A chromatography column pH value adjustment system and method

Abstract

The invention discloses a chromatography column pH value regulating system and method, which relates to the technical field of chromatography and comprises an execution system, wherein the execution system comprises a raw material temporary storage tank, an acid tank, an alkali tank, a pH sensor, a main pump, a regulating pump, a flow meter M1, a flow meter M2, a valve T1, a valve T2, a valve T3 and a valve T4, wherein the raw material temporary storage tank is connected in series with the main pump, the main pump is connected in series with the pH sensor and the flow meter M1, the main pump is connected in series with the valve T1, the valve T1 is connected in series with the chromatography column, the bottom of the chromatography column is connected in series with a finished product collecting end, the top of the chromatography column is connected in series with a waste liquid pump, the acid tank is connected in series with the valve T2, the alkali tank is connected in series with the valve T4, the valve T2 and the valve T4 are both connected in series with the regulating pump, the regulating pump is connected in series with the valve T3, the valve T3 is connected in series with the flow meter M2 and the chromatography column, and the invention has the characteristic of dynamic adjustment.

Description

Technical Field

[0001] The invention relates to the technical field of chromatography, and in particular to a chromatography column pH value regulating system and method. Background Art

[0002] Chromatography may be used in the recovery of urine. Chromatography uses the different distribution and movement speeds of different substances in the stationary phase and mobile phase to separate the components in the mixture. The chromatography column consists of a tubular column filled with a stationary phase. The mobile phase enters the column from the top and flows out from the bottom after passing through the filler. Porous gel chromatography is a method of separating proteins such as urokinase in urine by gel column chromatography. The pH value affects the charge state between the biomolecules and the gel, and thus affects the interaction force. It is crucial to maintain the pH at a fixed value. The pH value is usually adjusted by mixing a dilute solution of hydrochloric acid and sodium oxychloride.

[0003] The existing adjustment method is to test the pH of urine in the urine storage tank, and then adjust the pH in each chromatography column to a suitable value. As the chromatography process proceeds, the protein gradually reacts with the surface charge of the porous gel in the chromatography column to cause it to be lost. At this time, the pH value needs to be gradually increased so that functional groups such as hydroxyl and amine groups can accept protons, form negative charges, and form electrostatic attraction with the porous gel surface to enhance the adsorption effect. This adjustment method does not take into account the fact that the appropriate pH value is dynamic, resulting in limited adsorption effect. Therefore, it is necessary to design a chromatography column pH adjustment system and method for dynamic adjustment. Summary of the invention

[0004] The object of the present invention is to provide a chromatography column pH adjustment system and method to solve the problems raised in the above background technology.

[0005] In order to solve the above technical problems, the present invention provides the following technical solutions: a chromatography column pH value adjustment system and method, including an execution system, the execution system including a raw material temporary storage tank, an acid tank, an alkali tank, a PH sensor, a main pump, a regulating pump, a flow meter M1, a flow meter M2, a valve T1, a valve T2, a valve T3, and a valve T4, the raw material temporary storage tank is connected to the main pump, the main pump is connected to the PH sensor and the flow meter M1, the main pump is connected to the valve T1, the valve T1 is connected to the chromatography column, the bottom of the chromatography column is connected to the finished product collection end, the top of the chromatography column is connected to the waste liquid pump, the acid tank is connected to the valve T2, the alkali tank is connected to the valve T4, the valve T2 and the valve T4 are both connected to the regulating pump, the regulating pump is connected to the valve T3, and the valve T3 is connected to the flow meter M2 and the chromatography column.

