CN109276911A - A kind of continuous chromatography separation system and method - Google Patents
A kind of continuous chromatography separation system and method Download PDFInfo
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- CN109276911A CN109276911A CN201811443755.3A CN201811443755A CN109276911A CN 109276911 A CN109276911 A CN 109276911A CN 201811443755 A CN201811443755 A CN 201811443755A CN 109276911 A CN109276911 A CN 109276911A
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- 238000004587 chromatography analysis Methods 0.000 title claims abstract description 40
- 238000000926 separation method Methods 0.000 title claims abstract description 26
- 238000000034 method Methods 0.000 title description 8
- 238000001802 infusion Methods 0.000 claims abstract description 35
- 239000012535 impurity Substances 0.000 claims description 15
- 230000008676 import Effects 0.000 claims description 12
- 238000010828 elution Methods 0.000 claims description 4
- 238000004458 analytical method Methods 0.000 claims description 3
- 238000007689 inspection Methods 0.000 claims 1
- 238000002955 isolation Methods 0.000 abstract description 4
- 102000004169 proteins and genes Human genes 0.000 abstract description 4
- 108090000623 proteins and genes Proteins 0.000 abstract description 4
- 239000012071 phase Substances 0.000 description 52
- 239000000523 sample Substances 0.000 description 17
- 238000010586 diagram Methods 0.000 description 7
- 239000007791 liquid phase Substances 0.000 description 4
- 238000001042 affinity chromatography Methods 0.000 description 2
- 239000002808 molecular sieve Substances 0.000 description 2
- URGAHOPLAPQHLN-UHFFFAOYSA-N sodium aluminosilicate Chemical compound [Na+].[Al+3].[O-][Si]([O-])=O.[O-][Si]([O-])=O URGAHOPLAPQHLN-UHFFFAOYSA-N 0.000 description 2
- 238000007445 Chromatographic isolation Methods 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 229960000074 biopharmaceutical Drugs 0.000 description 1
- 238000013375 chromatographic separation Methods 0.000 description 1
- 238000011210 chromatographic step Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
- B01D15/14—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to the introduction of the feed to the apparatus
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
- B01D15/18—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to flow patterns
- B01D15/1864—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to flow patterns using two or more columns
Abstract
The invention discloses a kind of continuous chromatography separation systems, infusion device including being sequentially connected in series setting, multiple flow path selection valves and discharge port, further include and the one-to-one chromatographic column of flow path selection valve, the both ends of each chromatographic column are connected to the same flow path selection valve, each flow path selection valve can be toggled in state one and state two, state one: flow path is without chromatographic column, the outlet end of flow path selection valve is directly connected to its arrival end, state two: flow path passes through chromatographic column, the outlet end of flow path selection valve is connect with its arrival end by the upper and lower ends of chromatographic column.The application switches flow path by using flow path selection valve, therefore can continuously carry out a variety of isolation technics, shortens disengaging time, improves the separative efficiency of target protein.
Description
Technical field
The present invention relates to chromatography field, in particular to a kind of continuous chromatography separation system and method.
Background technique
In bio-pharmaceuticals process flow, needed in the downstream separation technique of biological sample using a variety of chromatographic separation technologies
Sequential working, for example first time separation first is carried out with affinity chromatography to a kind of target protein object, being collected into after product again
Into tomographic system, then second being carried out with the chromatography of ions and is separated, the product for collecting second of separation is again introduced into chromatography system
System can just be collected into target product after carrying out third time separation using molecular sieve, off-line type chromatographic isolation, not only production cost
Height, and low efficiency.
In the prior art, it requires to collect target protein object again after chromatography every time, the formula that is initially separated needs weight again
New addition product, therefore often separation takes a long time, therefore how to realize continuous chromatography to shorten disengaging time
The problem of being this field urgent need to resolve.
Summary of the invention
One kind of layer multiple chromatography time can be effectively shortened the technical problem to be solved in the present invention is to provide one kind
Continuous chromatography separation system.
