CN101972558A

CN101972558A - Expanded bed chromatographic separation column used for biochemical separation process and process flow

Info

Publication number: CN101972558A
Application number: CN 201010565313
Authority: CN
Inventors: 顾雄毅
Original assignee: Individual
Current assignee: Individual
Priority date: 2010-11-30
Filing date: 2010-11-30
Publication date: 2011-02-16
Anticipated expiration: 2030-11-30
Also published as: CN101972558B

Abstract

The invention relates to an expanded bed chromatographic separation column used for a biochemical separation process and a process flow, belonging to the field of biochemistry. In the invention, a liquid pressure equalizing distributor with self-cleaning function is arranged below a porous blocking sieve plate at the bottom of an expanded bed, and an upper nozzle assembly with back-flushing cleaning function is arranged at the top of the expanded bed so that the expanded bed chromatographic separation column is suitable for treating a raw material liquid containing solid particle impurities, meanwhile, the raw material sample liquid entering the expanded bed can be better distributed uniformly, therefore, the problem of uniform distribution of feed liquid pressure of lower-bed pressure drop feed liquid along a radius direction in the industrial amplification production process is solved, fluid passing through a bed layer of the expanded bed is better ensured to be in a plug flow state, the separation theoretical plate number and the chromatographic separation efficiency of the expanded bed can be greatly enhanced, the using effect and the using efficiency of the expanded bed are improved and the adsorption capacity and the desorption resolution of the expanded bed are enhanced.

Description

A kind of expanded bed chromatography splitter and technological process that is used for biochemical separation processes

Technical field

The invention belongs to technological field of biochemistry, relate in particular to and a kind ofly in industrial-scale production, be used to extract/chromatographic separation device and the production technology thereof of separation/purification.

Background technology

Chromatographic isolation is one of method the most frequently used in the biochemical separation processes.

Traditional chromatography column (abbreviation chromatographic column) adopts the fixed bed form, be about to polymeric adsorbent with about porous sieve plate be fixed in chromatographic column, make it and can't flow, be commonly called as " fixed bed ".

The characteristics of conventional fixed bed are that fluid is bigger by the resistance of bed because bed voidage is less, and fluid flows through bed and is " piston flow ", and the rate of flow of fluid that the footpath makes progress is uniform better.

But in the large-scale industrial production of the biochemical field of reality, because material liquid is generally the thick liquid that contains a large amount of solid particle/chips, the fixed bed chromatography column can can't directly be handled the material liquid that contains solid because easily get clogged, such as can't directly isolating target product from the high viscosity feed liquid that contains cell or cell fragment.Because the solid matter in the feed liquid can accumulate in the tiny runner, constantly increase bed resistance, finally stop up bed.

Therefore, in traditional handicraft/actual production, when adopting the fixed bed processing to contain solid material, need at first to adopt centrifuge to remove big slightly solid particle in the material, remove tiny particle with filter type more subsequently, enter the fixed bed chromatography column through pretreated material at last, carry out catching and separating of target composition.

Though traditional fixed bed chromatographic isolation technology can satisfy the separation requirement in the daily biogenetic products production process, but its technological process is longer, the equipment investment volume is higher, the capacity centrifuge costs an arm and a leg at a high speed/greatly, filter plant needs often to change filter membrane, caused the cost and the expense of whole production/operation higher, and be difficult to reduce.

Chromatography column adopts the notion of expanded bed (Expanded Bed Adsorption) to begin to release from the eighties in last century.

So-called expanded bed, be exactly that fluid enters bed from the chromatographic column bottom, the buoyancy and the surface drag that when utilizing fluid upwards to flow the solid particle adsorbent are produced, solid sorbent particles is upwards suspended in various degree and cause bed expansion, so be referred to as expanded bed, its concrete structure can be referring to shown in the Figure of description 1a.

Because solid sorbent particles is in suspended state in the expanded bed, the voidage of bed is much larger than the voidage of fixed bed, can make solid particle polluters such as cell in the material liquid, cell fragment smoothly by bed, be dissolved in target composition in the material liquid and be inflated in the bed solid absorbent and catch.Therefore, expanded bed need not the solid particle of material liquid is handled especially, can directly carry out chromatogram and catch; So, expanded bed can be applied in the chromatographic separation process, replacing traditional fixed bed chromatography column, and shortened process, reduce biochemical operating cost and the production cost that separates.

Though the expanded bed chromatography isolation technics is at the example of industrial existing successful Application, its scope of application is not extensive.At present, in the actual industrial large-scale production of biochemical field, have only a spot of commercial application, as Escherichia coli homogenate, inclusion body, Escherichia coli nutrient solution, yeast cells homogenate, Yeast Cultivation liquid, the extraction of Hybridoma Cell Culture liquid and animal tissue's product etc.

Influence one of chief reason of the expanded bed scope of application/service efficiency; be because expanded bed is when processing contains the feed liquid of solid; though the solid in the feed liquid can pass bed by the space between the adsorbent; but owing to stop/carry that the porous of adsorbent stops that the aperture of sieve plate/porous screen cloth gripper shoe (also claiming screen cloth gripper shoe or supporting screening plate) is less; solid particle can accumulate in the bottom that porous stops sieve plate or porous screen cloth gripper shoe; along with solid matter is poly-more many more, cause solid particle to stop up sieve plate and duct at last.And porous stops the obstruction in sieve plate and/or porous screen cloth gripper shoe and duct, gently then causes uniform fluid distribution to be worse off, and cleans difficulty, and is heavy then cause stopping production.

And in the material system that precipitation or reunion phenomenon are arranged, this type of stops up and not only takes place at the supporting screening plate position of expanded bed bottom, can stop that also the sieve plate position takes place in the porous at expanded bed top, so seriously influenced normally using and moving of expanded bed.

The subject matter that another influences the expanded bed scope of application/service efficiency is that traditional expanded bed distributor can not promptly form laminar flow in the bottom of expanded bed bed.

The fluid of expanded bed inside distributes can be referring to shown in the Figure of description 1b, and this figure has provided stream (body) the field distribution situation of expanded bed inside.As seen from the figure, because the bottom distributor of existing expanded bed can not form the equal compress water (being the even distribution of pressure and fluid) on the expanded bed cross section, cause the cloth water pressure of expanded bed central area and its fringe region unbalanced, and then caused non-flat current drainage form, make and the inner phenomenon of backflowing that produces of bed body caused the total number of theoretical plate lower (have only about 50) of expanded bed when absorption.

Number of theoretical plate is based on that flow pattern is meant in some lock out operation to finishing the quantity of the required theoretical tray of a certain appointment separation requirement near the parameter of laminar flow degree or axial backmixing degree in the measurement chromatographic column of time of staying notion.

Number of theoretical plate is the desirable laminar flow state of infinitely great representative, number of theoretical plate be 1 then correspondence the complete mixing flow state.

According to theory and empirical estimating, theoretical total plate number was greater than 400 o'clock, and the absorption/desorption performance of chromatographic separation device is subjected to the influence of plate number seldom again, and theoretical total plate number of in fact most of fixed bed far exceeds 400 (usually greater than 3000).The advantage of expanded bed is by sacrificing the ability that unnecessary number of theoretical plate improves processing flux and obtains direct processing particle, because the characteristic of its semi-fluidization, expanding bed itself can not provide very big resistance drop, so the liquid-distribution property of its distributor under low pressure drop has crucial influence to the expanded bed technology.

Present research about the expanded bed distributor seldom has open or appears in the newspapers and lead.

According to the knowledge of the applicant, had two to be mainly used in the technology that solves screen cloth and distributor obstruction: but the spider arm of the periodic rotary that Amersham Biosciences and Upfront propose is avoided blockage structure, and the sieve plate of mentioning among the U.S. Pat 2007199899A1 tangentially cleans the stream design.

Adopt the technical scheme (seeing Figure of description 2a) of spider arm configuration, its feeding liquid stream enters downwards in the expanded bed by the arm among the figure, and the spider arm that is made of a plurality of arms simultaneously periodically rotates, to try hard to alleviate the situation of its obstruction.

Though adopt spider arm design to solve the cleaning and the cleaning problems of chromatogram exchange column, but it uses the uniform poor-performing of liquid of back feed liquid, can not form laminar flow regularly in the bottom of expanded bed bed, can cause total number of theoretical plate of expanded bed on the low side (especially when desorption, causing the product of low resolution and dilution), lose the main advantage of chromatographic isolation.In addition, the rotation of spider arm has caused the interior frequent machinery of expanded bed exchange column to shake, and the mechanical seal difficulty has wearing and tearing to adsorption particle.Though, can in actual industrial production, not obtain large-scale promotion and application so this design has formed commercially produced product.

