographic apparatus.

[0012] In a first embodiment according to the first aspect, molecular size is estimated using predetermined relationships and elution progress measures included in the reference data.

[0013] In the second embodiment according to the first embodiment, the predetermined relationship indicator curve and the predetermined relationship are obtained by performing curve fitting on a pair of values ​​including the gel phase distribution coefficient and the corresponding molecular size.

[0014] In the third embodiment according to the second embodiment, the elution progress measure is the elution volume, wherein the gel phase distribution coefficient depends on the total volume of the column, the elution volume, and the column void volume.

[0015] In the fourth embodiment according to the third embodiment, the gel phase distribution coefficient Kav is given by the relationship Kav=((Ve-V0)) / ((Vt-V0)), where Ve is the elution volume, V0 is the column void volume, and Ve is the elution volume.

[0016] In a fifth embodiment of any of the first to fourth embodiments, the predetermined relation is a polynomial relation.

[0017] In a sixth embodiment according to the first aspect or any of the foregoing embodiments, a quantitative measurement indicates the ultraviolet light absorption of the eluent.

[0018] In the seventh embodiment according to the first aspect or any of the foregoing embodiments, the elution progress measurement indicates the elution process run time or the volume of eluted liquid.

[0019] In the eighth embodiment according to the first aspect or any of the foregoing embodiments, the representation is a simplified diagram having quantitative measures represented on a first axis and an estimate of molecular weight on a second axis.

[0020] In a ninth embodiment of any of the embodiments according to the first aspect or the foregoing embodiments, the method further includes using molecular size and one or more predetermined intervals of molecular size to activate or deactivate fractionation.

[0021] At least one advantage of the first aspect is that the molecular weight and / or molecular size of molecules in the eluent can be dynamically determined, thereby avoiding the tedious, complex, and time-consuming operation of post-fractionation analysis. Another advantage is that the chromatographic process can be shortened by dynamically rendering and presenting a representation of molecular weight and / or molecular size to the user in real time, allowing the user to terminate the chromatographic process once the eluent containing the expected molecular weight and / or molecular size has been obtained again.

[0022] According to a second aspect of the invention, the above and other objectives are achieved by a chromatographic apparatus configured to separate molecules of varying sizes from an eluent, the apparatus comprising a column, a display, and a control unit including a circuit system. The circuit system includes a processing circuit system and a memory containing instructions executable by the processing circuit system, thereby configuring the chromatographic apparatus to perform the method according to the first aspect.

[0023] The advantages of the second aspect of the invention are at least the same as those of the first aspect of the invention.

[0024] The following detailed description of one or more embodiments will provide those skilled in the art with a more comprehensive understanding of embodiments of the invention and their additional advantages. It should be understood that similar reference numerals are used to identify one or more similar elements illustrated in the drawings. Attached Figure Description

[0025] Figure 1 Examples of simplified diagrams rendered and / or displayed by existing chromatographic equipment are shown.

[0026] Figure 2A -B shows an example of a standard simplified diagram.

[0027] Figure 3 A simplified diagram is shown according to one or more embodiments of this disclosure.

[0028] Figure 4 A chromatographic apparatus according to one or more embodiments of the present disclosure is shown.

[0029] Figure 5 A control unit according to one or more embodiments of the present invention is shown.

[0030] Figure 6 A flowchart illustrating a method according to one or more embodiments of the present disclosure is shown.

[0031] Figure 7 Predetermined relationships are shown according to one or more embodiments of this disclosure.

[0032] The following detailed description of one or more embodiments will provide those skilled in the art with a more comprehensive understanding of embodiments of the invention and their additional advantages. It should be understood that similar reference numerals are used to identify one or more similar elements illustrated in the drawings. Detailed Implementation

[0033] The word "or" in this description and corresponding claims should be understood to encompass both "and" and "or" in mathematical OR, rather than to be understood as XOR (different OR). The indefinite article "a" in this disclosure and claims is not limited to "a," but can also be understood as "one or more," i.e., multiple.

[0034] In this disclosure, references to containers or reservoirs will be used interchangeably, referring to receivers suitable for holding fluids.

[0035] In this disclosure, processing circuit systems and processing components are interchangeably referenced.

[0036] In this disclosure, the term "size measure" is a measure that indicates molecular volume and / or molecular weight, such as the molecular volume and / or molecular weight of molecules included in the eluent and processed during chromatography. Examples of such size measures are estimated molecular weight and / or estimated molecular size.