In order to solve the above technical problem, the present invention provides technical solution be: a kind of continuous chromatography separation system, including
It is sequentially connected in series the infusion device, multiple flow path selection valves and discharge port of setting, further includes a pair of with the flow path selection valve one
The chromatographic column answered, each flow path selection valve control the connected state of corresponding chromatographic column, each flow path selection valve
It is toggled in state one and state two, infusion device, flow path selection valve, at least one layer of analysis column and discharge port form a flow path；
The state one: the outlet end of flow path selection valve is directly connected to its arrival end, and flow path selection valve controls chromatographic column
It is disconnected with the flow path,
The state two: the outlet end of flow path selection valve is connect with its arrival end by the both ends of chromatographic column, flow path selection
Valve control chromatographic column is connected in the flow path.
It is specific:
The state one: the inlet end and an outlet end connection of flow path selection valve, the upper and lower ends of corresponding chromatographic column
Connection,
The state two: the input end of flow path selection valve is connected to the upper end of chromatographic column, the lower end of the chromatographic column and institute
State the outlet end connection of flow path selection valve.
Further, being additionally provided with detector on the discharge port.
Further, the infusion device is made of at least one infusion pump, the infusion pump is plunger pump or diaphragm
Pump.
Further, the infusion device is also connect with multiple mobile phase entrances, the number of the mobile phase entrance is extremely
One more than few quantity than the chromatographic column.
Further, the mobile phase entrance includes four, the flow path selection valve includes three, the chromatographic column packet
Include three.
Further, including the first mobile phase entrance being connected to the infusion device, the second mobile phase entrance and
Three mobile phase entrances, the 4th mobile phase entrance and sample inlet, the input end of the infusion device and first flow path selector valve
Connection, the first flow path selector valve are also connected to the upper and lower ends of first chromatographic column, the first flow path selector valve
Outlet end is connected to the input end of second flow path selector valve, upper and lower ends of the second flow path selector valve also with the second chromatographic column
Connection, the outlet end of the second flow path selector valve are connected to the input end of third flow path selection valve, the third flow path selection
Valve is also connected to the upper and lower ends of third chromatographic column, the outlet end discharge port connection of the third flow path selection valve.
Further, the flow path selection valve includes input end, outlet end, chromatographic column import and chromatographs column outlet,
The chromatographic column import and the chromatography column outlet are respectively used to connect with the upper and lower ends of same chromatographic column, the flow path selection
It is provided with rotor in valve, flow path selection valve can be made to switch in state one and state two by rotor.
The application further provides a kind of continuous chromatography method, and used technical solution is: the following steps are included:
S1, state two is set by connect with infusion device first flow path selection valve, other flow path selection valves are set
It is set to state one, injects a sample into system, first chromatographic column corresponding with first flow path selection valve captures target components, and
The impurity in target components is eluted afterwards；
S2, state two is set by (n+1)th flow path selection valve, target components is eluted from n-th of series of strata column,
And target components are captured by (n+1)th chromatographic column, then, n-th of flow path selection valve is switched into state one, elutes target components
In impurity, wherein n indicate previous step in flow path selection valve serial number；
S3, step S2 is repeated until after the last one chromatographic column capture target components, by target components from the last one layer
It is eluted on analysis column, target components is detected and are collected.
Further, S1, set first flow path selector valve to state two, second flow path selector valve and the choosing of third flow path
It selects valve and is set as state one, inject a sample into system, the first chromatographic column captures target components, then elutes target components
In impurity；
S2, state two is set by second flow path selector valve, target components is eluted from the first chromatographic column, second
Chromatographic column captures target components, then, sets state one for first flow path selector valve, elutes miscellaneous in target components
Matter；
S3, state two is set by third flow path selection valve, target components is eluted from the second chromatographic column, third
Chromatographic column captures target components, then sets state two for second flow path selector valve, elutes miscellaneous in target components
Matter,
S4, the target components on third chromatographic column are eluted, the target components after elution is detected and collected.