[0006] According to the above technical solution, it also includes a control system, which includes a protein weighing unit, a reaction ratio estimation module, a pH value correction module, an acid-base flow calculation module, a valve control module, a chromatography volume adjustment module, and a database, wherein the protein weighing unit is electrically connected to the reaction ratio estimation module, the reaction ratio estimation module is electrically connected to the pH value correction module and the chromatography volume adjustment module, the pH value correction module is electrically connected to the acid-base flow calculation module, the acid-base flow calculation module and the chromatography volume adjustment module are electrically connected to the valve control module, and the database is electrically connected to the protein weighing unit and the reaction ratio estimation module;

[0007] The protein weighing unit is installed at the finished product collection end and is used to measure the weight of the separated protein product. The reaction ratio estimation module is used to estimate the ratio of the reaction between the porous gel and the protein in the chromatography column according to the weight of the protein product. The pH value correction module is used to correct the suitable pH value of each chromatography column. The acid and alkali flow calculation module is used to calculate the pumping flow of the acid and alkali solution according to the corrected pH value. The valve control module is used to control the on and off of each valve. The chromatography volume adjustment module is used to calculate the raw material volume that needs to be pumped by each chromatography column during each chromatography. The database is used to store the weight of the protein chromatographed out of each chromatography column and the ratio of the reaction between the porous gel and the protein.

[0008] According to the above technical solution, the working method of the system is:

[0009] S1. When a specified amount of urine is stored in the raw material temporary storage tank, the pH value of the urine is tested using a pH sensor, and the pumping flow rates of the acid and alkali are calculated according to the preset pH value and the preset chromatography volume;

[0010] S2, selectively open one of valves T2 and T4, and open valve T3 in turn, use the regulating pump to pump the acid and alkali solution into each chromatography column, start the main pump and open each valve T1 in turn, and pump the raw material into each chromatography column;

[0011] S3, using flowmeter M1 and flowmeter M2 to measure the flow of the pumped raw materials, acid solution and alkali solution, and using the valve control module to close the valve T1 and valve T3 corresponding to each chromatography column when the amount is reached;

[0012] S4, waiting for the chromatography to end, starting the waste liquid pump to pump away the waste liquid, and using the protein weighing unit to weigh the weight of the chromatographic protein in each finished product collection end;

[0013] S5. estimating the ratio of the porous gel to the protein reaction according to the weight of the protein, and calculating the appropriate pH value of each chromatography column, and calculating the pumping flow rate of the acid and alkali solution required for each chromatography column according to the appropriate pH value;

[0014] S6. Repeat steps S2-S5, and calculate the volume of raw materials that need to be pumped for each chromatography column during each chromatography according to the weight of the protein, and then correct the flow rates of the acid and alkali solutions.

[0015] According to the above technical solution, in S1, the specific calculation method of the pumping flow rate of the acid solution and the alkali solution is as follows: the preset pH value is A0, the pH value of the urine is measured as A, when A<A0, it is necessary to open valve T4 to pump the alkali solution to increase the pH by A0-A, when A>A0, it is necessary to open valve T2 to pump the acid solution to reduce the pH by A-A0, the preset chromatography volume is V0, and the volumes of the acid solution and the alkali solution to be pumped are V1 and V2 respectively, the hydrogen ion concentration of the acid tank and the hydroxide ion concentration of the alkali tank are both amol / L, so the volume calculation formulas of the acid solution and the alkali solution to be pumped are respectively, the acid solution pH formula:

[0016]

[0017] Lye pH formula:

[0018]

[0019] The values ​​of V1 and V2, i.e. the pumping flow rates of the acid and alkali solutions, are obtained according to the formula.

[0020] According to the above technical solution, in S5, the specific method for estimating the ratio of the porous gel to the protein reaction is:

[0021] S5-1. When measuring after a chromatography, the protein weight measured in the finished product collection end is M i , where i is the serial number of the chromatography column. When the protein separation amount of a certain chromatography is lower than the critical value, that is, when the porous gel reaches the end of its service life, the proportion of porous gel that has not reacted with protein is k%. At this time, the total mass of separated protein is M0. Therefore, when measuring after a certain chromatography, the proportion of porous gel and protein reaction is

[0022] According to the above technical solution, in S5, the calculation method of the pumping flow rate of the acid solution and the alkali solution required for each chromatography column is:

[0023] S5-2. The appropriate pH increment for the next chromatography is Where μ is the conversion factor of the appropriate pH increment and the ratio of the porous gel to the protein reaction, and the appropriate pH value A i for A iInstead of A0, the acid pH formula and the alkali pH formula are substituted into S1 for calculation to obtain the pumping flow rates of the acid and alkali required for each chromatography column.

