CN102258885B - Automatic separation system for integrated column chromatography - Google Patents

Abstract

The invention discloses an integral column chromatography automatic separation system, which is composed of a mechanical part and a control part. The mechanical part at least includes a sample injection and peristaltic pumping module, an integral bed column, a detector, a sample discharge and product collection module, and the control part includes Single-chip microcomputer, input and output unit, relay and driver; the integral bed column sample inlet is connected with the peristaltic pump A in the sample injection and peristaltic pumping module, and its sample outlet is connected with the liquid collection tank in the sample output and product collection module, and the detector is set Between the integral bed column sample outlet and the sample and product collection module; the single-chip microcomputer controls the sample injection and peristaltic pumping module and the sample and product collection module, and collects and converts the signal from the detector through the A/D conversion chip. The system provided by the invention allows complex feed liquids containing microbial cells to pass directly, and can directly and automatically process complex feed liquids containing microbial cells or cell fragments at high speed.

Description

Monolithic column chromatography automatic separation system

technical field

The invention relates to a monolithic column automatic chromatographic separation system, which belongs to the technical fields of biology, chemical industry and machinery.

Background technique

Macroporous monolithic column chromatography (also known as monolithic bed or continuous bed chromatography) is a new type of biological separation method developed in recent years. The pore size of the monolithic column separation medium can reach several microns or even hundreds of microns, allowing sub- The complex raw material solution of micron-scale microbial cells or cell fragments passes directly through the bed, and the adsorption chromatography separation of the target in this type of separation medium is rapid, which can be used to directly extract the complex liquid containing solid phase impurities such as microbial cells or cell fragments. The target biomolecular substances are separated from the raw material solution. This method has the comprehensive advantages of expanded bed and conventional fixed bed chromatography. It can integrate several steps of filtration, centrifugation, concentration and chromatographic separation in one step. The processing capacity per unit time is large, and it is suitable for the rapid preparation of biochemical substances. separate. Therefore, it has broad application prospects in the separation and purification of microbial fermentation biochemicals, natural product extraction or genetic engineering drugs.

Although there are many automatic separation systems for biological chromatography at home and abroad, such as expanded bed chromatography system, conventional medium and low pressure fixed bed chromatography system and high performance liquid chromatography system, but in addition to the expanded bed system, the existing Most of the chromatographic systems are aimed at analytical separation or small-scale preparation separation, and complex feed-liquid fluids containing solid-phase substances such as microbial cells or cell fragments are not allowed to pass through the bed column, so it is difficult to directly perform rapid separation of complex fluids. At present, there is no special automatic chromatography system for macroporous monolithic column separation at home and abroad. The existing high performance liquid chromatography system or assembled chromatography system is inconvenient to operate, and it is not applicable or has certain difficulties in large-scale preparation and separation.

Contents of the invention

In order to solve the above-mentioned problems in the prior art, the present invention provides an integral column chromatography automatic separation system. The system of the present invention allows complex feed liquids containing microbial cells to pass directly, and can directly and automatically process liquids containing microbial cells or cell fragments at high speed. complex liquid.

In order to achieve the above purpose, the present invention adopts the following technical solutions: an automatic separation system for integral column chromatography, which is composed of a mechanical part and a control part. The mechanical part at least includes a sample injection and peristaltic pumping module, an integral bed column, a detector, a With the product collection module, the control part is based on the single-chip microcomputer, including input and output units, more than two relays and more than three drivers;

The sample injection and peristaltic pumping module includes more than four sample injection control valves, more than three peristaltic pumps, more than six liquid storage tanks and mixers, in which the liquid storage tanks used to store eluents pass through corresponding The peristaltic pump for pumping the eluent communicates with the mixer, the mixer communicates with the liquid storage tank A for storing the mixed eluent, and communicates with the monolithic bed column inlet through the peristaltic pump A for delivering liquid to the monolithic bed column The liquid storage tank at least includes liquid storage tank A, liquid storage tank B for storing raw material liquid, liquid storage tank C for storing balance liquid and liquid storage tank D for storing regeneration liquid, in liquid storage tank A, There are sampling control valves between the liquid storage tank B, the liquid storage tank C and the liquid storage tank D and the peristaltic pump A respectively;

The sample and product collection module includes more than three sample control valves, more than three liquid collection tanks for collecting target objects and waste liquids, integral bed column sample outlets and liquid collection tanks for collecting target objects and waste liquids Connected, a sample control valve is provided between the integral bed column sample outlet and each liquid collection tank used to collect the target substance and waste liquid;

The detector is set between the integral bed column sample outlet and each sample control valve;

Among them, the single-chip microcomputer controls the sampling and peristaltic pumping module by driving the driver for controlling each peristaltic pump and the relay A for driving the sampling control valve, and the single-chip microcomputer controls it by driving the relay B for driving the sampling control valve. In the sample and product collection module, the single-chip microcomputer collects and converts the output signal from the detector through the A/D conversion chip.