The beneficial effects of the present invention are: the application switches flow path by using flow path selection valve, to realize on-line continuous
A variety of isolation technics, shorten disengaging time, improve the separative efficiency of target protein, and flow path selection valve passes through motor and drives
It is dynamic, so as to realize automation switching.
Detailed description of the invention
Fig. 1 is that flow path selection valve is in state schematic diagram for the moment.
Fig. 2 is schematic diagram when flow path selection valve is in state two.
Fig. 3 is the schematic diagram that first flow path selector valve is in state two.
Fig. 4 is the schematic diagram that first flow path selector valve and second flow path selector valve are in state two.
Fig. 5 is the schematic diagram that second flow path selector valve is in state two.
Fig. 6 is the schematic diagram that second flow path selector valve and third flow path selection valve are in state two.
Fig. 7 is the schematic diagram that third flow path selection valve is in state two.
Label is in figure: the first mobile phase entrance 11, the second mobile phase entrance 12, third mobile phase entrance the 13, the 4th flow
Dynamic phase entrance 14, sample inlet 2, infusion device 3, flow path selection valve 4, input end a, outlet end b, chromatographic column import d, chromatographic column
Export c, first flow path selector valve 41, second flow path selector valve 42, third flow path selection valve 43, chromatographic column 5, the first chromatographic column
51, the second chromatographic column 52, third chromatographic column 53, detector 6.
Specific embodiment
The present invention will be further explained below with reference to the attached drawings and specific examples, so that those skilled in the art can be with
It more fully understands the present invention and can be practiced, but illustrated embodiment is not as a limitation of the invention.
As shown in Figures 1 to 7, a kind of continuous chromatography separation system, including multiple mobile phase entrances, a sample inlet 2,
Mobile phase entrance and sample are connected to infusion device 3, and infusion device 3, multiple flow path selection valves 4 and discharge port pass sequentially through
Placed in series can also wherein install the detector 6 for detecting target components, in this application, the flow path on discharge port
Selector valve 4 is identical as 5 number of chromatographic column, and the number of the number wall chromatographic column 5 of mobile phase entrance at least has more one, chromatographic column 5
It is corresponded with flow path selection valve 4, i.e., the both ends of each chromatographic column 5 are only connected to a flow path selection valve 4, each stream
Road selector valve 4 can be toggled in state one and state two.
As shown in Figure 1, the state one: flow path is without chromatographic column 5, the outlet end a and its arrival end of flow path selection valve 4
B is directly connected to, specifically, the input end a of flow path selection valve 4 is connected to outlet end b, up and down the two of corresponding chromatographic column 5
End connection,
As shown in Fig. 2, the state two: flow path passes through chromatographic column 5, the outlet end a and its arrival end b of flow path selection valve 4
It is connected by the upper and lower ends of chromatographic column, specifically, the input end a of flow path selection valve 4 is connected to the upper end of chromatographic column 5, it is described
The lower end of chromatographic column 5 is connected to the outlet end b of the flow path selection valve 4.
Wherein, the access that flow path, that is, mobile phase is formed via the discharge port of infusion device flowing.
Specifically, the flow path selection valve 4 include at least four positions, be respectively input end a, outlet end b, chromatographic column into
Mouth d and chromatography column outlet c, the chromatographic column import d and the chromatography column outlet c are respectively used to upper with same chromatographic column 5
The connection of lower both ends, input end a are used for feed liquor, and outlet end b is used for drain, are provided with rotor in the flow path selection valve 4, rotor with
Motor connection, is rotated by motor driven rotor, so as to so that flow path selection valve 4 is switched in state one and state two,
It should be understood that flow path selection valve 4 can also use the selector valve of five positions or seven positions in other embodiments,
Chromatographic column 5 is linked into system flow as long as can enough be realized by switching flow path, and chromatographic column 5 is isolated in system stream
Outside road.