[0037] Figure 1 A simplified diagram rendered and / or displayed by a prior art chromatography apparatus 400 is shown. This diagram includes a vertical axis 110 indicating a quantitative measure, which indicates the concentration of molecules in the eluent. For example, the measure of UV light absorption of the eluent appearing from the chromatographic column of the chromatography apparatus 400. In other words, the quantitative measure indicates the concentration of molecules included in the eluent, which is recorded by a sensor (e.g., a sensor that measures absorbed light (e.g., UV light)). The diagram further includes a horizontal axis 120 indicating a measure of the progress of the chromatographic and / or fractionation process. Examples of progress measures could be chromatographic process time or eluent volume. (The last sentence appears to be incomplete and possibly refers to a different diagram.) Figure 1 We can see that three peaks, 131, 132, and 133, can be recorded for quantitative measurement. Each peak typically corresponds to a specific molecule with a specific molecular weight and size.

[0038] However, even if the quantitative measure is visualized / displayed / presented to the user of the chromatographic device 400, the user lacks the means to assess the molecular weight and / or molecular size of any molecule represented by peaks 131, 132, and 133, and is required to rely on process time or eluent volume. In other words, the user may, for example, manually control fractionation based on peaks 131, 132, and 133, but will have to rely on post-fractionation analysis to identify the molecular weight and / or molecular size of any molecule represented by peaks 131, 132, and 133.

[0039] In one example, a user is interested in using size exclusion chromatography to extract proteins with a specific molecular weight. The user will then have to perform a chromatographic and fractionation process or a chromatographic run, and then perform a post-fractionation analysis to identify the desired protein. This post-fractionation analysis is typically performed by obtaining a calibration curve and applying a formula to convert the chromatographic process time or eluent volume into molecular weight and / or molecular size.

[0040] Performing post-fractionation analysis is a tedious, complex, and time-consuming operation, and further requires that the chromatographic process be completed before the molecular weight and / or molecular size of the molecules in the eluent can be determined.

[0041] Figure 2A An example of a standard simplified diagram is shown. (As shown relative to...) Figure 1 The diagram has a vertical axis 110 and a horizontal axis 120. Furthermore, the diagram includes a display box detailing the quantitative measurement value Y (vertical axis) and the progress measurement value X (horizontal axis) marked 210 on the displayed curve 220. The curve indicates the quantitative measurement value and its corresponding progress measurement. In this example, the quantitative measurement Y is given in absorbance units mA. The progress measurement value X is given in chromatographic eluent volume Ve.

[0042] Figure 2B An example of a standard simplified diagram is shown. (As shown relative to...) Figure 1 The diagram has a vertical axis 110 and a horizontal axis 120. Furthermore, the diagram includes a display box detailing the quantitative measurement value Y (vertical axis) and the progress measurement value X (horizontal axis) marked 210 on the displayed curve 220. The curve indicates the quantitative measurement value and its corresponding progress measurement. In this example, the quantitative measurement Y is given in absorbance units mA. The progress measurement value X is given in chromatographic process time Te.

[0043] Figure 3 A simplified diagram 300 is shown according to one or more embodiments of the present disclosure. In the methods of the present disclosure, reference data indicating elution characteristics for a set of molecular sizes are used to dynamically determine size measures, such as estimating molecular weight and / or estimating molecular size, using the chromatographic process time or eluent volume of the chromatographic apparatus 400.

[0044] The representation is then rendered and / or displayed. This representation typically indicates quantitative and / or dimensional measurements in the diagram 300.

[0045] Schematic diagram 300 includes a vertical axis 310 indicating a quantitative measure that indicates the concentration of molecules in the eluent. For example, relative to... Figure 1 The vertical axis shown in Figure 2 is similar. This simplified diagram further includes a horizontal axis 320, which indicates the magnitude measurement. In this example, it is expressed in atomic mass units of Daltons or kilodaltons (kDa).

[0046] This has the following advantages: by dynamically providing users with information about the size and / or molecular weight of the molecules included in the eluent, it accelerates the size exclusion chromatography process and makes the process more efficient.

