CN102258885B - Automatic separation system for integrated column chromatography - Google Patents
Automatic separation system for integrated column chromatography Download PDFInfo
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- CN102258885B CN102258885B CN 201110129336 CN201110129336A CN102258885B CN 102258885 B CN102258885 B CN 102258885B CN 201110129336 CN201110129336 CN 201110129336 CN 201110129336 A CN201110129336 A CN 201110129336A CN 102258885 B CN102258885 B CN 102258885B
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- 238000000926 separation method Methods 0.000 title claims abstract description 18
- 238000004440 column chromatography Methods 0.000 title abstract 2
- 239000007788 liquid Substances 0.000 claims abstract description 64
- 230000002572 peristaltic effect Effects 0.000 claims abstract description 28
- 238000005086 pumping Methods 0.000 claims abstract description 15
- 238000006243 chemical reaction Methods 0.000 claims abstract description 4
- 239000012530 fluid Substances 0.000 claims description 54
- 239000000523 sample Substances 0.000 claims description 54
- 238000001514 detection method Methods 0.000 claims description 25
- 238000004587 chromatography analysis Methods 0.000 claims description 22
- 239000003480 eluent Substances 0.000 claims description 21
- 238000004891 communication Methods 0.000 claims description 12
- 238000012360 testing method Methods 0.000 claims description 11
- 239000002699 waste material Substances 0.000 claims description 10
- 238000002156 mixing Methods 0.000 claims description 4
- 238000002347 injection Methods 0.000 claims description 3
- 239000007924 injection Substances 0.000 claims description 3
- 239000004973 liquid crystal related substance Substances 0.000 claims description 3
- 239000002994 raw material Substances 0.000 claims description 3
- 238000013519 translation Methods 0.000 claims description 3
- 238000005070 sampling Methods 0.000 abstract 3
- 239000011365 complex material Substances 0.000 abstract 2
- 244000005700 microbiome Species 0.000 abstract 2
- 239000000463 material Substances 0.000 description 20
- 230000000813 microbial effect Effects 0.000 description 12
- 239000012634 fragment Substances 0.000 description 9
- 238000000034 method Methods 0.000 description 9
- 239000007853 buffer solution Substances 0.000 description 3
- 238000011010 flushing procedure Methods 0.000 description 3
- 239000012535 impurity Substances 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 230000008929 regeneration Effects 0.000 description 3
- 238000011069 regeneration method Methods 0.000 description 3
- 239000007790 solid phase Substances 0.000 description 3
- 239000007791 liquid phase Substances 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 238000011017 operating method Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000005377 adsorption chromatography Methods 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
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Abstract
The invention discloses an automatic separation system for integrated column chromatography, comprising a mechanical part and a control part, wherein the mechanical part at least comprises a sampling and peristaltic pumping module, an integrated bedpost, a detector and a desampling and product-collecting module; the control part comprises a singlechip, an input/output unit, a relay and a driver; a sampling opening of the integrated bedpost is communicated with a peristaltic pump A in the peristaltic pumping module, a desampling opening and a liquid-collecting tank in the desampling and product-collecting module; the detector is arranged between a desampling opening of the integrated bedpost and the desampling and product-collecting module; and the singlechip is used for controlling the sampling and peristaltic pumping module and the desampling and product-collecting module and collecting and conversing a signal sent from the detector through an A/D (Analog/Digital) conversion chip. The system provided by the invention allows complex material liquid containing microorganism cells to pass through directly and can automatically treat the complex material liquid containing the microorganism cells or cell debris at high speed.
Description
Technical field
The present invention relates to the automatic chromatography of a kind of integral post system, belong to biology, chemical industry and field of mechanical technique.
Background technology
The integral post chromatography of macroporous type (also claiming integral bed or continuous bed chromatography) is the new bio separation method that grew up in recent years, the aperture of its integral post separating medium can reach several microns even hundreds of micron, allow to contain the microbial cell of submicron-scale or the complicated material liquid of cell fragment directly passes bed, the adsorption chromatography of object in this class separating medium separates rapidly, can be used for directly from containing the complicated material liquid separate targets biomolecular material of solid phase impurity such as microbial cell or cell fragment.This method has the overall merit of expansion bed and conventional fixed bed chromatography, can with filter, centrifugal, concentrated and chromatography several steps is integrated in step realization, the treating capacity of unit interval is big, the preparation type that is suitable for biochemical substances separates fast.Therefore, aspect the separation and purification of microbial fermentation biochemicals, natural product extraction or genetically engineered drug etc., have broad application prospects.
