CN101164700A - Continuous production equipment for biochemical product magnetic adsorption and desorption - Google Patents

Abstract

The invention belongs to the technical field of adsorption and separation of biochemical products, in particular to continuous production equipment for magnetic adsorption and desorption of biochemical products. In the present invention, the air-lift adsorption tower (tower 1), the air-lift desorption tower (tower 2) and the storage tank are combined, and distribution plates are arranged at the bottom of the two towers, and the microspheres and the raw material liquid are realized by means of bubble dispersion and stirring. The thorough mixing of tower 1 and tower 2 and tower 2 and the storage tank are connected with a very thin square tube for transporting magnetic microspheres, and a return line is provided between the storage tank and tower 1 for magnetic microspheres recycle and re-use. The magnetic separation equipment connected at the outlet of the two towers is composed of a track-shaped magnet group, and the magnet group is driven by a motor to rotate at a certain speed, so that the magnetic microspheres can be enriched and separated under the action of the rotating magnet .

Description

Continuous production equipment for magnetic adsorption and desorption of biochemical products

technical field

The invention belongs to the technical field of adsorption and separation of biochemical products, in particular to continuous production equipment for magnetic adsorption and desorption of biochemical products.

technical background

Superparamagnetic microspheres have the advantages that ordinary solid particles do not have: first, it has superparamagnetism and exhibits good magnetism under the condition of an external magnetic field. When the external magnetic field is removed, the remanence of the magnetic microsphere carrier is zero. This property can be conveniently located, separated, and recovered; secondly, the surface of the magnetic microspheres can be grafted and modified according to needs, and different functional groups can be attached to the magnetic microspheres to give them different bioselectivities, which in turn can Separation of biological products such as natural products, proteins, nucleic acids, and cells; in addition, the magnetic microspheres have the advantages of small particles, large specific surface area, and low dosage. When performing operations such as adsorption, desorption, or reaction separation, the unit operation speed is greatly accelerated. At the same time, it also reduces the consumption of raw materials, thus improving production efficiency.

The traditional adsorption separation of biochemical products is usually divided into two independent unit operations of adsorption (reaction) and desorption (separation). For the adsorption (reaction) unit operation, in the past, mechanical stirring or electromagnetic stirring was used in the reactor to make magnetic particles adsorb or perform physical and chemical reactions in the reactor. Now, in order to prevent the tangential stirring force of mechanical stirring from breaking microorganisms Cells or biochemical products are destroyed, and the method of bubbling with _O2 and other gases is used instead of mechanical stirring to achieve production applications. For the unit operation of desorption (separation), the product is obtained, and the two unit operations of adsorption and desorption require the separation of magnetic microspheres. The traditional process often uses intermittent operations such as filtration, which has low production efficiency and troublesome operation. .

The existing magnetic separation technology and equipment usually use electromagnetic field, which has weak magnetic field strength and high equipment cost. The permanent magnet is now used as the main part of the magnetic separation equipment, which can not only reduce energy consumption and production cost, but also increase the magnetic field strength and improve production efficiency.

There are many reports about magnetic reaction devices and magnetic separation devices, such as patent CN: 86104692, which involves only a magnetic drum separator, and does not integrate reaction and separation or adsorption and desorption, such as patent CN: 02120810 The design is an integrated device of reaction and separation, but the dispersion effect of the reactor part in the device is not obvious. The present invention not only integrates the operation of reaction and separation or adsorption and desorption, but also adopts the method of bubbling in the biochemical reactor to perform tower Dispersion within. The invention integrates the bioreactor dispersion technology, magnetic separation technology and unit operation integration technology, which is conducive to continuous operation, scale expansion, cost reduction and product yield improvement, and is one of the important directions of chemical industry development at present.

Contents of the invention

The purpose of the invention is to provide continuous production equipment for magnetic adsorption and desorption of biochemical products.