[0047] Figure 4 A chromatographic apparatus 400 according to one or more embodiments of the present disclosure is illustrated. The chromatographic apparatus 400 typically includes at least one inlet 455. The inlet may optionally be coupled to a reservoir 451 configured to hold a fluid (e.g., eluent). The inlet 455 may be implemented, for example, as a tubular element (e.g., a tube or hose). The inlet 455 may be configured to be coupled to a column 441. The column 441 may be included within the chromatographic apparatus 400 or disposed outside the chromatographic apparatus 400.

[0048] The chromatography apparatus 400 may further include a control unit 410, which includes a circuit system (e.g., a processor) and a memory. The memory may contain instructions executable by the processing circuit system, thereby operating the control unit 410 and / or the chromatography apparatus to perform any step or method described herein. Relative to Figure 5 The control unit 410 is described further.

[0049] Chromatographic apparatus 400 may optionally include a separator 470 coupled to the fluid outlet of column 441 and optionally to UV sensor 431, conductivity sensor 432, and outlet valve 420. Separator 470 may be configured to direct fluid received from column 441 to any of UV sensor 431, conductivity sensor 432, and outlet valve 420. Optionally, separator 470 may be communicatively coupled to a control unit and perform fluid coupling to UV sensor 431, conductivity sensor 432, and outlet valve 420 in response to one or more control signals from control unit 410.

[0050] UV sensor 431 may be communicatively coupled to control unit 410 and configured to measure quantitative quantities (e.g., UV light absorbance of the fluid) provided by separator 470. Chromatographic apparatus 400 may further include conductivity sensor 432, which is communicatively coupled to control unit 410 and configured to measure quantitative quantities (e.g., conductivity of the fluid) provided by separator 470. UV sensor 431 and / or conductivity sensor 432 may be further configured to provide the measured quantitative quantity to control unit 410 as a control signal including measurement data.

[0051] The chromatographic apparatus 400 may further include an outlet valve 420 coupled to the separator 470. The outlet valve 420 may have one or more outlets or outlet ports 421-423 and is configured to supply fluid supplied by the separator 470 to one or more outlets 421-423 in response to a control signal received, for example, from the control unit 410. In other words, fractionation of the eluent is performed.

[0052] Figure 5 A control unit 410 according to one or more embodiments of the present invention is illustrated. The control unit 410 may take the form of, for example, an electronic control unit, a server, an onboard computer, a fixed computing device, a laptop computer, a tablet computer, a handheld computer, a wrist-worn computer, a smartwatch, a smartphone, or a smart TV. The control unit 410 may include a processing circuitry 412 communicatively coupled to a transceiver 404 configured for wired or wireless communication. The control unit 410 may further include at least one optional antenna (not shown). The antenna may be coupled to the transceiver 404 and configured to transmit and / or transmit and / or receive wired or wireless signals in a communication network (e.g., WiFi, Bluetooth, 3G, 4G, 5G, etc.). In one example, the processing circuitry 412 may be any of the following selections: a processing circuitry system and / or a central processing unit and / or a processor module and / or multiple processors configured to cooperate with each other. Furthermore, the control unit 410 may further include a memory 415. The memory 415 may include, for example, hard RAM, a disk drive, a floppy disk drive, a flash drive, or other removable or fixed media drives, or any other suitable memory known in the art. Memory 415 may contain instructions executable by the processing circuitry system to perform any step or method described herein. Processing circuitry system 412 may be communicatively coupled to any of the transceiver 404, memory 415, pH sensor 431, conductivity sensor 432, outlet valve 420, and separator 470. Control unit 410 may be configured to send / receive control signals directly to any of the aforementioned units or to an external node, or via wired and / or wireless communication networks.

[0053] The wired / wireless transceiver 404 and / or the wired / wireless communication network adapter can be configured to send / receive data values ​​or parameters as signals to and from the processing circuitry system 412 and to other external nodes. For example, measured quantitative measurements.

[0054] In this embodiment, transceiver 404 communicates with external nodes directly or via a wireless communication network.

[0055] In one or more embodiments, the control unit 410 may further include an input device 417 configured to receive input or instructions from a user and to send a user input signal instructing the user to the processing circuitry system 412.

[0056] In one or more embodiments, the control unit 410 may further include a display 418 configured to receive a display signal from the processing circuitry 412 indicating a rendering object (e.g., a text or graphical user input object) and to display the received signal as an object (e.g., a text or graphical user input object).