When being switched to state a period of time, input end a is connected to outlet end b, and the import of series of strata column is connected to series of strata column outlet, this shape
Under state, chromatographic column 5 is in the bypass of system, injects liquid phase at this time, and liquid phase may not flow into chromatographic column 5.
When the state two of being switched to, input end a is connected to chromatographic column import d, and chromatography column outlet c is connected to outlet end b, this
When inject liquid phase, liquid phase can be entered in chromatographic column 5 by input end a.
In this application, an infusion pump composition is included at least in the infusion device 3, preferably includes one or two
Infusion pump forms infusion device 3, wherein the infusion pump is plunger pump or diaphragm pump, and plunger pump optimizes for laboratory process
Or technique amplification stage, diaphragm pump are used for the large-scale production stage.
A kind of continuous chromatography separation method based on continuous chromatography separation system, comprising the following steps:
S1, state two, other flow path selection valves 4 are set by first flow path selection valve 4 connecting with infusion device 3
It is set as state one, the first chromatographic column 51 is in the flow path of sample, and other chromatographic columns 5 are in bypass, and sample is passed through
Sample inlet 2 injects infusion device 3, and infusion device 3 injects a sample into system, at this point, sample passes through first flow path selector valve 41
It enters in the first chromatographic column 51, first chromatographic column 5 captures the target components in sample, then passes through mobile phase entrance
The mobile phase for being used to elute impurity in target components is injected into system, to elute the impurity in target components.
S2, state two is set by (n+1)th flow path selection valve 4, n-th and (n+1)th chromatographic column 5 are in and are at this time
It unites in flow path, the mobile phase for being used to elute target components from n-th of chromatographic column 5 is injected by system by mobile phase entrance
Target components are eluted from n-th of series of strata column, are eluted the target components exerted oneself and enter (n+1)th by system, the mobile phase
In a chromatographic column 5, and target components is captured by (n+1)th chromatographic column 5, n-th of flow path selection valve 4 is then switched into shape
State one, only (n+1)th chromatographic column 5 is in system flow path at this time, will be used to elute target components by mobile phase entrance
The mobile phase of middle impurity is injected into system, elutes the impurity in target components, wherein n indicates flow path choosing in previous step
Select the serial number of valve 4；
S3, step S2 is repeated until the last one chromatographic column 5 completes above-mentioned steps, only have the selection of the last one flow path at this time
Valve 4 is in state two, therefore only the last one chromatographic column 5 is in system flow path, is then injected by mobile phase entrance
Mobile phase for eluting target components from the last one chromatographic column 5, the mobile phase by target components from the last one
It being eluted on chromatographic column 5, target components flow to discharge port, and device 6 after testing, and detector 6 detects target components,
When target components be required group of timesharing, target components are collected.
In order to make it easy to understand, the application provides a kind of preferred embodiment:
In the present embodiment, the mobile phase entrance includes four, the flowing of respectively the first mobile phase entrance 11, second
Phase entrance 12, third mobile phase entrance 13 and the 4th mobile phase entrance 14；The flow path selection valve 4 includes three, respectively the
One flow path selection valve 41, second flow path selector valve 42 and third flow path selection valve 43；The chromatographic column 5 includes three, respectively
For the first chromatographic column 51, the second chromatographic column 52 and third chromatographic column 53, wherein chromatographic column 5 can be respectively adopted affinity chromatography,
Ion chromatography and molecular sieve.