The continuous production equipment of biochemical product magnetic adsorption and desorption of the present invention as shown in Figure 1, comprises air-lift type adsorption (reaction) tower (tower 1), air-lift type desorption (separation) tower (tower 2), distribution plate, magnet groups, storage tanks, etc.;

An air-lift adsorption (reaction) tower has an inlet and an air inlet for the raw material liquid at the bottom of the tower, and a distribution plate for the dispersion and mixing of the raw material liquid and air bubbles is installed in the lower part of the tower. There is an inlet for magnetic microspheres above the distribution plate, and a pipeline connected to the storage tank is installed at the inlet for recycling of magnetic microspheres; an air outlet is opened on the top of the tower for balancing tank pressure There is a feed liquid outlet on the side of the upper part of the tower, a pipeline with two outlets is connected at the outlet, a magnet group connected to the ring-shaped crawler is installed at the pipeline, and a magnet group connected to the ring-shaped crawler is installed at the pipeline. Driven by a motor to rotate, one outlet of the pipeline is used to lead out the waste liquid, and the other outlet is connected to a pipeline connected with the air-lift desorption (separation) tower (tower 2) (the magnetic microspheres that have adsorbed the product are introduced into the lower a tower);

An air-lift desorption (separation) tower, with an eluent inlet and air inlet at the bottom of the tower, a distribution plate for eluent and air bubble dispersion and mixing is installed in the lower part of the tower, and at the lower side of the tower, And above the distribution plate, there is a pipeline connected with the air-lift adsorption (reaction) tower (the magnetic microspheres that have adsorbed the product are introduced from this inlet); there is a gas outlet on the top of the tower to balance the tank pressure. There is a feed liquid outlet on the side of the upper part of the tower, a pipeline with two outlets is connected at the outlet, a magnet group connected to the ring-shaped crawler is installed at the pipeline, and a magnet group connected to the ring-shaped crawler is installed at the pipeline. Driven by the motor to rotate, the outlet of one pipeline is used to export the concentrated product, and the other outlet is connected to a pipeline connected with the storage tank (for magnetic microsphere recovery), and the magnetic microspheres desorbed from the product are introduced;

A pipeline connects the storage tank with the air-lift adsorption (reaction) tower, and under the action of compressed air, the recovered magnetic microspheres are introduced into the lower part of the air-lift adsorption (reaction) tower.

The capacity of described air-lift desorption (separation) tower is much smaller than air-lift adsorption (reaction) tower, also can find out by schematic diagram 1, this is because the adsorption of product in the feed liquid is carried out in tower 1, tower 2 The middle stage is to carry out the desorption of the product on the magnetic microspheres, which is a product concentration process relative to tower 1.

The magnets that are connected on the track are permanent magnets, and the N or S poles of all magnets are towards the outside of the track; the direction of rotation of the track is that the magnetic microspheres flow out of the air-lift adsorption (reaction) tower or the air-lift desorption ( separation) tower direction. The magnet group deposits the magnetic microspheres in the pipeline, and flows into the pipeline next to the magnet group connecting the air-lift adsorption (reaction) tower and the air-lift desorption (separation) tower, and connects the air-lift Desorption (separation) tower and the pipeline of the storage tank are enriched.

The tower, storage tank and pipeline are made of stainless steel, glass or plastic.

In the present invention, as shown in Figure 1, the raw material liquid and gas are introduced from the bottom of tower 1 together, and after being dispersed by the distribution plate, they are fully mixed with the magnetic microspheres that flow in from the side of the tower bottom through desorption and recovery activity in the tower, and the gas in the raw material liquid is fully mixed. The product that needs to be recovered is adsorbed (reacted), and the magnetic microspheres that have adsorbed the product are coagulated under the magnetic force of the rotating magnet group, and are introduced into the bottom of the tower 2 by a thin tube below the rotating magnet, and the raw material solution ( Waste liquid) is discharged from a thick pipe above. After the magnetic microspheres that have adsorbed the product enter tower 2, they are fully mixed with the eluent under the action of air bubbles and interact with each other for elution. The magnetic microspheres of the product are collected by a thin tube under the action of the rotating magnet group, enriched and recovered to the storage tank, and transported to the bottom of tower 1 under the action of compressed air for recycling.