The single-chip microcomputer uses its own drive circuit to respectively drive the driver for controlling each peristaltic pump, the relay A for driving each sample injection control valve, and the relay B for driving each sample output control valve.

In the control part, the input and output unit includes a host computer, a touch screen, and a liquid crystal display. The host computer communicates with the single-chip computer through a serial port, and the touch screen is connected to the I/O interface of the single-chip microcomputer through a touch screen digital conversion chip. Connect with the I/O interface of the one-chip computer.

The detector used is a UV detector.

There is also a sensor module in the mechanical part, and the sensor module includes at least a temperature sensor, more than two liquid pressure sensors, more than two conductivity units and more than two pH test units, which are used to detect the conductivity of the mixed eluent The conductivity unit for detecting the pH value of the mixed eluent and the pH testing unit for detecting the pH value of the mixed eluent communicate with the mixer through a pipeline and communicate with the single-chip microcomputer. The liquid pressure sensor for the pressure of the liquid being delivered and the temperature sensor for detecting the temperature of the liquid in the integral bed column communicate with the outlet of the peristaltic pump A through a pipeline and communicate with the single-chip microcomputer, and the liquid pressure sensor for detecting the liquid pressure of the integral bed column and for detecting The pH test unit of the liquid pH value of the integral bed column communicates with the sample outlet of the integral bed column through a pipeline and communicates with the single-chip computer.

The sensing module also includes an air pressure sensor for detecting the amount of air in the system.

Compared with the prior art, the integral column chromatography automatic separation system provided by the present invention has the following advantages:

1. The pore size of the separation medium of this system can reach several microns or even hundreds of microns, which can allow the complex raw material solution containing submicron microbial cells or cell fragments to pass directly through the bed layer, and can be used to directly separate the liquid containing microbial cells or cell fragments, etc. Separation of target biomolecular substances from complex feedstock solutions with solid phase impurities.

2. The system has a large amount of feed liquid per unit time and rapid separation. It can directly and quickly separate target biomolecular substances from complex culture liquids, fermentation liquids, and natural animal and plant product raw material liquids. It can be used in many fields such as biochemical separation.

3. The system is easy to operate and control, and can automatically realize the bed column buffer balance, raw material liquid loading, column bed flushing, eluent elution and regeneration of the bed column involved in the whole column chromatography separation process, etc. Operation steps, and automatic collection of target biological substances.

Description of drawings

Fig. 1 is a schematic diagram of the hardware connection of the monolithic column automatic chromatographic separation system of the present invention.

Detailed ways

The monolithic column automatic chromatographic separation system provided by the present invention will be further described in detail below in conjunction with specific examples.

As shown in Figure 1, the system consists of a mechanical part and a control part. The mechanical part at least includes a sample injection and peristaltic pumping module, an integral bed column, a detector, a sample output and a product collection module. The control part is centered on a single-chip microcomputer, including Input and output units, more than two relays and more than three drivers.

The sample injection and peristaltic pumping module includes more than four sample injection control valves, more than three peristaltic pumps, more than six liquid storage tanks and mixers, in which the liquid storage tanks used to store eluents pass through corresponding The peristaltic pump for pumping the eluent communicates with the mixer, the mixer communicates with the liquid storage tank A for storing the mixed eluent, and communicates with the monolithic bed column inlet through the peristaltic pump A for delivering liquid to the monolithic bed column The liquid storage tank at least includes liquid storage tank A, liquid storage tank B for storing raw material liquid, liquid storage tank C for storing balance liquid and liquid storage tank D for storing regeneration liquid, in liquid storage tank A, A sampling control valve is provided between the liquid storage tank B, the liquid storage tank C and the liquid storage tank D and the peristaltic pump A respectively.