First mobile phase entrance 11, the second mobile phase entrance 12, third mobile phase entrance 13 and the 4th mobile phase entrance 14
And sample inlet 2 is connected to infusion device 3, the infusion device 3 is connected to the input end a of first flow path selector valve 41,
Chromatographic column import d and chromatography column outlet c in the first flow path selector valve 41 is respectively and above and below first chromatographic column 51
Both ends connection, the outlet end b of the first flow path selector valve 41 is connected to the input end a of second flow path selector valve 42, and described the
Chromatographic column import d in two flow path selection valves 42 and chromatography column outlet c are connected to the upper and lower ends of the second chromatographic column 52, and described the
The outlet end b of two flow path selection valves 42 is connected to the input end a of third flow path selection valve 43, in the third flow path selection valve 43
Chromatographic column import d and chromatography column outlet c be connected to the upper and lower ends of third chromatographic column 53, the third flow path selection valve 43
Outlet end b connect with detector 6.
In addition, also specifically providing a kind of chromatographic step based on above-mentioned continuous chromatography system:
S1, first flow path selector valve 41 is set to state two, second flow path selector valve 42 and third flow path selection valve 43
It is set as state one, at this point, the first chromatographic column 51 is in system flow path, the second chromatographic column 52 and third chromatographic column 53 are in
Sample is injected infusion device 3 by sample inlet 2 by bypass, and infusion device 3 injects a sample into first flow path selector valve 41, sample
Product enter in the first chromatographic column 51, and the first chromatographic column 51 captures target components, are infused by the first mobile phase entrance
Enter No.1 mobile phase, No.1 mobile phase is used to elute the impurity in target components；
S2, state two is set by second flow path selector valve 42, at this time first flow path selector valve 41 and second flow path selection
Valve 42 is in state two, injects No. two mobile phases by the second mobile phase entrance 12 at this time, and No. two mobile phases are used for target
Component is eluted from the first chromatographic column 51, and target components are all brought into the second chromatographic column 52 by No. two mobile phases,
Second chromatographic column 52 captures target components, then, sets state one for first flow path selector valve 41, at this point, only
Second chromatographic column 52 is in system flow path, is injected No.1 mobile phase by the first mobile phase entrance 11, is eluted target components
In impurity.
S3, state two is set by third flow path selection valve 43, at this time second flow path selector valve 42 and the selection of third flow path
Valve 43 is in state two, injects No. three mobile phases by third mobile phase entrance 13, No. three mobile phases are used for target components
It is eluted from the second chromatographic column 52, No. three mobile phases drive target components to enter third chromatographic column 53, third chromatographic column
53 pairs of target components capture, and then set state two for second flow path selector valve 42, at this point, only third chromatographic column 53
In system flow path, No.1 mobile phase is injected by the first mobile phase entrance 11, elutes the impurity in target components.
S4, No. four mobile phases are being injected by No. four mobile phase entrances, No. four mobile phases are used for third chromatographic column 53
Target components eluted, the target components after elution enter discharge gate, and by detector 6, detector 6 is to target components
It is detected, after detection target components are met the requirements, the target components after elution is collected.
Switch flow path by using flow path selection valve 4, therefore be capable of the kind isolation technics of on-line continuous, thus to target
Albumen carries out chromatography, the application can be made to be fully automated during the separation process by motor driven, pass through the application
A variety of isolation technics are carried out, separative efficiency efficiency improves three to four-fold.
Embodiment described above is only to absolutely prove the preferred embodiment invented and lifted, and the protection scope of invention is not
It is limited to this.Those skilled in the art made equivalent substitute or transformation on the basis of invention, in the protection model of invention
Within enclosing.The protection scope of invention is subject to claims.
Claims (9)
1. a kind of continuous chromatography separation system, which is characterized in that infusion device, the selection of multiple flow paths including being sequentially connected in series setting
Valve and discharge port further include controlling phase with the one-to-one chromatographic column of the flow path selection valve, each flow path selection valve
The connected state of chromatographic column is answered, each flow path selection valve can be toggled in state one and state two, infusion device, stream
Road selector valve, at least one layer of analysis column and discharge port form a flow path；
The state one: the outlet end of flow path selection valve is directly connected to its arrival end, and flow path selection valve controls chromatographic column and institute
Flow path disconnection is stated,
The state two: the outlet end of flow path selection valve is connect with its arrival end by the both ends of chromatographic column, flow path selection valve control
Chromatographic column processed is connected in the flow path.