Though have many automatic separation systems that are used for biological chromatography both at home and abroad, fixed bed analysis system and highly effective liquid phase chromatographic system as expansion bed tomographic system, conventional mesolow, but, except the expansion bed system, the existing tomographic system overwhelming majority separates at analyzing the less preparation of separation or scale, and the complicated feed liquid fluid that does not allow to contain solid matters such as microbial cell or cell fragment is difficult to directly complex fluid be carried out quick separating treatment by column.The present both at home and abroad Special Automatic tomographic system of still not separating at the macroporous type integral post, the tomographic system of existing highly effective liquid phase chromatographic system or assembling, operation inconvenience, inapplicable or have a certain difficulty when mass preparation is separated.
Summary of the invention
For solving the aforementioned problems in the prior, the invention provides a kind of integral post chromatography automatic separation system, the complicated feed liquid that system of the present invention allows to contain microbial cell is directly passed through, and can direct High-Speed Automatic processing contains the complicated feed liquid of microbial cell or cell fragment.
For reaching above-mentioned purpose, the present invention adopts following technical scheme: a kind of integral post chromatography automatic separation system, formed by mechanical part and control section, mechanical part comprise at least sample introduction with wriggling pumping module, whole column, detector, go out sample and product collection module, control section is core with the single-chip microcomputer, comprise input-output unit, two above relays and three above drivers;
Sample introduction comprises four above sample control valves with wriggling pumping module, three above peristaltic pumps, six above fluid reservoirs and blender, the fluid reservoir that wherein is used for each eluent of storage is communicated with blender by the corresponding peristaltic pump that is used for this eluent of pumping respectively, blender is communicated with the fluid reservoir A that is used for storage mixing eluent, by being used for carrying the peristaltic pump A of liquid to comprise fluid reservoir A at least with the fluid reservoir that whole column injection port is communicated with to whole column, the fluid reservoir B that is used for storage of raw materials liquid, the fluid reservoir D that is used for storing the fluid reservoir C of equilibrium liquid and is used for store regenerated liquid is at fluid reservoir A, fluid reservoir B, be provided with sample control valve separately between fluid reservoir C and fluid reservoir D and the peristaltic pump A;
Go out that sample comprises with the product collection module going out the sample control valve more than three, be used for collecting object and the liquid collecting jar of waste liquid more than three, whole column outlet is communicated with the liquid collecting jar that respectively is used for collecting object and waste liquid, at whole column outlet and respectively be provided with out the sample control valve separately between the liquid collecting jar for collection object and waste liquid;
Detector is located at whole column outlet and is respectively gone out between the sample control valve;
Wherein, single-chip microcomputer is used for driving the driver of each peristaltic pump of control and controls sample introduction and wriggling pumping module for the relay A that drives sample control valve by driving, single-chip microcomputer is controlled out sample and product collection module by the relay B that drive to be used for driving the sample control valve, and output signal from detector is gathered and changed to single-chip microcomputer by the A/D conversion chip.
Single-chip microcomputer drives be used to the driver that drives each peristaltic pump of control respectively, is used for driving the relay A of each sample control valve with its drive circuit that carries, and is used for the relay B that driving respectively goes out the sample control valve.
In described control section, input-output unit comprises host computer, touch-screen, LCDs, host computer is by serial ports and single chip communication, touch-screen is connected with the I/O interface of single-chip microcomputer by touch-screen digital translation chip, and LCDs is connected with the I/O interface of single-chip microcomputer by the liquid crystal display driver.
Used detector is Ultraviolet Detector.
Also be provided with sensing module in the mechanical part, described sensing module comprises temperature sensor at least, two above liquid-pressure pick-ups, two above electrical conductivity unit and two above pH test cells, wherein be communicated with blender also and single chip communication by pipeline for detection of the electrical conductivity unit that mixes the eluent electrical conductivity and for detection of the pH test cell that mixes eluent pH value, electrical conductivity unit for detection of liquid electric conductivity in the integral post, for detection of the liquid-pressure pick-up of the fluid pressure of carrying to integral post and for detection of the temperature sensor of fluid temperature in the whole column is communicated with the outlet of peristaltic pump A by pipeline and and single chip communication, the pH test cell that reaches for detection of whole column liquid pH value for detection of the liquid-pressure pick-up of whole column fluid pressure is communicated with the outlet of whole column also and single chip communication by pipeline.