[0024] According to the above technical solution, in S6, the volume of raw materials that need to be pumped by each chromatography column during each chromatography is H i The calculation method is:

[0025] Calculate the total protein weight ∑M measured in the finished product collection end of all chromatography columns i , M i The larger the corresponding H i The total volume of raw materials distributed to all columns in each round of chromatography is ∑H i remains unchanged and is always the sum of the preset chromatography volumes, so δ is the raw material volume ratio coefficient.

[0026] According to the above technical solution, in S6, the specific method for correcting the flow rate of the acid solution and the alkali solution is: i By replacing V0 in the acid pH formula and the alkali pH formula for calculation, new values ​​of V1 and V2 corresponding to each chromatography column are obtained, that is, the corrected flow rates of the acid and alkali.

[0027] Compared with the prior art, the beneficial effects achieved by the present invention are as follows: the present invention measures the protein content separated by each chromatography column and estimates the ratio of the reaction between the porous gel and the protein in the chromatography column, thereby calculating the most suitable pH value for the current chromatography column, and dynamically adjusts the pH value of each chromatography column during each chromatography to enhance the adsorption effect, and separately regulates the volume of urine filtered by each chromatography column, thereby adjusting the pumping amount of the acid and alkali solution, so that the porous gel of each chromatography column reaches the expected service life at the same time as much as possible, avoids the reduction in utilization efficiency caused by partial disuse, and facilitates unified replacement. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] The accompanying drawings are used to provide a further understanding of the present invention and constitute a part of the specification. Together with the embodiments of the present invention, they are used to explain the present invention and do not constitute a limitation of the present invention. In the accompanying drawings:

[0029] Figure 1 It is a schematic diagram of the overall principle of the present invention;

[0030] Figure 2 It is a schematic diagram of the overall module structure of the present invention. DETAILED DESCRIPTION

[0031] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0032] See also Figure 1 and Figure 2 The present invention provides a technical solution: a chromatography column pH value regulating system and method, including an execution system, the execution system includes a raw material temporary storage tank, an acid tank, an alkali tank, a PH sensor, a main pump, a regulating pump, a flow meter M1, a flow meter M2, a valve T1, a valve T2, a valve T3, and a valve T4, the raw material temporary storage tank is connected to the main pump, the main pump is connected to the PH sensor and the flow meter M1, the main pump is connected to the valve T1, the valve T1 is connected to the chromatography column, the bottom of the chromatography column is connected to the finished product collection end, the top of the chromatography column is connected to the waste liquid pump, the acid tank is connected to the valve T2, the alkali tank is connected to the valve T4, the valves T2 and T4 are both connected to the regulating pump, the regulating pump is connected to the valve T3, and the valve T3 is connected to the flow meter M2 and the chromatography column;

[0033] It also includes a control system, which includes a protein weighing unit, a reaction ratio estimation module, a pH value correction module, an acid-base flow calculation module, a valve control module, a chromatography volume adjustment module, and a database. The protein weighing unit is electrically connected to the reaction ratio estimation module, the reaction ratio estimation module is electrically connected to the pH value correction module and the chromatography volume adjustment module, the pH value correction module is electrically connected to the acid-base flow calculation module, the acid-base flow calculation module and the chromatography volume adjustment module are electrically connected to the valve control module, and the database is electrically connected to the protein weighing unit and the reaction ratio estimation module;