In U.S. Pat 2007199899A1, a kind of expanded bed splitter of energy automatically cleaning lower support sieve plate has been proposed, its concrete structure is seen shown in the Figure of description 2b, the sample feed liquid enters the bottom of expanded bed porous silk screen gripper shoe through feed pump 110 and flow controller 120, sample segment liquid passes porous silk screen gripper shoe and enters the polymeric adsorbent expanding bed, and more sample feed liquid, become vertical angle ground with expanded bed inner fluid direction, the lower surface of slipstream overexpansion bed porous silk screen gripper shoe (see among the figure shown in the arrow 125), thereby play the effect of cleaning, this strand sweeping fluid provides power and flow controller 130 control flows by circulating pump 140.

Adopt sieve plate tangentially to clean the stream design, can reduce the obstruction of porous silk screen gripper shoe in the distributor in theory and porous silk screen gripper shoe is played auxiliary cleaning action, but this design exist tangible deficiency:

1) can not consider the uniform pressure distribution problem that axially go up sample feed liquid of fluid, do not have to consider the uniform problem of (being on the sieve plate plane) fluid pressure on the direction perpendicular yet with importing and exporting axis from import to outlet.Therefore, the feed liquid fluid in this distributor all can have in various degree pressure inhomogeneous (depending on the flow velocity of liquid purge and the volume of distributor cavity) on the axis of outlet and the perpendicular axis thereof in import.In order to reach the target of uniform pressure distribution, the cavity volume of expanded bed certainly will be wanted greatly, but the volume of cavity increased, and can cause air-teturning mixed phenomenon serious again, influences the result of use/service efficiency of expanded bed;

2) arrived " going up sample " process in, late period, along with the increase of solid content in the circulation sample liquid, solid particle generation coalescence, the circulation sample liquid of high viscosity, high solids content is difficult to play the effect of cleaning porous silk screen gripper shoe at this moment, clogging still can take place, and is difficult to avoid.

So this technical scheme does not obtain practical application up at present in real suitability for industrialized production, also not seeing has corresponding commercial product.

Summary of the invention

Technical problem to be solved by this invention provides a kind of expanded bed chromatography splitter and technological process that is used for biochemical separation processes, its by stop in expanded bed bottom porous sieve plate below the liquid with self-cleaning function be set all press distributor, the upper nozzle assembly that has the recoil cleaning function in the setting of expanded bed capital end, make the expanded bed chromatography splitter be applicable to and handle the raw material feed liquid that contains solid particle polluter, simultaneously, can carry out uniform to the material sample feed liquid that enters expanded bed better, solved expanded bed in the industry's enlarging production process under the low bed layer pressure drop feed liquid liquid supply pressure along the equally distributed problem of radial direction, guaranteed preferably to present the laminar flow state by the fluid of expanded bed bed, improve the result of use and the service efficiency of expanded bed greatly, improve the absorption carrying capacity and the desorption resolution ratio of expanded bed, also can realize " dress post on the throne " and " post that unloads on the throne " operating procedure of expanded bed splitter easily.

Technical scheme of the present invention is: a kind of expanded bed chromatography splitter that is used for biochemical separation processes is provided, comprise that the porous that is positioned at expanded bed capital end stops sieve plate and discharge nozzle, be positioned at the porous supporting screening plate and the feed pipe of expansion column bottom, it is characterized in that: on the top of expanded bed, be provided with and comprise and go out pipe, solid dirt delivery pipe, mobile stick harness and sealing ring on the feed liquid the having upper nozzle assembly of the cleaning function that recoils at least; In the bottom of described expanded bed, be provided with and comprise feed pipe, radial spray hole, comb under water conservancy diversion disk and the feed liquid radially at least, the liquid with self-cleaning function is all pressed distributor.

When porous stops the sieve plate below solid particle accumulation is arranged, cause porous to stop when the resistance drop of sieve plate increases, backwash liquid goes out pipe from feed liquid and enters expanded bed capital end, porous is stopped the sieve plate cleaning that recoils, blow off accumulating in the solid accumulation that porous stops the sieve plate bottom, move down mobile stick harness simultaneously, open the solid dirt delivery pipe, the solid accumulation is discharged from the solid dirt delivery pipe; After recoil finishes, on move mobile plunger, close the solid dirt delivery pipe, expanded bed is proceeded the operation of normal chromatographic isolation.

Described radially water conservancy diversion disk cooperates with the lower surface of porous screen cloth gripper shoe, on the radial direction of water conservancy diversion disk, constitutes variable cross-section uniform fluid distribution runner at the center of water conservancy diversion disk radially; The raw material feed liquid that from described radial spray hole, ejects, along the expanded bed or the radial direction of water conservancy diversion disk radially, to spraying all around, under the guiding of water conservancy diversion disk radially, the porous supporting screening plate is carried out side direction/tangentially wash away, the solid particle flushing reflux main flow that will assemble in the sieve plate bottom, comb is discharged under the feed liquid, turn back in the sample circulating tank and circulate, realize cleaning complete on the throne and automatic cleaning action the porous supporting screening plate; The upper nozzle assembly and the liquid that have recoil cleaning and/or self-cleaning function by setting are all pressed distributor, make the expanded bed chromatography splitter be applicable to and handle the raw material feed liquid that contains solid particle polluter, improve/improve the result of use/service efficiency of expanded bed in the industrial-scale production of biochemical field with this.

Concrete, on the one hand, go out pipe and solid dirt delivery pipe on the described feed liquid and be arranged on the expanded bed wall that porous stops the sieve plate top, described solid dirt delivery pipe runs through the middle part setting that expanded bed upper end wall and porous stop sieve plate; Described mobile stick harness and sealing ring are arranged in the solid dirt delivery pipe.

When moving on the described mobile stick harness, it cooperates with sealing ring, closes the solid dirt delivery pipe, and described upper nozzle assembly is in normal operating conditions, and the raw material feed liquid in the expanded bed is flowed through after porous stops sieve plate, discharges by going out pipe on the feed liquid; When described mobile stick harness moves down, the conducting of solid dirt delivery pipe, described upper nozzle assembly are in recoil cleaning state, and backwash liquid is through going out to manage reverse flow after porous stops sieve plate on the feed liquid, to accumulate in porous and stop that sieve plate bottom solid accumulation blows off, and is discharged by the solid dirt delivery pipe; After the recoil cleaning finishes, move on the mobile stick harness, close the solid dirt delivery pipe, can proceed normal chromatographic isolation operation; Simultaneously, utilize mobile stick harness in the upper nozzle assembly on move or move down, close or open the solid dirt delivery pipe, be used for realizing the filling on the throne and/or the unloading of solid absorption particle in the expanded bed.

Concrete, on the other hand, feed pipe being set in the centre of described expansion column bottom, the top of feed pipe is arranged on the below of porous supporting screening plate, on the top of described feed pipe the radial spray hole is set; Between expansion column bottom wall and porous supporting screening plate, radially water conservancy diversion disk is set; Offer through hole in the centre of water conservancy diversion disk radially, described feed pipe runs through the radially through hole setting of water conservancy diversion disk; Described radially water conservancy diversion disk cooperates with the lower surface of porous supporting screening plate, on the radial direction of water conservancy diversion disk, constitutes variable cross-section uniform fluid distribution runner at the center of water conservancy diversion disk radially; Comb is arranged on the expanded bed wall of porous supporting screening plate below under the described feed liquid.

Described variable cross-section uniform fluid distribution runner is radially variable cross-section fluid course or radially variable mass circulation road, it is to entering the raw material feed liquid of expanded bed, carry out along expanded bed fluid pressure radially uniform, guarantee the fluid pressure substantially constant radially under the porous screen cloth gripper shoe, thereby make feed liquid fluid present the laminar flow state by the expanded bed bed, realize cleaning on the throne and automatically cleaning simultaneously, improve/improve the result of use/service efficiency of expanded bed with this.