[0057] In one embodiment, the display 418 is integrated with the user input device 417 and configured to receive a display signal from the processing circuitry 412 indicating a rendered object (e.g., a text or graphical user input object), and to display the received signal as an object (e.g., a text or graphical user input object), and / or configured to receive input or indication from the user, and to send a user input signal indicating user input or indication to the processing circuitry 412.

[0058] In another embodiment, the control unit 410 may further include and / or be coupled to one or more additional sensors (not shown), the one or more additional sensors being configured to receive and / or obtain and / or measure physical properties related to the chromatography apparatus 400, and to send one or more sensor signals indicative of the physical properties to the processing circuitry system 412. For example, a temperature sensor that measures ambient air temperature.

[0059] In one or more embodiments, the processing circuitry 412 is further communicatively coupled to the input device 417 and / or the display 418 and / or additional sensors and / or relative to... Figure 4 Any unit of the unit.

[0060] In one embodiment, a chromatographic apparatus 400 is provided and configured to separate molecules of varying sizes from an eluent. The chromatographic apparatus 400 includes a column 441, a display 418, and a control unit 410 including a circuitry system 412 and a memory 415. The memory 415 includes or contains instructions executable by the processing circuitry system 412, thereby configuring the chromatographic apparatus 400 to perform any of the method steps described herein.

[0061] Figure 6 A flowchart illustrating a method according to one or more embodiments of the present disclosure is shown. A computer-implemented method 600 is performed by a chromatographic apparatus 400 configured to separate molecules of varying sizes from an eluent. The method includes:

[0062] Step 610: Obtain reference data for the chromatographic column of the chromatographic apparatus, wherein the reference data indicates the elution characteristics for a set of molecules and the corresponding molecular sizes. Elution characteristics for a set of molecular sizes may include, for example, the aggregation values ​​for a specific molecular size and the chromatographic process progress time or run time from when the sample is introduced into column 411 until the expected molecules of a specific molecular size appear in the eluent and / or leave the column of chromatographic apparatus 400. Elution characteristics for a set of molecular sizes may further include, for example, the chromatographic process progress volume (e.g., eluent volume) measured from when the sample is introduced into column 411 until the expected molecules of a specific molecular size appear in the eluent and / or leave the column of chromatographic apparatus 400.

[0063] In one embodiment, reference data is obtained by retrieving reference data from memory (e.g., the memory of control unit 415). In another embodiment, reference data is obtained by receiving control signals from another node, such as a server, virtual server, or cloud server. Control signals may be sent / received, for example, via a communication network.

[0064] In one embodiment, the reference data includes any selected polymer, such as molecular identity, molecular weight, molecular size, volume, chromatographic process time, and eluent volume. The reference data can be obtained by recording any one of the molecular identity, molecular weight, molecular size, volume, chromatographic process time, and eluent volume of the reference molecule processed by the chromatographic apparatus 400.

[0065] In one example, the reference molecule is a useful protein with known molecular identity, known molecular weight, and known molecular size. The solution including the reference molecule is processed by the chromatographic apparatus 400, and the chromatographic process time and / or eluent volume are recorded and stored in a memory (e.g., in the memory of the control unit 410).

[0066] Step 620: Obtain the elution progress measurement and the corresponding quantitative measurement of the concentration of molecules in the eluent indicating the chromatographic equipment.

[0067] Elution progress can typically be measured, for example, the elapsed time of operation of the chromatography apparatus 400. Elapsed time is usually counted from the time the sample is initially injected into the column of the chromatography apparatus 400, for example... Figure 1 As shown in the figure. Alternatively or concurrently, the elution progress can typically be measured as the eluent volume. The eluent volume is typically counted from the time the sample is initially injected into the column of the chromatographic apparatus 400, for example, as... Figure 1 As shown in the image.

[0068] The corresponding quantitative measure is usually recorded at roughly the same time as the elution progress measure, and is therefore obtained at the time point corresponding to the elution progress measure.

[0069] Alternatively or concurrently, a quantitative measure is taken to indicate the ultraviolet absorption of the eluent.

[0070] Alternatively, the elution progress measurement indicates the running time of the elution process or the volume of the eluent.

[0071] Step 630: Estimate the size measure of the molecular size of the molecules in the eluent.

[0072] In one embodiment, molecular size is estimated by using a predetermined relationship included in reference data and by using a measure of elution progress.