The sample and product collection module includes more than three sample control valves, more than three liquid collection tanks for collecting target objects and waste liquids, integral bed column sample outlets and liquid collection tanks for collecting target objects and waste liquids Connected, and a sample control valve is provided between the integral bed column sample outlet and each liquid collection tank used to collect the target substance and waste liquid.

The detector is set between the sample outlet of the integral bed column and each sample control valve.

Among them, the single-chip microcomputer controls the sampling and peristaltic pumping module by driving the driver for controlling each peristaltic pump and the relay A for driving the sampling control valve, and the single-chip microcomputer controls it by driving the relay B for driving the sampling control valve. In the sample and product collection module, the single-chip microcomputer collects and converts the output signal from the detector through the A/D conversion chip.

The single-chip microcomputer uses its own drive circuit to respectively drive the driver for controlling each peristaltic pump, the relay A for driving each sample injection control valve, and the relay B for driving each sample output control valve.

In the control part, the input and output unit includes a host computer, a touch screen, and a liquid crystal display. The host computer communicates with the single-chip computer through a serial port, and the touch screen is connected to the I/O interface of the single-chip microcomputer through a touch screen digital conversion chip. Connect with the I/O interface of the one-chip computer.

The detector used is a UV detector.

There is also a sensor module in the mechanical part, and the sensor module includes at least a temperature sensor, more than two liquid pressure sensors, more than two conductivity units and more than two pH test units, which are used to detect the conductivity of the mixed eluent The conductivity unit for detecting the pH value of the mixed eluent and the pH testing unit for detecting the pH value of the mixed eluent communicate with the mixer through a pipeline and communicate with the single-chip microcomputer. The liquid pressure sensor for the pressure of the liquid being delivered and the temperature sensor for detecting the temperature of the liquid in the integral bed column communicate with the outlet of the peristaltic pump A through a pipeline and communicate with the single-chip microcomputer, and the liquid pressure sensor for detecting the liquid pressure of the integral bed column and for detecting The pH test unit of the liquid pH value of the integral bed column communicates with the sample outlet of the integral bed column through a pipeline and communicates with the single-chip computer.

The sensing module also includes an air pressure sensor for detecting the amount of air in the system.

When the system provided by the present invention is working, firstly, the process flow and related parameters of the system are set through the touch screen or the host computer. Then, according to the set process and parameters, the single-chip microcomputer drives the relay A through its own driving circuit to control the opening and closing of each sampling control valve, and at the same time drives each driver through its own driving circuit to control the start-up of each corresponding peristaltic pump. Stop and rotate, adjust the flow of the fluid and provide the pressure required by the fluid for the entire chromatography system to pump each eluent into the mixer to achieve the mixing of the gradient eluent, and make the raw material liquid, balance liquid, and elution liquid The solution and the regeneration solution pass through the monolithic bed column sequentially through the injection control valve and the peristaltic pump A in accordance with the predetermined program of the single-chip microcomputer, so as to realize the balance of the bed column buffer solution, loading of the raw material solution, column bed washing, washing and so on involved in the monolithic column chromatography separation process. Steps such as dehydration, elution and regeneration of the bed column by the regeneration solution. Use an ultraviolet detector to detect the liquid flowing out of the integral bed column, the single-chip microcomputer collects and converts the detection signal from the detector, analyzes the detection result, and controls the switch of the sample control valve according to the result, so as to realize the target and Sorted collection of waste liquid. At the same time, the temperature sensor, liquid pressure sensor, conductivity unit and pH test unit in the sensing module can be used to automatically detect the material pressure, temperature, conductivity, pH and other parameters in the chromatography separation process in real time, so as to realize the Real-time monitoring of the entire process.

Since the macroporous continuous bed allows the complex feed liquid containing microbial cells to pass directly, the system provided by the present invention can realize the direct high-speed automatic processing of the complex feed liquid containing microbial cells or cell fragments.

The pore size of the separation medium in this system can reach several microns or even hundreds of microns, which can allow the complex raw material liquid containing submicron microbial cells or cell fragments to pass directly through the bed, and can be used to directly separate from solid phases containing microbial cells or cell fragments. Separation of target biomolecular substances from complex feedstock solutions with impurities.