Further, at the upper surface of described radially water conservancy diversion disk, be provided with the deflector that polylith is erect along radial direction, described deflector is pressed the uniform arrangement setting of circumferencial direction; Below the described radially water conservancy diversion disk and between the wall of expanded bed lower end, the runner that backflows is set; The diameter of described radially water conservancy diversion disc centre position through hole between through hole and feed pipe outer tube wall, forms a liquid communication gap more than or equal to the external diameter of described feed pipe, constitutes the internal liquid circulatory flow.

The present invention also provides a kind of expanded bed chromatography splitter that is used for biochemical separation processes, comprise the expansion column, top at described expansion column, be provided with and go out pipe and solid dirt delivery pipe on the feed liquid, in the bottom of described expansion column, comb under feed pipe and the feed liquid is set, described porous stops that going out pipe, solid dirt delivery pipe and mobile stick harness on sieve plate, expanded bed upper end wall, the feed liquid constitutes the column cap assembly, be arranged on the upper end of expansion column movably, it is characterized in that:

Sample liquid circulating tank, buffer solution circulating tank are set, unload post solution circulating tank, the adsorbent tank that has the centrifugal solid-liquid separator, feed pump, dress post pump and online coarse filter, constitute a chromatographic isolation unit that is applicable to biochemical extractor gauge modelling production process with described expansion column.

Wherein, the outlet of described sample circulating tank and buffer solution circulating tank is connected with the feed pipe of expanded bed through feed pump; Go out pipe on the feed liquid of described expanded bed and be connected with the product jar, perhaps, go out pipe on the feed liquid and be connected with waste water or intermediate storage jar through capital outlet pressure control valve through capital outlet pressure control valve.

Comb is connected with the liquid back pipe of online coarse filter with the sample circulating tank through the circular flow pressure-regulating valve under the feed liquid of described expanded bed, constitutes closed circuit; Comb is connected through the liquid back pipe of circular flow pressure-regulating valve with the buffer solution circulating tank under the feed liquid of described expanded bed.

Going out on the feed liquid of described expanded bed under pipe and the feed liquid between the comb, bypass conduit is set.

Going out pipe on the feed liquid of described expanded bed capital end is connected with the outlet of feed pump through valve.

The solid dirt delivery pipe of described expanded bed passes through upper inlet pipe, the upper outlet pipe of adsorbent tank and unloads post solution circulating tank, is connected with the inlet tube of feed pump.

The following outlet of described adsorbent tank is connected with the solid dirt delivery pipe of expanded bed through dress post pump.

First atmospheric valve is set on comb road under the feed liquid of described expanded bed; Second atmospheric valve is set on the feeding pipe of described expanded bed; On the solid dirt discharge pipe of described expanded bed, the 3rd atmospheric valve is set.

On the pipe of the import/export of described expanded bed or each jar, valve is set; Inlet flow rate meter, inlet pressure meter and online conductivity meter are set on the feed pipe of expanded bed, on the feed liquid of expanded bed, go out to be provided with on the pipe online conductivity meter, pH meter, rate of discharge meter and ultraviolet tester, on the connecting line between expanded bed and the sample circulating tank/buffer solution circulating tank return duct, the circulating fluid pressure meter is set.

When described expanded bed was in normal operating conditions, the raw material feed liquid in the sample circulating tank was sent into through feed pump and is carried out chromatographic isolation in the expanded bed, and the feed liquid after the separation goes out pipe output on feed liquid; The raw material feed liquid of comb circulating reflux is returned the sample circulating tank again after online coarse filter filters under the feed liquid.

When described expanded bed was in recoil cleaning state, backwash liquid was pumped into the top of expanded bed through the dress post, and the porous that is positioned at the expanded bed upper end is stopped the sieve plate cleaning that recoils; Simultaneously, the mobile stick harness in the solid dirt delivery pipe moves down, and opens top jet nozzle outlet, takes the solid particle that gathers under the porous screen cloth out of expanded bed and discharges from the solid dirt delivery pipe.

After the recoil cleaning finishes, move on the mobile stick harness in the solid dirt delivery pipe, close the solid dirt delivery pipe, proceed normal chromatographic isolation operation.

Simultaneously, utilize mobile stick harness in the upper nozzle assembly on move or move down, close or open the solid dirt delivery pipe, under the cooperation of unloading post solution circulating tank, the adsorbent tank that has whizzer, feed pump and dress post pump, realize the filling on the throne and/or the unloading of solid absorption particle in the expanded bed.

Further, described sample circulating tank is connected with sample jar, cleaning fluid tank and/or buffering flow container, and the liquid in described sample jar, cleaning fluid pipe and/or the buffering flow container adopts the mode of liquid level control to mend in the sample circulating tank.

Described buffer solution circulating tank is connected with the buffering flow container, and the liquid in the described buffering flow container adopts the mode of liquid level control to mend in the buffer solution circulating tank.

Described charging pump intake piping is connected with the outlet of wash-out flow container.

At the outlet of described feed pump, be provided for catching and removing the air trap of bubble.

The present invention also provides a kind of above-mentioned chromatographic isolation technology that is used for the expanded bed chromatography splitter of biochemical separation processes, it is characterized in that the chromatographic isolation technological process of described expanded bed chromatography splitter comprises the following steps: at least

A, chromatographic column balance:

The feed pipe of expanded bed bottom squeezed into buffer solution by feed pump from the buffer solution circulating tank, buffer solution goes out the pipe and the first atmospheric valve emptying earlier on feed liquid, switch to the buffer solution circulating tank again, to form suitable circular flow and to cross the post flow;

Described post flow and the circular flow crossed, the feedback of the inlet flow rate meter bottom expanded bed and the rate of discharge meter at top, nationality is realized with pump speed, circular flow pressure-regulating valve and the capital outlet pressure control valve of control feed pump;

After flow pressure is stable, allow chromatographic column pass through a certain amount of buffer solution, make ultraviolet tester, pH meter or online conductivity meter instrumentation be stabilized to the technology specified scope, promptly finish the chromatographic column balance according to technological requirement.

Sample on B, the chromatographic column;

The feed pump import switches to sample liquid circulating tank outlet, and comb switches to sample liquid circulating tank inlet tube under the feed liquid of loop exit expanded bed bottom; Continuation is by inlet flow rate meter and rate of discharge meter feedback and regulate feed pump, circular flow pressure-regulating valve and capital outlet pressure control valve, controls required post flow and circular flow excessively;

The sample feed liquid is squeezed into the expanded bed inlet tube through feed pump in the sample circulating tank, wherein a part of feed liquid by comb circulating reflux under the feed liquid of expanded bed bottom to the sample circulating tank, another part feed liquid is passed the porous supporting screening plate and is entered expanding bed, by the absorbent particles in the expanded bed target component in the feed liquid is adsorbed;

The porous that the feed liquid that has been adsorbed the target component is passed expanded bed top stops that sieve plate flows out expanded bed; Discharge system or enter the intermediate storage jar after ultraviolet tester, pH meter or online conductivity meter detect again;

Fresh sample liquid is constantly mended to the sample circulating tank in the mode of liquid level control from sample jar;

During the last sample,, can suspend and go up the sample operation if obvious rising falls in expanded bed chromatography splitter pressure, the feed pump import is switched to the buffer solution circulating tank, close the feed pipe of expanded bed bottom, open bypass, buffer solution is introduced the expanded bed top, and the porous that the top is cleaned in recoil stops sieve plate; Mobile simultaneously stick harness moves down, and opens top upper nozzle assembly, porous is stopped the solid material that gathers under the sieve plate is taken expanded bed out of by the solid dirt delivery pipe and from the emptying of waste liquid mouth;

Recoil is closed feed pump after cleaning and finishing, and moves on the mobile stick harness, closes top upper nozzle assembly, closes the top bypass, opens expanded bed bottom sample introduction pipe, comes back to the above-mentioned sample operation of going up again;

Intact etc. sample treatment, last sample promptly comes to an end.

C, chromatographic column flushing;

Behind the end of the sample, the feed pump import switches to the buffer solution circulating tank, and the expanded bed circulation port switches to waste liquid emptying earlier, switches back to the buffer solution circulating tank again; Control same post flow and circular flow excessively, allow chromatographic column pass through a certain amount of buffer solution, make ultraviolet tester, pH meter or online conductivity meter instrumentation be stabilized to the technology specified scope and promptly finish the chromatographic column flushing according to technological requirement.