[0073] In one embodiment, the predetermined relationship is indicated by a curve, and the predetermined relationship is obtained by performing curve fitting on a pair of values ​​including the gel phase distribution coefficient Kav and the corresponding molecular size, i.e., obtaining the curve.

[0074] In one example, the line equation (e.g., y = k*x + m) is fitted to value pairs, such as relative to... Figure 7 Further description.

[0075] In one embodiment, the elution progress measure is the elution volume Ve, where the gel phase distribution coefficient Kav depends on the total column volume Vt, the elution volume Ve, and the column void volume V0.

[0076] Alternatively or otherwise, the gel phase distribution coefficient Kav is given by the following relationship:

[0077] Kav = ((Ve-V0)) / ((Vt-V0)), where Ve is the elution volume, V0 is the column void volume, and Ve is the elution volume.

[0078] Alternatively or in an alternative location, the pre-determined relationship is a polynomial relationship.

[0079] Step 640: Render the representation of quantitative and magnitude measurements 300.

[0080] The representation of 300 may include a simplified diagram, such as relative to Figure 3 As stated above.

[0081] Alternatively, 300 may be represented as a simplified diagram having quantitative measures represented on a first axis and molecular sizes on a second axis, such as relative to... Figure 3 As stated above.

[0082] Step 650: Control the display to show the message to the user of the chromatography equipment.

[0083] In one example, the display 418 of the chromatography apparatus 400 is controlled to display representation 300 by sending a control signal from the processing circuitry system to the display 418.

[0084] In one embodiment, the method further includes using an estimated molecular size and one or more predetermined intervals of molecular size to activate or deactivate fractionation.

[0085] In one example, the outlet valve 420 is activated or deactivated by distributing the eluent to one or more outlets or outlet ports 421-423 in response to a control signal received from the control unit 410, for example, by using an estimated molecular size and one or more predetermined intervals of the molecular size.

[0086] Figure 7 Predetermined relationships according to one or more embodiments of this disclosure are shown. As previously stated relative to... Figure 6 The molecular size can be estimated by using a predetermined relationship included in the reference data and by using a measure of elution progress. The predetermined relationship can then be used to estimate a size measure indicating the molecular size of molecules in the eluent.

[0087] exist Figure 7 The graph shows a curve in the form of a line (700). The value pairs, including the gel phase distribution coefficient Kav and the corresponding molecular size, are shown as magnified black dots in the graph. The line curve is then fitted to these value pairs, for example using regression or any other suitable curve fitting technique.

[0088] It is envisioned that the curve described by the polynomial relation can be used alternatively for curve fitting of value pairs.

[0089] exist Figure 7 In the example, the relationship y = 0.3238x + 0.9683 has been obtained using value-pair-based curve fitting.

[0090] In one embodiment, a computer program is provided and includes computer-executable instructions for causing the control unit 410 to perform any method steps described herein when executed on a processing circuitry system included in the control unit 410.

[0091] In one embodiment, a computer program product is provided and includes a computer-readable storage medium having the aforementioned computer program embodied therein.

[0092] In embodiments, the communication network uses wired or wireless communication technologies for communication, including Local Area Network (LAN), Metropolitan Area Network (MAN), Global System for Mobile Communications (GSM), Enhanced Data GSM Environment (EDGE), Universal Mobile Telecommunications System (UMTS), Long Term Evolution (LTE), High Speed ​​Downlink Packet Access (HSDPA), Wideband Code Division Multiple Access (W-CDMA), Code Division Multiple Access (CDMA), Time Division Multiple Access (TDMA), Bluetooth®, Zigbee®, Wi-Fi, Voice over Internet Protocol (VoIP), LTE Advanced, IEEE 802.16m, WirelessMAN-Advanced, Evolved High Speed ​​Packet Access (HSPA+), 3GPP Long Term Evolution (LTE), Mobile WiMAX (IEEE 802.16e), Ultra Mobile Broadband (UMB) (formerly Evolved Data Optimized (EV-DO) Rev. C), Fast Low Latency Access with Seamless Handover Orthogonal Frequency Division Multiplexing (Flash-OFDM), High Capacity Space Division Multiple Access (iBurst®), and Mobile Broadband Wireless Access (MBWA) (IEEE 802.16m). It includes at least one of the following, but is not limited to: 802.20 system, high-performance radio metropolitan area network (HIPERMAN), wavelength division multiple access (BDMA), global microwave access interoperability (Wi-MAX), and ultrasonic communication.