In the present invention, the air-lift adsorption tower, the air-lift desorption tower and the storage tank are combined together, distribution plates are placed at the bottom of the two towers, and the microspheres and the raw material liquid are fully mixed by means of bubble dispersion and stirring. Tower 1 and tower 2 and the tower 2 and the storage tank are connected with a very thin square tube for transporting the magnetic microspheres, and a return pipeline is provided between the storage tank and the tower 1 for recycling the magnetic microspheres. The magnetic separation equipment connected at the outlet of the two towers is composed of a crawler-shaped magnet group, and the magnet group is driven by a motor to rotate at a certain speed, so that the magnetic microspheres can be enriched and separated under the action of the rotating magnet .

Description of drawings

Figure 1. Schematic representation of the apparatus of the present invention.

Fig. 2. the magnetic carrier that the experiment of embodiment 1 of the present invention obtains is to bovine serum albumin (BSA) adsorption amount and desorption amount (under 1M NaCl condition) change curve with time.

reference sign

1, 4. Distribution plate 2. Raw material liquid level 3. Eluent liquid level

5, 6. Magnet group 7. Storage tank

8. Magnetic microspheres that have adsorbed products 9. Magnetic microspheres that have desorbed products

Detailed ways

Example 1.

See Figure 1. An air-lift adsorption tower (tower 1), with an inlet and an air inlet for the raw material liquid at the bottom of the tower, a distribution plate 1 for the dispersion and mixing of the raw material liquid 2 and air bubbles is installed in the lower part of the tower, and the lower part of the side of the tower , and there is an inlet of magnetic microspheres above the distribution plate, and a pipeline connected with the storage tank is installed at the inlet for the recycling of magnetic microspheres; an outlet is opened at the top of the tower for Balance the tank pressure; there is a feed liquid outlet on the side of the upper part of the tower, a pipeline with two outlets is connected to the outlet, and a magnet group 5 connected to the ring-shaped track is installed on the pipeline, which is connected to the ring-shaped track. The magnet group on the top is driven by a motor to rotate, and the outlet of a pipeline with a valve is used to lead out the waste liquid, and the other outlet is connected to a pipeline connected with the air-lift desorption (separation) tower (tower 2). The magnetic microspheres 8 that have adsorbed the product are introduced into the next tower, i.e. the air-lift desorption (separation) tower;

An air-lift desorption (separation) tower (tower 2), has the inlet and air inlet of eluent at the bottom of the tower, and the distribution plate 4 that is used for eluent 3 and bubble dispersion mixing is housed in the lower part of the tower inside, At the lower side of the tower, and above the distribution plate, a pipeline communicating with the airlift adsorption (reaction) tower is installed (the magnetic microspheres adsorbed by the product are introduced from this inlet); there is a gas outlet at the top of the tower , used to balance the tank pressure; there is a feed liquid outlet on the side of the upper part of the tower, a pipeline with two outlets is connected at the outlet, and a magnet group 6 connected to the ring track is installed at the pipeline. The magnet group on the ring-shaped crawler is driven by a motor to rotate, and the outlet of a pipeline with a valve is used to export the concentrated product, and the other outlet is connected to a pipeline connected with the storage tank 7 (for magnetic microsphere recovery) , introducing the magnetic microspheres 9 that have desorbed the product;

A pipeline connects the storage tank with the air-lift adsorption (reaction) tower, and under the action of compressed air, the recovered magnetic microspheres are introduced into the lower part of the air-lift adsorption (reaction) tower.

1. The magnetic microspheres are prepared by grafting reactive triazine dyes on the surface of magnetic poly(ethylene glycol methacrylate) microspheres (PMMA-PEG-CB). Experiments have proved that the magnetic microspheres have superparamagnetism.

2. Experimental method of affinity adsorption and desorption: use poly(ethylene glycol methacrylate) reactive triazine dye magnetic microspheres (PMMA-PEG-CB) as a carrier to affinity-adsorb bovine serum albumin (BSA), and then The bovine serum albumin (BSA) on the carrier is desorbed under high salt concentration.