Described sensing module also comprises the air pressure probe for detection of the air capacity in the system.
Compared with prior art, integral post chromatography automatic separation system provided by the invention has the following advantages:
1. the separating medium aperture of native system can reach several microns even hundreds of micron, can allow to contain the microbial cell of submicron-scale or the complicated material liquid of cell fragment directly passes bed, can be used for directly separate targets biomolecular material from the complicated material liquid that contains solid phase impurity such as microbial cell or cell fragment.
2. the unit interval feed liquid treating capacity of native system is big, separates rapidly, and directly quickly separating target biomolecular material from complicated nutrient solution, zymotic fluid, natural animal-plant product material liquid can be used for numerous areas such as biochemical separation.
3. native system is convenient to operation, is easy to control, can realize sample on column buffer solution balance related in the integral post chromatography process, the material liquid, the flushing of post bed, eluent wash-out and regenerated liquid automatically to the operating procedures such as regeneration of column, and the target organism material is collected automatically.
Description of drawings
Fig. 1 is the hardware connection diagram of the automatic chromatography of integral post of the present invention system.
The specific embodiment
Below in conjunction with specific embodiment the automatic chromatography of integral post provided by the invention system is described in further details.
As shown in fig. 1, native system is made up of mechanical part and control section, mechanical part comprise at least sample introduction with wriggling pumping module, whole column, detector, go out sample and product collection module, control section is core with the single-chip microcomputer, comprise input-output unit, two above relays and three above drivers.
Sample introduction comprises four above sample control valves with wriggling pumping module, three above peristaltic pumps, six above fluid reservoirs and blender, the fluid reservoir that wherein is used for each eluent of storage is communicated with blender by the corresponding peristaltic pump that is used for this eluent of pumping respectively, blender is communicated with the fluid reservoir A that is used for storage mixing eluent, by being used for carrying the peristaltic pump A of liquid to comprise fluid reservoir A at least with the fluid reservoir that whole column injection port is communicated with to whole column, the fluid reservoir B that is used for storage of raw materials liquid, the fluid reservoir D that is used for storing the fluid reservoir C of equilibrium liquid and is used for store regenerated liquid is at fluid reservoir A, fluid reservoir B, be provided with sample control valve separately between fluid reservoir C and fluid reservoir D and the peristaltic pump A.
Go out that sample comprises with the product collection module going out the sample control valve more than three, be used for collecting object and the liquid collecting jar of waste liquid more than three, whole column outlet is communicated with the liquid collecting jar that respectively is used for collecting object and waste liquid, at whole column outlet and respectively be provided with out the sample control valve separately between the liquid collecting jar for collection object and waste liquid.
Detector is located at whole column outlet and is respectively gone out between the sample control valve.
Wherein, single-chip microcomputer is used for driving the driver of each peristaltic pump of control and controls sample introduction and wriggling pumping module for the relay A that drives sample control valve by driving, single-chip microcomputer is controlled out sample and product collection module by the relay B that drive to be used for driving the sample control valve, and output signal from detector is gathered and changed to single-chip microcomputer by the A/D conversion chip.
Single-chip microcomputer drives be used to the driver that drives each peristaltic pump of control respectively, is used for driving the relay A of each sample control valve with its drive circuit that carries, and is used for the relay B that driving respectively goes out the sample control valve.
In described control section, input-output unit comprises host computer, touch-screen, LCDs, host computer is by serial ports and single chip communication, touch-screen is connected with the I/O interface of single-chip microcomputer by touch-screen digital translation chip, and LCDs is connected with the I/O interface of single-chip microcomputer by the liquid crystal display driver.
Used detector is Ultraviolet Detector.
Also be provided with sensing module in the mechanical part, described sensing module comprises temperature sensor at least, two above liquid-pressure pick-ups, two above electrical conductivity unit and two above pH test cells, wherein be communicated with blender also and single chip communication by pipeline for detection of the electrical conductivity unit that mixes the eluent electrical conductivity and for detection of the pH test cell that mixes eluent pH value, electrical conductivity unit for detection of liquid electric conductivity in the integral post, for detection of the liquid-pressure pick-up of the fluid pressure of carrying to integral post and for detection of the temperature sensor of fluid temperature in the whole column is communicated with the outlet of peristaltic pump A by pipeline and and single chip communication, the pH test cell that reaches for detection of whole column liquid pH value for detection of the liquid-pressure pick-up of whole column fluid pressure is communicated with the outlet of whole column also and single chip communication by pipeline.