D, adsorbate wash-out;

At first stop feed pump, allow the expanded bed bed leave standstill, described column cap assembly is reduced to bed leave standstill interface and fixing, make chromatographic column become the fixed-bed structure state by the expanded bed configuration state;

Then, restart feed pump, the inlet tube of feed pump is cut to the buffer solution circulating tank, and expanded bed chromatography splitter circulation port is closed, and crosses the post flow by bottom inlet flowmeter FEEDBACK CONTROL; Allow the expanded bed chromatography splitter by a certain amount of buffer solution according to technological requirement, make ultraviolet tester, pH meter or online conductivity meter instrumentation be stabilized to the technology specified scope;

During wash-out, the feed pump import switches to the outlet of wash-out flow container and is connected, and expanded bed chromatography splitter loop exit is still closed; Continuation is crossed the post flow by bottom inlet flowmeter FEEDBACK CONTROL;

Monitoring ultraviolet tester, pH meter and/or online conductivity meter, the machine switching is introduced the product jar with target peak when appropriate, and all the other put into waste liquid or middle remaining tank.

E, chromatographic column are cleaned and regeneration;

After wash-out is finished, stop feed pump, the top porous is stopped the upper nozzle assembly of sieve plate is opened, and by the logical atmosphere of drain;

Then described column cap assembly is risen to original height, fixedly the column cap assembly makes chromatographic column revert to the expanded bed configuration state, moves on the mobile stick harness, closes the upper nozzle assembly;

With the sample whole emptying of pot liquid that circulate, cleaning fluid self-cleaning flow container is introduced the sample circulating tank; Start feed pump, the feed pump import is switched to the sample circulating tank, the chromatographic column circulation port switches counter sample product circulating tank;

According to technological requirement, control required post flow and the circular flow crossed.Allow chromatography column pass through a certain amount of cleaning fluid, make ultraviolet tester, pH meter or online conductivity meter instrumentation be stabilized to the technology specified scope, promptly finish the cleaning and the regeneration of chromatographic column.

F, the heavy balance of chromatographic column;

After chromatographic column is cleaned and regeneration finishes, close feed pump,, close each emptying valve, buffer solution is introduced the sample circulating tank from cushioning flow container the sample whole emptying of pot liquid that circulate;

Start feed pump, the feed pump import is switched to the sample circulating tank;

The chromatographic column circulation port is earlier from the emptying of waste liquid mouth, switchback sample circulating tank again;

According to technological requirement, with required post flow and the circular flow crossed of above-mentioned same method control;

Allow chromatographic column pass through a certain amount of buffer solution, make ultraviolet tester, pH meter or online conductivity meter instrumentation be stabilized to the technology specified scope, promptly finish the heavy balance of chromatographic column;

Close down the feed pump system, each emptying valve is closed in the whole emptying of sample circulation pot liquid, waits for the next operation circulation.

Further, the above-mentioned chromatographic isolation technology that is used for the expanded bed chromatography splitter of biochemical separation processes, its described chromatographic isolation technological process also comprises the following steps:

G, dress post:

At first, earlier the expanded bed chromatography splitter is filled with buffer solution, again the upper nozzle assembly outlet at expanded bed top is opened;

Start dress post pump, absorbent particles suspension is squeezed into the chromatographic column from adsorbent tank;

The adsorption particle natural subsidence is in chromatographic column, and liquid stops that through porous sieve plate brings out a mouthful outflow from the expanded bed capital, realizes dress post on the throne.

H, unload post:

At first, go out pipe on the feed liquid at expanded bed top and close, the top is moved stick harness and is moved down;

The feed pump import switches to unloads post buffer solution circulating tank;

Start feed pump and suitably open big flow, make the fluid velocity in the expanded bed surpass the adsorption particle sinking speed, absorbent particles can be brought into adsorbent tank;

Wherein, the centrifugal solid-liquid separator in the adsorbent tank is stayed the adsorption particle medium in the jar, and solution then overflows and recycles;

Continue the operation feed pump and all be moved in the adsorbent tank, realize the post that unloads on the throne until all adsorption particle media.

More specifically, in described " going up sample " the step later stage, can to reduce the viscosity and the granule density of raw material feed liquid, improve the rate of recovery by in the sample circulating tank, injecting dilution.

More specifically, described circular flow and the ratio of crossing the post flow depend on the needs that prevent to block with uniform pressure distribution between 0 to 10.

Compared with the prior art, advantage of the present invention is:

By stop in expanded bed bottom porous sieve plate below the liquid with self-cleaning function be set all press distributor, have the upper nozzle assembly of recoil cleaning function in the setting of expanded bed capital end, make the expanded bed chromatography splitter be applicable to and handle the raw material feed liquid contain solid particle polluter;

2. the water conservancy diversion disk constitutes the feed liquid distributor with the radial spray hole that is positioned at the feed pipe end by being provided with radially, its radial flow and specific uniform fluid distribution runner/distribution channel design, guaranteed that the feed liquid fluid can have higher/even flow from the center to the edge, make the feed liquid fluid pressure under the sieve plate be evenly distributed, and it is high more that feed liquid is sprayed flow velocity, it is even more to distribute, and has brought the uniform effect of feed liquid fluid pressure of self-stabilization;

3. radially on the water conservancy diversion disc radius direction, the radially longitudinal section of fluid passage is designed to the wedge shape version of " center is thick; thin edge ", constitute radially a variable cross-section fluid course or a variable mass circulation road radially, circulation area when radially flowing by changing fluid, overcome because of variable mass and flow and runner is fan-shaped geometry and changes caused pressure and change, to guarantee the fluid pressure substantially constant radially under the porous sieve plate, thereby the fluid by bed presents the laminar flow state, so can improve the separation theorem plate number and the chromatographic isolation efficient of expanded bed greatly;

4. distributor is provided with comb under fluid feed pipe and the feed liquid except that porous screen cloth/sieve plate, can be used with external pump, feed liquid is circulated between storage tank and distributor, and this circular flow directly influences the feed liquid of feed pipe and sprays flow velocity, can be used as the performance variable that controlled pressure distributes;

Since fluid course size and porous screen cloth/sieve plate resistance when design, determined, circular flow and to see through flow then be the controllable operating variable, so the whole control easily of its expanded bed, the industrially scalable amplification is easy; Simultaneously, dead volume is little in the distributor, the mixed bed of distributor internal return is compared and can be ignored, thereby can realize the uniform fluid distribution in the small size under the low pressure, make the industrial applications of expanded bed on biochemical isolation technics of low thermal expansion, high number of theoretical plate be achieved, reach the core advantage of using expanded bed;

6. be provided with the internal liquid circulatory flow, spray flow velocity when enough high from feed pipe, can form inner partial circulating in the distributor, further guaranteed the pressure substantially constant radially under porous screen cloth/sieve plate when feed liquid.Thereby the fluid by bed presents the laminar flow state, so, can improve the absorption carrying capacity and the desorption resolution ratio of expanded bed further;

7. all press distributor and upper nozzle assembly by liquid with recoil cleaning function with self-cleaning function, in conjunction with external equipment, realize " on the throne " dress post of expanded bed and unloaded post, improved operating efficiency greatly, can effectively improve/improve the result of use/service efficiency of expanded bed.

Description of drawings

Fig. 1 a is the structural representation of existing expanded bed;

Fig. 1 b is the fluid distribution schematic diagram of existing expanded bed inside;

Fig. 2 a is the schematic diagram of spider arm technical scheme;

Fig. 2 b is the schematic diagram that the United States Patent (USP) sieve plate tangentially cleans the stream design;

Component locations and flow direction schematic diagram when Fig. 3 a is a upper nozzle assembly operate as normal of the present invention;

Component locations and flow direction schematic diagram when Fig. 3 b is upper nozzle assembly recoil cleaning;

Fig. 4 a is structure and the flow direction schematic diagram that liquid of the present invention is all pressed distributor;

Fig. 4 b is the radially plan structure and the flow direction schematic diagram of water conservancy diversion disk of the present invention;

Fig. 5 is the internal flow distribution schematic diagram of expanded bed in the technical program;

Fig. 6 is the equipment connection structure schematic diagram of chromatographic isolation unit in the technical program;

Fig. 7 is the schematic diagram of technological process in the technical program;

Fig. 8 is the technological operation step schematic diagram of expanded bed;

Fig. 9 is a dress post and the flow direction schematic diagram that unloads post in technological process.

102 is the feed liquid storage tank among the figure, and 110 is feed pump, and 120 is flow control component, and 125 is slipstream, and 130 is flow control component, and 140 is the circular flow pump, and 202 is splitter, and 220 is the expanded bed adsorption resin, and 222 is the lower support sieve plate.