The system has a large amount of feed liquid per unit time and rapid separation. It can directly and quickly separate target biomolecular substances from complex culture liquids, fermentation liquids, and natural animal and plant product raw material liquids, and can be used in many fields such as biochemical separation.

The system is easy to operate and control, and can automatically realize the operation steps involved in the whole column chromatography separation process, such as bed column buffer balance, raw material solution loading, column bed flushing, eluent elution, and regeneration solution for bed column regeneration. , and automatically collect target biological substances.

Claims (6)

1. An integral column chromatography automatic separation system, characterized in that: it is composed of a mechanical part and a control part, the mechanical part at least includes a sample injection and peristaltic pumping module, an integral bed column, a detector, a sample and a product collection module, and the control Part of them is based on single-chip microcomputer, including input and output units, more than two relays and more than three drivers; The sampling and peristaltic pumping module includes more than four sampling control valves, more than three peristaltic pumps, more than six liquid storage tanks, and a mixer, wherein the liquid storage tanks used to store each eluent are the first liquid storage The tanks communicate with the mixer through the corresponding peristaltic pumps used to pump the eluent, the mixer communicates with the liquid storage tank A for storing the mixed eluent, and the peristaltic pump A is used to deliver the liquid to the monolithic bed column, The liquid storage tank communicated with the integral bed column sampling port through the peristaltic pump A is the second liquid storage tank, at least the second liquid storage tank includes the liquid storage tank A, the liquid storage tank B for storing the raw material liquid, and the liquid storage tank B for storing the balance liquid The liquid storage tank C and the liquid storage tank D used to store the regeneration liquid are respectively provided with sampling control valves between the liquid storage tank A, the liquid storage tank B, the liquid storage tank C and the liquid storage tank D and the peristaltic pump A ; The sample and product collection module includes more than three sample control valves, more than three liquid collection tanks for collecting target objects and waste liquids, integral bed column sample outlets and liquid collection tanks for collecting target objects and waste liquids Connected, a sample control valve is provided between the integral bed column sample outlet and each liquid collection tank used to collect the target substance and waste liquid; The detector is set between the integral bed column sample outlet and each sample control valve; Among them, the single-chip microcomputer controls the sampling and peristaltic pumping module by driving the driver for controlling each peristaltic pump and the relay A for driving the sampling control valve, and the single-chip microcomputer controls it by driving the relay B for driving the sampling control valve. In the sample and product collection module, the single-chip microcomputer collects and converts the output signal from the detector through the A/D conversion chip. 2. The integral column chromatography automatic separation system according to claim 1, characterized in that: the single-chip microcomputer drives respectively the driver for driving and controlling each peristaltic pump, the driver for driving each sample injection control valve with its own driving circuit. Relay A, and relay B for driving each sample control valve. 3. The integral column chromatography automatic separation system according to claim 1, characterized in that: in the control part, the input and output unit includes a host computer, a touch screen, and a liquid crystal display, and the host computer communicates with the single-chip microcomputer through a serial port, and the touch screen The digital conversion chip of the touch screen is connected with the I/O interface of the single-chip microcomputer, and the liquid crystal display is connected with the I/O interface of the single-chip microcomputer through a liquid crystal screen driver. 4. The integral column chromatography automatic separation system according to claim 1, characterized in that: the detector used is an ultraviolet detector. 5. The integral column chromatography automatic separation system according to claim 1, characterized in that: a sensor module is also provided in the mechanical part, and the sensor module at least includes a temperature sensor, more than two liquid pressure sensors, two The above conductivity unit and more than two pH test units, wherein the conductivity unit used to detect the conductivity of the mixed eluent and the pH test unit used to detect the pH value of the mixed eluent communicate with the mixer through a pipeline and communicate with the single-chip microcomputer , the conductivity unit used to detect the conductivity of the liquid in the monolithic column, the liquid pressure sensor used to detect the pressure of the liquid delivered to the monolithic column, and the temperature sensor used to detect the temperature of the liquid in the monolithic bed column pass through the pipeline and the outlet of the peristaltic pump A The liquid pressure sensor used to detect the liquid pressure of the integral bed column and the pH test unit used to detect the pH value of the liquid of the integral bed column communicate with the sample outlet of the integral bed column through pipelines and communicate with the single-chip microcomputer. 6. The integral column chromatography automatic separation system according to claim 5, characterized in that: the sensing module further comprises an air pressure sensor for detecting the air volume in the system.

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