Described sensing module also comprises the air pressure probe for detection of the air capacity in the system.
System provided by the invention at first sets technological process and the relevant parameter of native system by touch-screen or host computer when work.Then, single-chip microcomputer is according to the flow process and the parameter that set, drive relay A with the switching of controlling each sample control valve and drive start and stop and the rotating speed of each driver to control each corresponding peristaltic pump by self drive circuit simultaneously by self drive circuit, regulate the flow of fluid and provide the required pressure of fluid each eluent is pumped into the mixing that realizes gradient eluent in the blender for whole tomographic system, and make material liquid, equilibrium liquid, eluent and regenerated liquid pass through whole column through sample control valve and peristaltic pump A successively according to the preset program of single-chip microcomputer, thereby realize related column buffer solution balance in the integral post chromatography process, sample on the material liquid, the flushing of post bed, eluent wash-out and regenerated liquid are to the steps such as regeneration of column.With Ultraviolet Detector the liquid that flows out from whole column is detected, the detection signal from detector is gathered and changed to single-chip microcomputer, testing result is analyzed, controlled out the switch of sample control valve according to this result, thereby can realize the categorised collection of each object and waste liquid.Simultaneously, can utilize temperature sensor, liquid-pressure pick-up, electrical conductivity unit and pH test cell in the sensing module to come parameters such as the material pressure in the chromatography process, temperature, electrical conductivity, pH are carried out detecting in real time automatically, thereby realize the real-time monitoring to whole technical process.
Because the macroporous type continuous bed complicated feed liquid that allows to contain microbial cell is directly passed through, thereby system provided by the present invention can realize containing the direct high speed full automatic treatment of the complicated feed liquid of microbial cell or cell fragment.
The separating medium aperture of native system can reach several microns even hundreds of micron, can allow to contain the microbial cell of submicron-scale or the complicated material liquid of cell fragment directly passes bed, can be used for directly separate targets biomolecular material from the complicated material liquid that contains solid phase impurity such as microbial cell or cell fragment.
The unit interval feed liquid treating capacity of native system is big, separates rapidly, and directly quickly separating target biomolecular material from complicated nutrient solution, zymotic fluid, natural animal-plant product material liquid can be used for numerous areas such as biochemical separation.
Native system is convenient to operation, is easy to control, can realize sample on column buffer solution balance related in the integral post chromatography process, the material liquid, the flushing of post bed, eluent wash-out and regenerated liquid automatically to the operating procedures such as regeneration of column, and the target organism material is collected automatically.
Claims (6)
1. integral post chromatography automatic separation system, it is characterized in that: formed by mechanical part and control section, mechanical part comprise at least sample introduction with wriggling pumping module, whole column, detector, go out sample and product collection module, control section is core with the single-chip microcomputer, comprise input-output unit, two above relays and three above drivers;
Sample introduction comprises four above sample control valves with wriggling pumping module, three above peristaltic pumps, six above fluid reservoirs, and blender, the fluid reservoir that wherein is used for each eluent of storage is first fluid reservoir, be communicated with blender by the corresponding peristaltic pump that is used for this eluent of pumping respectively, blender is communicated with the fluid reservoir A that is used for storage mixing eluent, peristaltic pump A is used for carrying liquid to whole column, the fluid reservoir that is communicated with whole column injection port by peristaltic pump A is second fluid reservoir, at least the second fluid reservoir comprises fluid reservoir A, the fluid reservoir B that is used for storage of raw materials liquid, the fluid reservoir D that is used for storing the fluid reservoir C of equilibrium liquid and is used for store regenerated liquid is at fluid reservoir A, fluid reservoir B, be provided with sample control valve separately between fluid reservoir C and fluid reservoir D and the peristaltic pump A;
Go out that sample comprises with the product collection module going out the sample control valve more than three, be used for collecting object and the liquid collecting jar of waste liquid more than three, whole column outlet is communicated with the liquid collecting jar that respectively is used for collecting object and waste liquid, at whole column outlet and respectively be provided with out the sample control valve separately between the liquid collecting jar for collection object and waste liquid;
Detector is located at whole column outlet and is respectively gone out between the sample control valve;
Wherein, single-chip microcomputer is used for driving the driver of each peristaltic pump of control and controls sample introduction and wriggling pumping module for the relay A that drives sample control valve by driving, single-chip microcomputer is controlled out sample and product collection module by the relay B that drive to be used for driving the sample control valve, and output signal from detector is gathered and changed to single-chip microcomputer by the A/D conversion chip.