301 for porous stops sieve plate, and 302 is expanded bed upper end wall, and 303 for going out pipe on the feed liquid, and 304 is the solid dirt delivery pipe, and 305 is mobile stick harness, and 306 is sealing ring, and 307 is the raw material feed liquid, and 308 is backwash liquid.

401 is the expanded bed wall, and 402 is porous screen cloth gripper shoe, and 403 is feed pipe, 404 is the radial spray hole, 405 are water conservancy diversion disk radially, and 405-1 is the centre of water conservancy diversion disk radially, and 405-2 is the circumferential edges position of radially water conservancy diversion disk, 405-3 is a through hole, 405-4 is to backflow in water conservancy diversion disk below, and 406 is comb under the feed liquid, and 407 are variable cross-section tangent line stream, 408 is the internal liquid circular flow, and 409 is deflector.

V101 is the sample liquid circulating tank, V102 is the buffer solution circulating tank, V103 is for unloading post solution circulating tank, V104 is an adsorbent tank, and LXFLQ is the centrifugal solid-liquid separator, and T101 is expanded bed chromatography splitter (abbreviating expanded bed or exchange column as), B101 is a feed pump, B102 is dress post pump, and F101 is the circular flow pressure-regulating valve, and F102 is a capital outlet pressure control valve, ZXCLQ is online coarse filter, PIT-1 is the inlet pressure meter, and PIT-2 is the circulating fluid pressure meter, and FIT-1 is the rate of discharge meter, FIT-2 is the inlet flow rate meter, KQXJ is the air trap, and YPG is a sample jar, and QXYG is a cleaning fluid tank, XTYG is the wash-out flow container, HCYG is the buffering flow container, and CPG is the product jar, and Cond is online conductivity meter, PH is a pH meter, UV is the ultraviolet tester, F051, F052 and F014 are followed successively by first, the second and the 3rd atmospheric valve, all the other F001～F017 are valve.

The specific embodiment

The present invention will be further described below in conjunction with drawings and Examples.

Among Fig. 1 a, fluid enters bed from the bottom of chromatographic column, and the buoyancy and the surface drag that when utilizing fluid upwards to flow the solid particle adsorbent are produced upwards suspend solid sorbent particles in various degree and cause bed expansion, so be referred to as expanded bed.

Because solid sorbent particles is in suspended state in the expanded bed, the voidage of bed is much larger than the voidage of fixed bed, can make solid particle polluters such as cell in the material liquid, cell fragment smoothly by bed, be dissolved in target composition in the material liquid and be inflated in the bed solid absorbent and catch.

Owing to stop/carry that the porous of adsorbent stops that the aperture of sieve plate/porous screen cloth gripper shoe (also claiming screen cloth gripper shoe or supporting screening plate) is less in the expanded bed; solid particle can accumulate in the bottom that porous stops sieve plate or porous screen cloth gripper shoe; along with solid matter is poly-more many more, cause solid particle to stop up sieve plate and duct at last.

In addition, traditional expanded bed distributor can not promptly form laminar flow in the bottom of expanded bed bed, has caused the total number of theoretical plate of expanded bed when absorption lower.

Among Fig. 1 b, because the bottom distributor of conventional expanded bed can not form the equal compress water on the expanded bed cross section, caused the cloth water pressure of expanded bed central area and its fringe region unbalanced, and then caused non-flat current drainage form, make and the inner phenomenon of backflowing (flowing expression with annulus among the figure) that produces of bed body caused the total number of theoretical plate of expanded bed when absorption lower.

Among Fig. 2 a, feeding liquid stream enters in the expanded bed downwards by several arms among the figure, the spider arm rotation regularly/regularly that is made of several arms simultaneously, the lower support silk screen/sieve plate that expanded bed is carried adsorbent washes accordingly/washes away, to try hard to alleviate the situation of its obstruction.

Though adopt spider arm design to solve the cleaning and the cleaning problems of pillar, it can cause total number of theoretical plate of expanded bed on the low side (especially causing the product of low resolution and dilution when desorption) after using, and has lost the main advantage of chromatographic isolation.

In addition, the rotation of spider arm has caused the interior frequent machinery of expanded bed exchange column to shake, and the mechanical seal difficulty has wearing and tearing to adsorption particle.

Among Fig. 2 b, the sample feed liquid enters the below of the porous silk screen gripper shoe 222 of expanded bed splitter 202 bottoms through feed pump 110 and flow control component 120, sample segment liquid passes the expanding bed that porous silk screen gripper shoe enters polymeric adsorbent 220, and more sample feed liquid, become vertical angle ground with expanded bed inner fluid direction, the lower surface (seeing slipstream 125 indications among the figure) of slipstream overexpansion bed porous silk screen gripper shoe, thereby play the effect of cleaning, the tangential sweeping fluid of this strand is provided power and is controlled flow by flow controller 130 by circulating pump 140.

The feed liquid fluid of this design all can have in various degree liquid supply pressure non-uniform phenomenon (size that depends on the flow velocity and the distributor cavity volume of liquid purge) on the axis of outlet and the perpendicular axis thereof in import; In order to reach the uniform target of liquid supply pressure, the cavity volume of expanded bed certainly will be wanted greatly, but the volume of cavity increased, and can cause air-teturning mixed phenomenon serious again, influences the result of use/service efficiency of expanded bed.

Simultaneously, along with the increase of solid content in the circulation sample liquid, solid particle generation coalescence, the circulation sample liquid of high viscosity, high solids content is difficult to play the effect of cleaning porous silk screen gripper shoe at this moment, and clogging still can take place, and is difficult to avoid.

Among Fig. 3 a, the application's technical scheme provides a kind of expanded bed chromatography splitter that is used for biochemical separation processes, and one of its inventive point is:

On the top of expansion column 302, be provided with and comprise and go out pipe 303, solid dirt delivery pipe 304, mobile stick harness 305 and sealing ring 306 on the feed liquid the having upper nozzle assembly of recoil cleaning function at least.

Wherein, going out pipe and solid dirt delivery pipe on the feed liquid is arranged on the expanded bed wall 302 that porous stops sieve plate 301 tops, the solid dirt delivery pipe runs through expanded bed upper end wall and porous stops that the middle part of sieve plate is provided with, and mobile stick harness and sealing ring are arranged in the solid dirt delivery pipe.

When moving to as shown in this figure the position on the mobile stick harness, mobile stick harness cooperates with sealing ring, closes the solid dirt delivery pipe, and the upper nozzle assembly is in " operate as normal " state, raw material feed liquid 307 in the expanded bed is flowed through after porous stops sieve plate, discharges by going out pipe on the feed liquid.

Among Fig. 3 b, when porous stops the sieve plate below solid particle accumulation is arranged, cause porous to stop when the resistance drop of sieve plate increases, mobile stick harness is displaced downwardly to position shown in this figure, solid dirt delivery pipe conducting/be opened, and the upper nozzle assembly is in " recoil is cleaned " state, backwash liquid 308 goes out pipe from feed liquid and enters expanded bed capital end, reverse flow will accumulate in porous and stop that sieve plate bottom solid accumulation blows off, and is discharged by the solid dirt delivery pipe after porous stops sieve plate.

After the recoil cleaning finishes, move on the mobile stick harness, close the solid dirt delivery pipe, can proceed normal chromatographic isolation operation.

Among Fig. 4 a, another inventive point of the application's technical scheme is: below the porous screen cloth gripper shoe of expansion column lower end, be provided with one and comprise that end has the feed pipe in radial spray hole and the radially water conservancy diversion disk uniform effect of feed liquid that constituted, that have self-stabilization and have the even distributor of feed liquid of self-cleaning function.

On frame for movement, its feed pipe 403 is arranged on the centre of expansion column bottom 401 walls, and the end of feed pipe is arranged on the below of porous screen cloth gripper shoe 402, at the end of feed pipe, is provided with radial spray hole 404; Between expansion column bottom wall and porous screen cloth gripper shoe, a water conservancy diversion disk 405 radially is set; Offer through hole 405-3 in the centre of water conservancy diversion disk radially, the diameter of through hole is more than or equal to the external diameter of described feed pipe, and feed pipe runs through the through hole setting.