[0034] The protein weighing unit is installed at the finished product collection end, and is used to measure the weight of the separated protein product. The reaction ratio estimation module is used to estimate the ratio of the reaction between the porous gel and the protein in the chromatography column according to the weight of the protein product. The pH value correction module is used to correct the suitable pH value of each chromatography column. The acid-base flow calculation module is used to calculate the pumping flow of the acid-base solution according to the corrected pH value. The valve control module is used to control the on-off of each valve. The chromatography volume adjustment module is used to calculate the raw material volume that needs to be pumped by each chromatography column during each chromatography. The database is used to store the weight of the protein chromatographically chromatographically obtained from each chromatography column and the ratio of the reaction between the porous gel and the protein.

[0035] The system works as follows:

[0036] S1. When a specified amount of urine is stored in the raw material temporary storage tank, the pH value of the urine is tested using a pH sensor, and the pumping flow rates of the acid and alkali are calculated according to the preset pH value and the preset chromatography volume;

[0037] S2, selectively open one of valves T2 and T4, and open valve T3 in turn, use the regulating pump to pump the acid and alkali solution into each chromatography column, start the main pump and open each valve T1 in turn, and pump the raw material into each chromatography column;

[0038] S3, using flowmeter M1 and flowmeter M2 to measure the flow of the pumped raw materials, acid solution and alkali solution, and using the valve control module to close the valve T1 and valve T3 corresponding to each chromatography column when the amount is reached;

[0039] S4, waiting for the chromatography to end, starting the waste liquid pump to pump away the waste liquid, and using the protein weighing unit to weigh the weight of the chromatographic protein in each finished product collection end;

[0040] S5. estimating the ratio of the porous gel to the protein reaction according to the weight of the protein, and calculating the appropriate pH value of each chromatography column, and calculating the pumping flow rate of the acid and alkali solution required for each chromatography column according to the appropriate pH value;

[0041] S6, repeating steps S2-S5, and calculating the volume of raw materials that need to be pumped for each chromatography column during each chromatography according to the weight of the protein, and then correcting the flow rates of the acid solution and the alkali solution;

[0042] In S1, the specific calculation method of the pumping flow of acid and alkali is as follows: the preset pH value is A0, and the pH value of urine is measured as A. When A<A0, valve T4 needs to be opened to pump alkali to increase the pH by A0-A. When A>A0, valve T2 needs to be opened to pump acid to reduce the pH by A-A0. The preset chromatography volume is V0. The volumes of acid and alkali to be pumped are V1 and V2 respectively. The hydrogen ion concentration of the acid tank and the hydroxide ion concentration of the alkali tank are both amol / L. Therefore, the volume calculation formulas of acid and alkali to be pumped are respectively: acid pH formula:

[0043]

[0044] Lye pH formula:

[0045]

[0046] According to the formula, the values ​​of V1 and V2, i.e. the pumping flow rates of the acid and alkali solutions, are obtained;

[0047] In S5, the specific method for estimating the ratio of porous gel to protein reaction is:

[0048] S5-1. When measuring after a chromatography, the protein weight measured in the finished product collection end is M i , where i is the serial number of the chromatography column. When the protein separation amount of a certain chromatography is lower than the critical value, that is, when the porous gel reaches the end of its service life, the proportion of porous gel that has not reacted with protein is k%. At this time, the total mass of separated protein is M0. Therefore, when measuring after a certain chromatography, the proportion of porous gel and protein reaction is

[0049] In S5, the calculation method for the pumping flow rate of acid and alkali required for each chromatography column is:

[0050] S5-2. The appropriate pH increment for the next chromatography is Where μ is the conversion factor of the appropriate pH increment and the ratio of the porous gel to the protein reaction, and the appropriate pH value A i for A i Substitute A0 into S1 to calculate the acid pH formula and alkali pH formula to obtain the pumping flow rate of acid and alkali required for each chromatography column;

[0051] In S6, the volume of raw material that needs to be pumped by each chromatography column during each chromatography is H i The calculation method is:

[0052] Calculate the total protein weight ∑M measured in the finished product collection end of all chromatography columns i , M i The larger the corresponding H i The total volume of raw materials distributed to all columns in each round of chromatography is ∑H i remains unchanged and is always the sum of the preset chromatography volumes, so δ is the raw material volume ratio coefficient;

[0053] In S6, the specific method for correcting the flow rate of acid and alkali is: i By replacing V0 in the acid pH formula and the alkali pH formula for calculation, new values ​​of V1 and V2 corresponding to each chromatography column are obtained, that is, the corrected flow rates of the acid and alkali.

[0054] Since urine of different depths will be pumped into the chromatography column each time the chromatography is performed, the protein content in the urine in each chromatography column will be slightly different. As multiple chromatography operations are accumulated, the separated protein content will be different. This will cause the porous gel in some chromatography columns to reach the end of its service life, while some have not been used up and cannot be replaced uniformly. Therefore, each time urine, i.e. raw material, is pumped, the chromatography column corresponding to the one with more separated protein will be given a smaller raw material volume for the next chromatography, which will reduce the difference in the degree of use of the porous gel.

[0055] By measuring the protein content separated by each chromatography column and estimating the ratio of the reaction between the porous gel and the protein in the chromatography column, the most suitable pH value for the current chromatography column can be calculated. The pH value of each chromatography column can be dynamically adjusted during each chromatography to enhance the adsorption effect. The volume of urine filtered by each chromatography column can be regulated separately, and then the pumping volume of the acid and alkali solution can be adjusted, so that the porous gel of each chromatography column can reach its expected service life at the same time as much as possible, avoiding the reduction in utilization efficiency caused by partial disuse and facilitating unified replacement.

[0056] It should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device.

[0057] Finally, it should be noted that the above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or replace some of the technical features therein by equivalents. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A chromatography column pH adjustment system, characterized in that: It comprises an execution system, which comprises a raw material temporary storage tank, an acid tank, an alkali tank, a PH sensor, a main pump, a regulating pump, a flow meter M1, a flow meter M2, a valve T1, a valve T2, a valve T3, and a valve T4. The raw material temporary storage tank is connected to the main pump, the main pump is connected to the PH sensor and the flow meter M1, the main pump is connected to the valve T1, the valve T1 is connected to a chromatography column, the bottom of the chromatography column is connected to a finished product collection end, the top of the chromatography column is connected to a waste liquid pump, the acid tank is connected to the valve T2, the alkali tank is connected to the valve T4, the valve T2 and the valve T4 are both connected to the regulating pump, the regulating pump is connected to the valve T3, and the valve T3 is connected to the flow meter M2 and the chromatography column; It also includes a control system, which includes a protein weighing unit, a reaction ratio estimation module, a pH value correction module, an acid-base flow calculation module, a valve control module, a chromatography volume adjustment module, and a database, wherein the protein weighing unit is electrically connected to the reaction ratio estimation module, the reaction ratio estimation module is electrically connected to the pH value correction module and the chromatography volume adjustment module, the pH value correction module is electrically connected to the acid-base flow calculation module, the acid-base flow calculation module and the chromatography volume adjustment module are electrically connected to the valve control module, and the database is electrically connected to the protein weighing unit and the reaction ratio estimation module; The protein weighing unit is installed at the finished product collection end, and is used to measure the weight of the separated protein product. The reaction ratio estimation module is used to estimate the ratio of the reaction between the porous gel and the protein in the chromatography column according to the weight of the protein product. The pH value correction module is used to correct the pH value suitable for each chromatography column. The acid-base flow calculation module is used to calculate the pumping flow of the acid-base solution according to the corrected pH value. The valve control module is used to control the on-off of each valve. The chromatography volume adjustment module is used to calculate the raw material volume that needs to be pumped by each chromatography column during each chromatography. The database is used to store the weight of the protein chromatographed out of each chromatography column and the ratio of the reaction between the porous gel and the protein. The system works as follows: S1. When a specified amount of urine is stored in the raw material temporary storage tank, the pH value of the urine is tested using a pH sensor, and the pumping flow rates of the acid and alkali are calculated according to the preset pH value and the preset chromatography volume; S2, selectively open one of valves T2 and T4, and open valve T3 in turn, use the regulating pump to pump the acid and alkali solution into each chromatography column, start the main pump and open each valve T1 in turn, and pump the raw material into each chromatography column; S3, using flowmeter M1 and flowmeter M2 to measure the flow of the pumped raw materials, acid solution and alkali solution, and using the valve control module to close the valve T1 and valve T3 corresponding to each chromatography column when the amount is reached; S4, waiting for the chromatography to end, starting the waste liquid pump to pump away the waste liquid, and using the protein weighing unit to weigh the weight of the chromatographic protein in each finished product collection end; S5. estimating the ratio of the porous gel to the protein reaction according to the weight of the protein, and calculating the appropriate pH value of each chromatography column, and calculating the pumping flow rate of the acid and alkali solution required for each chromatography column according to the appropriate pH value; S6. Repeat steps S2-S5, and calculate the volume of raw materials that need to be pumped for each chromatography column during each chromatography according to the weight of the protein, and then correct the flow rates of the acid and alkali solutions.