2. integral post chromatography automatic separation system according to claim 1, it is characterized in that: single-chip microcomputer drives be used to the driver that drives each peristaltic pump of control respectively, is used for driving the relay A of each sample control valve with its drive circuit that carries, and is used for the relay B that driving respectively goes out the sample control valve.
3. integral post chromatography automatic separation system according to claim 1, it is characterized in that: in described control section, input-output unit comprises host computer, touch-screen, LCDs, host computer is by serial ports and single chip communication, touch-screen is connected with the I/O interface of single-chip microcomputer by touch-screen digital translation chip, and LCDs is connected with the I/O interface of single-chip microcomputer by the liquid crystal display driver.
4. integral post chromatography automatic separation system according to claim 1, it is characterized in that: used detector is Ultraviolet Detector.
5. integral post chromatography automatic separation system according to claim 1, it is characterized in that: also be provided with sensing module in the mechanical part, described sensing module comprises temperature sensor at least, two above liquid-pressure pick-ups, two above electrical conductivity unit and two above pH test cells, wherein be communicated with blender also and single chip communication by pipeline for detection of the electrical conductivity unit that mixes the eluent electrical conductivity and for detection of the pH test cell that mixes eluent pH value, electrical conductivity unit for detection of liquid electric conductivity in the integral post, for detection of the liquid-pressure pick-up of the fluid pressure of carrying to integral post and for detection of the temperature sensor of fluid temperature in the whole column is communicated with the outlet of peristaltic pump A by pipeline and and single chip communication, the pH test cell that reaches for detection of whole column liquid pH value for detection of the liquid-pressure pick-up of whole column fluid pressure is communicated with the outlet of whole column also and single chip communication by pipeline.
6. integral post chromatography automatic separation system according to claim 5, it is characterized in that: described sensing module also comprises the air pressure probe for detection of the air capacity in the system.
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| CN 201110129336 CN102258885B (en) | 2011-05-18 | 2011-05-18 | Automatic separation system for integrated column chromatography |
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| CN 201110129336 CN102258885B (en) | 2011-05-18 | 2011-05-18 | Automatic separation system for integrated column chromatography |
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| CN102258885B true CN102258885B (en) | 2013-08-21 |
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| CN102527089B (en) * | 2012-02-01 | 2013-10-30 | 中国矿业大学 | Multi-segment chromatographic column |
| CN103698289A (en) * | 2013-12-05 | 2014-04-02 | 成都雅途生物技术有限公司 | Chromatographic column effluent detection device |
| CN107422664A (en) * | 2017-06-07 | 2017-12-01 | 利穗科技(苏州)有限公司 | UV, PH based on protein chromatography system, conductance comprehensive detection system |
| CN108019341A (en) * | 2017-12-04 | 2018-05-11 | 苏州市海崴生物科技有限公司 | A kind of basic model quaternary diaphragm pump |
| CN107992007A (en) * | 2017-12-04 | 2018-05-04 | 苏州市海崴生物科技有限公司 | One kind digitlization tomographic system |
| CN108279625B (en) * | 2017-12-30 | 2023-09-05 | 利穗科技(苏州)有限公司 | Digital chromatography system and method |
| CN109453545A (en) * | 2018-12-20 | 2019-03-12 | 上海兆维科技发展有限公司 | The liquid dispensing device of Column eluate |
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| US6635173B2 (en) * | 2000-12-28 | 2003-10-21 | Cohesive Technologies, Inc. | Multi column chromatography system |
| CN1346984A (en) * | 2001-11-07 | 2002-05-01 | 湖南力合科技发展有限公司 | High-speed automatic colorimetric analyzer |
| CN201161989Y (en) * | 2007-08-14 | 2008-12-10 | 百奇生物科技(上海)有限公司 | Multicenter protein compatible chromatography purification instrument |
| CN101664571B (en) * | 2008-09-05 | 2012-02-01 | 中国人民解放军第三军医大学第二附属医院 | Device for treating dysfunction of central nervous system |
| CN201280551Y (en) * | 2008-10-20 | 2009-07-29 | 中国农业大学 | System for purifying bacteria magnetosome |
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