[0093] Furthermore, as will be known to those skilled in the art, the control unit 410 may include the necessary communication capabilities, in the form of functions, components, units, elements, etc., for executing this solution. Other examples of such components, units, elements, and functions are: processors, memory, buffers, control logic, encoders, decoders, rate matchers, rate dematchers, mapping units, multipliers, decision units, selection units, switches, interleavers, deinterleavers, modulators, demodulators, inputs, outputs, antennas, amplifiers, receiver units, transmitter units, DSPs, MSDs, TCM encoders, TCM decoders, power supply units, power feeders, communication interfaces, communication protocols, etc., which are appropriately arranged together for executing this solution.

[0094] In particular, the processing circuitry system and / or processing components of this disclosure may include one or more instances of processing circuitry systems, processor modules, and multiple processors, central processing units (CPUs), processing units, processing circuits, processors, application-specific integrated circuits (ASICs), microprocessors, field-programmable gate arrays (FPGAs), or other processing logic capable of interpreting and executing instructions. Therefore, the expression "processing circuitry system" and / or "processing components" may refer to a processing circuitry system comprising multiple processing circuits, such as any, part, or all of the aforementioned processing circuitry systems. Processing components may further perform data processing functions for data input, output, and processing, including data buffering and device control functions, such as call processing control, user interface control, etc.

[0095] Finally, it should be understood that the present invention is not limited to the above embodiments, but also relates to and incorporates all embodiments within the scope of the appended independent claims.

Claims

1. A computer-implemented method (600) performed by a chromatography device (400) configured to separate molecules of varying sizes from an eluent, the method comprising: obtaining (610) reference data for a chromatography column of the chromatography apparatus, wherein the reference data indicates elution characteristics for a set of molecular sizes; obtaining (620) an elution progress measure and a corresponding quantitative measure indicative of the concentration of the molecule in the eluate of the chromatographic device; estimating (630) a size measure indicative of a molecular size of the molecule in the eluate; rendering (640) a representation (300) indicating the quantitative measure and the size measure; controlling (650) a display to display said representation to a user of said chromatographic device in substantially real time, Wherein, the molecular size is estimated using the predetermined relationship included in the reference data and the elution progress measure, and the predetermined relationship indicates a curve, and the predetermined relationship is obtained by performing curve fitting including a pair of values ​​of the gel phase distribution coefficient Kav and the corresponding molecular size.

2. The method (600) of claim 1, wherein: The elution progress measure is the elution volume Ve, wherein the gel phase distribution coefficient Kav depends on the total volume Vt of the column, the elution volume Ve and the column void volume V0.

3. The method (600) of claim 2, wherein: The gel phase distribution coefficient Kav is given by the following relationship:

4. The method (600) of claim 1 or 2, wherein: The predetermined relationship is a polynomial relationship.

5. The method (600) of claim 1 or 2, wherein: The quantitative measurement is indicative of the ultraviolet light absorbance of the eluate.

6. The method (600) of claim 1 or 2, wherein: The elution progress measure indicates the elution process run time or the eluate volume.

7. The method (600) of claim 1 or 2, wherein: The representation (300) is a simplified diagram having the quantitative measure represented on a first axis and the molecular size on a second axis.

8. The method (600) of claim 1 or 2, further comprising using the molecular size and one or more predetermined intervals of molecular size to activate or deactivate fractionation.

9. A chromatography device (400) configured to separate molecules of varying sizes from an eluent, the chromatography device (400) comprising: column(441); Display (411); and a control unit (410) comprising a circuit, the circuit comprising: Processing circuit (412); as well as A memory (415) comprising instructions executable by the processing circuit (412), whereby the chromatography device is configured to perform the method of any one of claims 1 to 8.

10. A computer-readable storage medium having a computer program stored thereon, the computer program comprising computer-executable instructions, the computer-executable instructions being used to cause the control unit (410) to perform the method steps (600) as described in any one of claims 1 to 8 when the computer-executable instructions are executed on a processing circuit included in the control unit (410).

11. A computer program product, comprising a computer program, wherein when the computer program is executed by a processor of a computer, the computer program implements the method steps according to any one of claims 1 to 8.

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