Radially the water conservancy diversion disk cooperates with the lower surface of porous screen cloth gripper shoe, at the center of water conservancy diversion disk radially on the radial direction of water conservancy diversion disk, constituted a uniform fluid distribution runner and (seen that the variable cross-section tangent line flows path, 407 place among the figure, for succinctly also available 407 representing this uniform fluid distribution runner, down with); Its uniform fluid distribution runner along the water conservancy diversion disk diameter to the longitudinal cross-section be wedge shape structure.

Below water conservancy diversion disk radially and between the wall of expanded bed lower end, be provided with the runner that backflows (see among the figure path, 405-4 place of backflowing, water conservancy diversion disk below, for succinct also available 405-4 represents this runner that backflows, down with).

The diameter of its through hole is greater than the external diameter of feed pipe, between through hole and feed pipe outer tube wall, form a liquid communication gap, constitute the internal liquid circulatory flow and (see path, internal liquid circular flow 408 place among the figure, for succinctly also available 408 representing this internal liquid circulatory flow, down with).

On the wall of expansion column bottom, be provided with comb 406 under the feed liquid.

Wherein, feed pipe is along the longitudinal axis setting of expanded bed; The radial spray hole is provided with along the tube wall circumference uniform distribution of feed pipe end.

Concrete, radially the water conservancy diversion disk is for being inverted horn-like or being inverted the taper disk, distance between the centre 405-1 of described radially water conservancy diversion disk and porous screen cloth gripper shoe 402 lower surfaces is greater than radially the circumferential edges position 405-2 of water conservancy diversion disk and the distance between porous screen cloth gripper shoe 402 lower surfaces.

Because there is pressure reduction in porous supporting screening plate 402 both sides, part material liquid and tiny solid particle thereof can pass the porous supporting screening plate and enter in the adsorption bed of expansion, and then have realized the absorption of target composition.

From prior art as can be known, when material liquid after jet apertures ejection, in water conservancy diversion disk circumference edge flow process, have part material liquid can pass the porous supporting screening plate and enter in the expanded bed, so the volume flow of fluid and mass flow are all changing.Simultaneously, liquid by process of flowing around the middle mind-set in because runner is fan-shaped, circulation area progressively enlarges, can produce fluid along pressure distribution on the radial direction or pressure differential, make the pressure reduction of porous supporting screening plate both sides have a undesirable distribution along radial direction.

On the one hand, for avoiding this unfavorable pressure distribution to change, uniform fluid distribution runner longitudinal cross-section radially is designed to the version that is the wedge shape of " in not lend oneself to worry and anxiety/thick, thin edge/narrow " in the technical program, circulation area when radially flowing by changing fluid overcomes because of variable mass and flows and runner is fan-shaped geometry and changes caused cloth liquid/liquid supply pressure and change.

In addition, above-mentioned uniform fluid distribution runner adopts the longitudinal cross-section version of wedge shape, make that radially the water conservancy diversion disk matches with the lower surface of porous screen cloth gripper shoe, below porous screen cloth gripper shoe, constitute radially a variable cross-section fluid course or a variable mass circulation road radially, on the path flowed through from the center of water conservancy diversion disk along the outside liquid of its radial direction, the cross-sectional area of flow channel for liquids/passage successively decreased successively, and the circulation area of fluid when the water conservancy diversion disk is made Radial Flow reduces successively.

Like this, the technical program is by said structure and layout designs, circulation area when changing fluid along water conservancy diversion disk Radial Flow, overcome/offset because of variable mass and flow and runner is fan-shaped geometry and changes caused pressure and change, guaranteed that the feed liquid fluid can have higher/even flow from the center to the edge, made that the feed liquid fluid pressure under the sieve plate is evenly distributed, and feed liquid injection flow velocity is high more, it is even more to distribute, and has brought the uniform effect of feed liquid liquid of self-stabilization.

By the setting runner that backflows, the pressure of having guaranteed liquid under the porous sieve plate is substantially constant radially.

On the other hand, when the material sample feed liquid from the radial spray hole of feed pipe end, along the expanded bed or the radial direction of water conservancy diversion disk radially, to around when spraying, under the guiding of water conservancy diversion disk radially, porous screen cloth gripper shoe is carried out side direction/tangentially wash away, realization is to the automatic cleaning action of porous screen cloth gripper shoe, avoid the sieve plate obstruction, can realize complete cleaning on the throne, no dead point/dead band.

At last, the material sample feed liquid is turned back through the runner that backflows downwards at the circumferential edges position of water conservancy diversion disk radially, the solid particle flushing reflux main flow that will below porous screen cloth gripper shoe, assemble, and comb is discharged under the feed liquid, turns back to and carries out outer circulation in the head tank.

Under the actual working state, the material sample feed liquid enters distributor by feed pipe 403, from the spray-hole 404 at the terminal top of feed pipe along radial direction, to spraying all around, uniform fluid distribution runner 407, radially the water conservancy diversion disk circumference edge 405-2 and the runner 405-4 that backflows successively flow through, comb 406 is discharged under the feed liquid at last, turns back to and carries out outer circulation in the recycle feed flow container.

As further extension, in the closed circuit coarse filter or screen cloth can also be set outside, to remove the agglomerate solids that forms in the processing procedure, can in material liquid tank, inject dilution, with the viscosity and the raising rate of recovery that reduces feed liquid in " going up sample " later stage.

Among Fig. 4 b, radially water conservancy diversion disk 401 is being provided with the deflector 409 that polylith is erect along radial direction, and deflector press the uniform arrangement setting of circumferencial direction,, can evenly distribute along circumference when the circumferential edges direction is mobile with the fluid guaranteeing to eject from spray-hole.

Among Fig. 5, by adopting the structure and layout design shown in earlier figures 4a and Fig. 4 b, radially the water conservancy diversion disk structurally matches with porous screen cloth gripper shoe, utilize the radially form of uniform pressure distribution of the variation of fluid course radial section or fluid, the circulation canal that is aided with internal liquid, carry out along expanded bed liquid radially uniform to the material sample feed liquid that enters expanded bed, guaranteed the fluid pressure substantially constant radially under the porous screen cloth gripper shoe, thereby make fluid present the laminar flow state by the expanded bed bed, improve the separation theorem plate number and the chromatographic isolation efficient of expanded bed, the result of use/service efficiency of improvement/raising expanded bed with this.

The parallel liquid stray arrow head that makes progress in the expanded bed among the figure, just shown the laminar flow that is independent of bed resistance in the chromatogram exchange column, it is compared with the non-ideal flow form shown in Fig. 1 b, can obviously find out, because the cloth water pressure substantially constant of expanded bed central area and its fringe region, the bed body inside phenomenon of backflowing is suppressed, and makes the total number of theoretical plate of expanded bed when absorption improve greatly.

In the application's the technical scheme, for the expanded bed body construction, fully take into account feed liquid and spray flow velocity (circular flow adds and sees through stream), the seeing through resistance this three influence the interior uniform principal element of fluid pressure of distributor greatly of the size of circulatory flow and sieve plate, adopt uniform fluid distribution design under the low pressure, variable section runner, circulation of distributor inner fluid and the circulation of distributor outer fluid combine, not only can cleaning porous supporting screening plate, simultaneously can carry out uniform to the material liquid that enters expanded bed better, successfully solved expanded bed in the industry's enlarging production process under the low bed layer pressure drop feed liquid liquid supply pressure along the equally distributed problem of radial direction, guaranteed preferably to present the laminar flow state by the fluid of expanded bed bed, can improve/improve the result of use/service efficiency of expanded bed greatly, improve the absorption carrying capacity and the desorption resolution ratio of expanded bed.

Because solid sorbent particles is in suspended state in the expanded bed, the voidage of bed is much larger than the voidage of fixed bed, can make solid particle polluters such as cell in the material liquid, cell fragment smoothly by bed, be dissolved in target composition in the material liquid and be inflated in the bed solid absorbent and catch.Therefore, expanded bed need not the solid particle of material liquid is handled especially, with original centrifugal, filter, catch three steps and synthesize a step, directly carry out chromatogram and catch.For this reason, expanded bed can be applied to replace traditional fixed bed chromatography column in the chromatographic separation process, and shortened process, biochemical operating cost and the production cost that separates reduced.