2. A chromatography column pH adjustment system according to claim 1, characterized in that: In S1, the specific calculation method of the pumping flow rate of the acid solution and the alkali solution is: the preset pH value is The pH value of urine was measured as ,when When the pH is high, valve T4 needs to be opened to pump alkali solution. ,when When the pH is lowered, valve T2 needs to be opened to pump acid. , the default chromatography volume is , assuming that the volumes of acid and alkali to be pumped are and The hydrogen ion concentration of the acid tank and the hydroxide ion concentration of the alkali tank are both Therefore, the volume calculation formulas for the acid and alkali solution that need to be pumped are: Acid PH formula: ； Lye pH formula: ； According to the formula and The value of is the pumping flow rate of acid and alkali solution.

3. A chromatography column pH adjustment system according to claim 2, characterized in that: In S5, the specific method for estimating the ratio of the porous gel to the protein reaction is: S5-1. When measuring after a chromatography, the protein weight measured in the finished product collection end is ,in is the serial number of the chromatography column. When the protein separation amount of a chromatography is lower than the critical value, that is, when the porous gel reaches the end of its service life, the proportion of the porous gel that has not reacted with the protein is , the total mass of separated protein is , so the ratio of porous gel to protein reaction after a chromatography is .

4. A chromatography column pH adjustment system according to claim 3, characterized in that: In S5, the calculation method of the pumping flow rate of the acid solution and the alkali solution required for each chromatography column is: S5-2. The appropriate pH increment for the next chromatography is ,in is the conversion factor of the appropriate pH increment and the ratio of the porous gel to the protein reaction, and the appropriate pH value for ,Will replace Substitute the acid pH formula and the alkali pH formula into S1 to calculate and obtain the pumping flow rates of the acid and alkali required for each chromatography column.

5. A chromatography column pH adjustment system according to claim 4, characterized in that: In S6, the volume of raw material that needs to be pumped by each chromatography column during each chromatography The calculation method is: Calculate the sum of the protein weights measured in the finished product collection end of all columns , The larger the corresponding The total volume of raw materials distributed to all columns in each round of chromatography decreases in direct proportion to remains unchanged and is always the sum of the preset chromatography volumes, so , , is the raw material volume ratio coefficient.

6. A chromatography column pH adjustment system according to claim 5, characterized in that: In S6, the specific method for correcting the flow rate of the acid solution and the alkali solution is: Replace the acid pH formula and the alkali pH formula Calculate and obtain the new corresponding to each column and The value is the corrected flow rate of acid and alkali solution.

Images