(1) Adsorption operation:

All operations of the magnetic microsphere affinity separation of bovine serum albumin (BSA) were carried out in Michaelis phosphate buffer solution with pH=6 and a concentration of 20 mM. Add 20ml of phosphate buffer containing 160mg of the magnetic microspheres into the stirring mixer, stir for several minutes, and add bovine serum albumin (BSA) concentration in addition and be 20ml of phosphate buffer of 2.0mg/ml to make bovine serum albumin (BSA ) total concentration is 1.0mg/ml, at the same time, start the precision metering infusion pump, control the solution flow at about 1ml/min, regularly sample and analyze the concentration of bovine serum albumin (BSA) at the outlet of the magnetic medium separation tube, and calculate the adsorption amount changes over time.

(2) Desorption operation:

The desorption kinetic curve of bovine serum albumin (BSA) on the magnetic microspheres was carried out in the phosphate buffer solution with a NaCl concentration of 1.0 M by the same measurement method.

3. The curves of the adsorption and desorption (under the condition of 1M NaCl) of the magnetic microspheres to bovine serum albumin (BSA) obtained by the experiment with time are shown in Figure 2.

Experimental conclusion: The superparamagnetic affinity separation microspheres have a high affinity adsorption capacity for bovine serum albumin (BSA) at low salt concentrations, and increasing the salt concentration can desorb most of bovine serum albumin (BSA) , Adsorption and desorption reached equilibrium within 10-20min.

Claims (5)

1. the continuous production equipment of a biochemical product magnetic adsorption and desorption comprises the gas-lifting type adsorption tower, gas-lifting type desorber, distribution grid, group of magnets, storage tank; It is characterized in that:

One gas-lifting type adsorption tower, have the import and the air inlet of material liquid in the bottom of tower, the distribution grid that is used for material liquid and bubble dispersion mixing is housed in the bottom of tower the inside, bottom, side at tower, and above distribution grid, have the inlet of magnetic microsphere, the pipeline that is connected with storage tank is installed in this porch; Top at tower has the gas outlet; Side on the top of tower has material liquid outlet, the pipeline that has two outlets in exit connection one, one group of magnets that is connected on the cast crawler belt is installed at the pipeline place, be connected group of magnets on the cast crawler belt by the motor driven rotary, the outlet of one pipeline is used to derive waste liquid, and another outlet connects a pipeline that is connected with the gas-lifting type desorber;

One gas-lifting type desorber, have the import and the air inlet of eluant, eluent in the bottom of tower, in the bottom of tower the inside the distribution grid that is used for eluant, eluent and bubble dispersion mixing is housed,, and the pipeline that is connected with the gas-lifting type adsorption tower is installed above distribution grid in the bottom, side of tower; Top at tower has the gas outlet; Side on the top of tower has material liquid outlet, the pipeline that has two outlets in exit connection one, one group of magnets that is connected on the cast crawler belt is installed at the pipeline place, be connected group of magnets on the cast crawler belt by the motor driven rotary, the outlet of one pipeline is used to derive the product after concentrating, another outlet connects a pipeline that is connected with storage tank, with desorb the magnetic microsphere importing of product;

One pipeline is connected storage tank with the gas-lifting type adsorption tower, the magnetic microsphere after will reclaiming under the effect of calming the anger imports the bottom of gas-lifting type adsorption tower.

2. equipment according to claim 1 is characterized in that: the described magnet that is connected on the crawler belt is permanent magnet.

3. equipment according to claim 1 and 2 is characterized in that: the N of the described whole magnet that are connected the magnet on the crawler belt or the S utmost point are outside crawler belt.

4. equipment according to claim 1 and 2 is characterized in that: described crawler belt direction of rotation is magnetic microsphere effluent gases lift-type adsorption tower or gas-lifting type desorber direction.

5. equipment according to claim 3 is characterized in that: described crawler belt direction of rotation is magnetic microsphere effluent gases lift-type adsorption tower or gas-lifting type desorber direction.

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