Among Fig. 6, provided the equipment connection structure schematic relationships of chromatographic isolation unit in the technical program, its expanded bed adopts aforesaid upper nozzle assembly with recoil cleaning function all to press distributor with the liquid with self-cleaning function, concrete, top at the expansion column, be provided with on the feed liquid to go out to manage and the solid dirt delivery pipe,, comb under feed pipe and the feed liquid be set in the bottom of expansion column; In addition, aforesaid porous stops that going out pipe, solid dirt delivery pipe and mobile stick harness on sieve plate, expanded bed upper end wall, the feed liquid constitutes the column cap assembly, be arranged on the upper end (existing expanded bed chromatography splitter all has can upper and lower mobile column cap assembly, down with) of expansion column movably.

The technical program is provided with sample liquid circulating tank V101, buffer solution circulating tank V102, unloads post solution circulating tank V103, the adsorbent tank V104 that has centrifugal solid-liquid separator LXFLQ, feed pump B101, dress post pump B102 and online coarse filter ZXCLQ, constitutes a chromatographic isolation unit that is applicable to biochemical extractor gauge modelling production process with expanded bed T101.

Wherein, the outlet of sample circulating tank V101 and buffer solution circulating tank V102, is connected with the feed pipe of expanded bed T101 with valve F013 by feed pump B101 through valve F006 and F007 through respectively; Go out pipe on the feed liquid of expanded bed and be connected with product jar CPG, or be connected with waste water or intermediate storage jar (not shown) through valve F002 from capital outlet pressure control valve F102 through valve F011, capital outlet pressure control valve F102 and valve F001.

Comb is connected with the liquid back pipe of buffer solution circulating tank with valve F004 through circular flow pressure-regulating valve F101 under the feed liquid of expanded bed; Simultaneously, comb is connected with the liquid back pipe of sample circulating tank with online coarse filter ZXCLQ through circular flow pressure-regulating valve F101, valve F003 under the feed liquid of expanded bed, constitutes closed circuit.

Going out on the feed liquid of expanded bed between pipe and the feed pipe, the bypass conduit by valve F010 control is set.

Going out pipe on the feed liquid of expanded bed capital end is connected with the outlet of feed pump through valve F012.

The solid dirt delivery pipe of expanded bed passes through upper inlet pipe, the upper outlet pipe of valve F015, adsorbent tank and unloads post solution circulating tank V103 and valve F009, is connected with the inlet tube of feed pump.

The following outlet of adsorbent tank is connected with the solid dirt delivery pipe of expanded bed with valve F016 through valve F017, dress post pump B102.

The first atmospheric valve F051 is set on comb road under the feed liquid of expanded bed; The second atmospheric valve F052 is set on the feeding pipe of expanded bed; The 3rd atmospheric valve F014 is set on the solid dirt discharge pipe of expanded bed.

In addition, on the outlet of sample jar YPG, cleaning fluid tank QXYG, wash-out flow container XTYG, buffering flow container HCYG and on the inlet tube of product jar CPG, be respectively arranged with the valve of control piper break-make.

The feed pipe of expanded bed is provided with inlet flow rate meter FIT-2, inlet pressure meter PIT-1 and online conductivity meter Cond, go out pipe on the feed liquid of expanded bed and be provided with online conductivity meter Cond, pH meter PH, rate of discharge meter FIT-1 and ultraviolet tester UV, on the connecting line between expanded bed and the sample circulating tank/buffer solution circulating tank return duct, circulating fluid pressure meter PIT-2 is set.

When expanded bed was in normal operating conditions, the raw material feed liquid in the sample circulating tank was sent into through feed pump and is carried out chromatographic isolation in the expanded bed, and the feed liquid after the separation is exported through the feed liquid efferent duct; The raw material feed liquid of circulating reflux is returned the sample circulating tank again after online coarse filter filters.

When expanded bed was in " recoil clean " state, backwash liquid was pumped into the top of expanded bed through the dress post, and the porous that is positioned at the expanded bed upper end is stopped the sieve plate cleaning that recoils.

Simultaneously, the mobile stick harness in the solid dirt delivery pipe moves down, and opens top jet nozzle outlet, takes the solid particle that gathers under the porous screen cloth out of expanded bed and discharges from the solid dirt delivery pipe.

Simultaneously, utilize mobile stick harness in the upper nozzle assembly on move or move down, close or open the solid dirt delivery pipe, under the cooperation of unloading post solution circulating tank V103, the adsorbent tank V104 that has whizzer LXFLQ, feed pump and dress post pump B102, realize the filling on the throne and/or the unloading of solid absorption particle in the expanded bed.

Further, the sample circulating tank is connected with sample jar, cleaning fluid pipe and/or buffering flow container, and the liquid in sample jar, cleaning fluid pipe and/or the buffering flow container adopts the mode of liquid level control to mend in the sample circulating tank.

Simultaneously, the buffer solution circulating tank is connected with the buffering flow container, and is same, and the liquid in the buffering flow container adopts the mode of liquid level control to mend in the buffer solution circulating tank.

The charging pump intake piping also is connected with the outlet of wash-out flow container through valve F008.

At the outlet of feed pump, be provided for catching and removing the air trap KQXJ of bubble.

Among Fig. 7, by stop in expanded bed column bottom porous sieve plate below the liquid with self-cleaning function be set all press distributor, the upper nozzle assembly that has the recoil cleaning function in the setting of expanded bed capital end, and after adopting equipment connecting relation shown in Figure 6, the chromatographic isolation technological process of the technical program expanded bed chromatography splitter comprises the following steps (this technology is carried out in the scope of room temperature at 4 ℃ usually) at least:

A, chromatographic column balance:

This is the first step of expanded bed chromatography operation.

Feed pump B101 squeezes into buffer solution from buffer solution circulating tank V102 the feed pipe of expanded bed T101 bottom, buffer solution goes out the pipe and the first atmospheric valve F051 emptying earlier on feed liquid, switch to buffer solution circulating tank V102 again, to form suitable circular flow and to cross the post flow.

Fresh buffer constantly mends buffer solution circulating tank V102 in the mode of liquid level control, and the purpose of this operation is to form certain circular flow and cross the post flow.

Crossing post flow and circular flow can be by the feedback of expanded bed bottom inlet flowmeter FIT-1 and expanded bed top exit flowmeter FIT-2, and by control feed pump B101 speed, circular flow pressure-regulating valve F101 and capital outlet pressure control valve F102 realize.

Crossing the post flow is that the expanded bed separating technology is required, and circular flow is for preventing that sieve plate from blocking and guaranteeing that uniform pressure distribution is required.In great majority operations, this circular flow and cross the ratio of post flow can be between 0 to 10, depend on the needs that prevent to block with uniform pressure distribution.By selecting appropriate ratio, can reach the expanded bed of uniform pressure distribution and the high number of theoretical plate of low thermal expansion.

Sample on B, the chromatographic column;

During sample introduction, feed pump B101 import switches to sample liquid circulating tank outlet, and comb switches to the inlet tube (can a first emptying part) of sample liquid circulating tank V101 under the feed liquid of loop exit expanded bed bottom.

Continuation is by inlet flow rate meter and rate of discharge meter FIT-1 and FIT-2 feedback and regulate feed pump B101, circular flow pressure-regulating valve F101 and capital outlet pressure control valve F102, controls required post flow and circular flow excessively.

The sample feed liquid is squeezed into expanded bed adsorption post T101 inlet tube through feed pump B101 among the sample circulating tank V101.

Wherein Yi Bufen feed liquid by comb circulating reflux under the feed liquid of expanded bed bottom to sample circulating tank V101, another part feed liquid is passed expanded bed bottom porous supporting screening plate and is entered expanding bed, by the absorbent particles in the expanded bed target component in the feed liquid is adsorbed.

The feed liquid that has been adsorbed the target component is passed the expanded bed upper porous and is stopped that sieve plate flows out expanded bed, again discharge system or enter the intermediate storage jar after ultraviolet tester, pH meter or online conductivity meter detect.

Fresh sample liquid is constantly mended to the sample circulating tank in the mode of liquid level control.

During the last sample, if obvious rising (the expanded bed operating pressure is usually below 1.5bar) falls in expanded bed pressure, can suspend and go up the sample operation, feed pump B101 import is switched to buffer solution circulating tank V102 (automatic control of liquid level), close the expanded bed bottom inlet, open bypass valve F010, buffer solution is introduced the expanded bed top, the porous at backwash top stops sieve plate; Open top jet nozzle outlet simultaneously, porous is stopped that the solid material that gathers under the sieve plate is taken expanded bed out of and from the emptying of solid dirt delivery pipe.