2. a kind of continuous chromatography separation system as described in claim 1, which is characterized in that be additionally provided with inspection on the discharge port
Survey device.
3. a kind of continuous chromatography separation system as described in claim 1, which is characterized in that the infusion device is by least one
Infusion pump composition, the infusion pump are plunger pump or diaphragm pump.
4. a kind of continuous chromatography separation system as claimed in claim 3, which is characterized in that the infusion device also with multiple streams
Dynamic phase entrance connection, the number of the mobile phase entrance is at least one more than the quantity of the chromatographic column.
5. a kind of continuous chromatography separation system as claimed in claim 4, which is characterized in that the mobile phase entrance includes four
A, the flow path selection valve includes three, and the chromatographic column includes three.
6. a kind of continuous chromatography separation system as claimed in claim 5, which is characterized in that including being connected to the infusion device
The first mobile phase entrance, the second mobile phase entrance, third mobile phase entrance, the 4th mobile phase entrance and sample inlet, institute
Infusion device is stated to be connected to the input end of first flow path selector valve, the first flow path selector valve also with first chromatographic column
Upper and lower ends connection, the outlet end of the first flow path selector valve are connected to the input end of second flow path selector valve, and described second
Flow path selection valve is also connected to the upper and lower ends of the second chromatographic column, and the outlet end of the second flow path selector valve and third flow path select
The input end connection of valve is selected, the third flow path selection valve is also connected to the upper and lower ends of third chromatographic column, the third flow path
The outlet end of selector valve is connected to discharge port.
7. a kind of continuous chromatography separation system as described in claim 1, which is characterized in that the flow path selection valve includes import
End, outlet end, chromatographic column import and chromatography column outlet, the chromatographic column import and the chromatography column outlet are respectively used to and together
The upper and lower ends of one chromatographic column connect, and are provided with rotor in the flow path selection valve, flow path can be made to select by rotor
Valve is switched in state one and state two.
8. a kind of continuous chromatography separation method based on any one of claim 1 to 7 continuous chromatography separation system, including following step
It is rapid:
S1, state two is set by connect with infusion device first flow path selection valve, other flow path selection valves are set as
State one, injects a sample into system, and first chromatographic column corresponding with first flow path selection valve captures target components, then washes
Impurity in de- target components；
S2, state two is set by (n+1)th flow path selection valve, target components is eluted from n-th of series of strata column, and by
(n+1)th chromatographic column captures target components, then, n-th of flow path selection valve is switched to state one, is eluted in target components
Impurity, wherein n indicates the serial number of flow path selection valve in previous step；
S3, after repeating step S2 until the last one chromatographic column completes above-mentioned steps, by target components from the last one chromatographic column
On elute, target components are collected.
9. a kind of continuous chromatography separation method of the continuous chromatography separation system based on claim 6, comprising the following steps:
S1, first flow path selector valve is set to state two, second flow path selector valve and third flow path selection valve are set as state
One, system is injected a sample into, the first chromatographic column captures target components, then elutes the impurity in target components；
S2, state two is set by second flow path selector valve, target components is eluted from the first chromatographic column, the second chromatography
Column captures target components, then, sets state one for first flow path selector valve, elutes the impurity in target components；
S3, state two is set by third flow path selection valve, target components is eluted from the second chromatographic column, third chromatography
Column captures target components, then sets state two for second flow path selector valve, elutes the impurity in target components,
S4, the target components on third chromatographic column are eluted, the target components after elution is collected.
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| CN105664527A (en) * | 2016-01-27 | 2016-06-15 | 李广良 | Continuous chromatographic separation device and method using device |
| CN209361909U (en) * | 2018-11-29 | 2019-09-10 | 亿达康瑞生物科技（苏州）有限公司 | A kind of continuous chromatography separation system |
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