Recoil is closed feed pump B101 after cleaning and finishing, and closes top jet nozzle, closes top bypass valve F010, opens expanded bed bottom feed pipe; Come back to above-mentioned " going up sample " operation again.This operation can repeatedly be carried out as required.

Intact etc. sample treatment, " going up sample " promptly comes to an end.Usually can establish air horn on the pipeline finishes to show " going up sample ".

If necessary, " go up sample " later stage can add buffer solution to reduce viscosity of sludge and granule density in sample circulating tank V101.

C, chromatographic column flushing;

Behind the end of the sample, feed pump B101 import switches to buffer solution circulating tank V102, and the expanded bed circulation port switches to waste liquid emptying earlier, switches back to buffer solution circulating tank V102 again.

Control same post flow and circular flow excessively, allow expanded bed pass through a certain amount of buffer solution, make ultraviolet tester, pH meter or online conductivity meter instrumentation be stabilized to the technology specified scope, promptly finish the chromatographic column flushing according to technological requirement.

D, adsorbate wash-out;

At first stop feed pump B101, allow the expanded bed bed leave standstill; Again described column cap assembly is reduced to bed and leave standstill interface and fixing, make chromatographic column become the fixed-bed structure state by the expanded bed configuration state.

Then, restart feed pump B101, its import is cut to buffer solution circulating tank V102, and expanded bed chromatography splitter circulation port is closed, and crosses the post flow by inlet flow rate meter FIT-1 FEEDBACK CONTROL.

Allow expanded bed pass through a certain amount of buffer solution according to technological requirement, make ultraviolet tester, pH meter or online conductivity meter instrumentation be stabilized to the technology specified scope.

During wash-out, feed pump B101 import switches to the outlet of wash-out flow container and is connected, and the expanded bed loop exit is still closed, and continues to cross the post flow by inlet flow rate meter FIT-1 FEEDBACK CONTROL.

Monitoring ultraviolet tester, pH meter and/or online conductivity meter, machine switches target peak (product) introducing product jar when appropriate, and all the other put into waste liquid or middle remaining tank.

E, chromatographic column are cleaned and regeneration;

After wash-out is finished, stop feed pump B101, the top porous is stopped the upper nozzle assembly of sieve plate is opened, and by the logical atmosphere of the 3rd atmospheric valve F014.

Then aforesaid column cap assembly is risen to original height, fixedly the column cap assembly makes chromatographic column revert to the expanded bed configuration state, moves on the mobile stick harness, closes the upper nozzle assembly.

With the whole emptying of liquid in the sample circulating tank V101, cleaning fluid self-cleaning flow container QXYG is introduced the sample circulating tank; Start feed pump, feed pump B101 import is switched to sample circulating tank V101, the chromatographic column circulation port switches counter sample product circulating tank V101 (being that circular flow pressure-regulating valve F10 opens), and cleaning fluid is introduced sample circulating tank V101.

According to technological requirement, control required post flow and the circular flow crossed, allow expanded bed pass through a certain amount of cleaning fluid, make ultraviolet tester, pH meter or online conductivity meter instrumentation be stabilized to the technology specified scope, promptly finish the cleaning and the regeneration of chromatographic column.

F, the heavy balance of chromatographic column;

After chromatographic column cleaning and regeneration are finished, close feed pump B101.

With the whole emptying of liquid in the sample circulating tank V101, close each emptying valve, buffer solution is introduced sample circulating tank (by liquid level control).

Start feed pump B101, its import is switched to sample circulating tank V101; The expanded bed circulation port is earlier from solid dirt delivery pipe emptying switchback sample circulating tank V101 again.

According to technological requirement, with required post flow and the circular flow crossed of aforementioned same method control;

Allow chromatographic column pass through a certain amount of buffer solution, make ultraviolet tester, pH meter or online conductivity meter instrumentation be stabilized to the technology specified scope, promptly finish the heavy balance of chromatographic column.

Further, the chromatographic isolation technological process in the technical program also comprises the following steps:

G, dress post:

At first, earlier expanded bed chromatography splitter T101 is filled with buffer solution, again the upper nozzle assembly outlet at expanded bed top is opened;

Start dress post pump B102, absorbent particles suspension (by stirring) is squeezed into the expanded bed T101 from adsorbent tank V104;

The adsorption particle natural subsidence is in the expanded bed chromatography post, and liquid stops that through porous sieve plate brings out a mouthful outflow from the expanded bed capital, realizes dress post on the throne.

H, unload post:

At first, go out pipe on the feed liquid at expanded bed top and close, the top is moved stick harness and is moved down, and the feed pump import switches to unloads post buffer solution circulating tank.

Start feed pump B101 and suitably open big flow, make the fluid velocity in the expanded bed surpass the adsorption particle sinking speed, absorbent particles can be brought into adsorbent tank V104;

Wherein, the centrifugal solid-liquid separator among the adsorbent tank V104 is stayed the adsorption particle medium in the jar, and solution then overflows and recycles.

Further, in " going up sample " the step later stage, can to reduce the viscosity and the granule density of raw material feed liquid, improve the rate of recovery by in the sample circulating tank, injecting dilution.

Further, aforesaid circular flow and the ratio of crossing the post flow are between 0 to 10, and its ratio size depends on the needs that prevent to block with uniform pressure distribution.

In Fig. 8, be " balance " step of expanded bed chromatography exchange column (a), go out on advancing under the raw material feed liquid, and circular flow is arranged in the expanded bed bottom, spray the needs of flow velocity, the uniform effect of feed liquid fluid pressure and self-cleaning function to satisfy feed liquid.

The switching of the annexation of equipment, each valve and flow direction in this step, the existing introduction no longer repeated in aforementioned technological process, down together.

In (b), " going up sample " step of expression expanded bed, need carry out " recoil clean " if obvious rising falls in expanded bed chromatography splitter pressure this moment.

In (c), in expanded bed " recoil is cleaned " step, backwash liquid goes out pipe from feed liquid and enters expanded bed, and porous is stopped that sieve plate carries out the backwash backwash liquid and discharges from the solid dirt delivery pipe.

In (d), expanded bed carries out " chromatographic column flushing " step after " going up sample " step finishes.

In (e), expanded bed carries out " adsorbate wash-out " step.

In (f), expanded bed carries out " cleaning and regeneration " step.

In (g), after chromatographic column cleaning and regeneration are finished, carry out " heavy balance " step.

Dotted region is taken up space by adsorption particle among the figure, in (e) step, clearly because the moving down of aforementioned column cap assembly, makes minimum that adsorption particle takes up space, and actual exchange column device at this moment is to be in the fixed-bed structure state.

In Fig. 9,9a is depicted as the signal of dress post, earlier the expanded bed chromatography splitter is filled with buffer solution, the upper nozzle assembly outlet at expanded bed top is opened again; Start dress post pump, absorbent particles suspension is squeezed into the chromatographic column from adsorbent tank.

Adsorption particle enters the expanded bed from the upper nozzle assembly (being the solid dirt delivery pipe) at expanded bed top, and liquid goes out the pipe outflow from the feed liquid of expanded bed capital end, has realized " dress post " on the throne.

9b is depicted as and unloads the post signal, fluid velocity in the expanded bed surpasses the adsorption particle sinking speed, absorbent particles is taken out of expanded bed from the upper nozzle assembly (being the solid dirt delivery pipe) at expanded bed top, bring adsorbent tank into, centrifugal solid-liquid separator in the adsorbent tank is stayed the adsorption particle medium in the jar, and solution then overflows and recycles.

Continue the operation feed pump and all be moved in the adsorbent tank, realized " unloading post " on the throne until all adsorption particle media.

The technical program has realized automatic " dress post " and " unloading post " process of expanded bed, because there be not " dismounting " or " moving " of expanded bed cylinder, so be referred to as " on the throne " loading, unloading post smoothly owing to adopted the said equipment structure/composition and connecting line.

Embodiment 1: the application of expanded bed in monoclonal antibody is produced

This albumen is analysed in the extracellular by the CHO animal cell expression, the about 10-15um of Chinese hamster ovary celI size, and in 1,500L-10 cultivates final cell density 1X10 in the 000L cell jar ⁷Cells/ml, titre 500mg/ml.

Traditional handicraft needs the high-speed and continuous heart of wandering about as a refugee to add deep layer and filter and to add the albumin A affinity chromatography again, and then uses ion-exchange, and is hydrophobic, or the high-resolution gel chromatography is made into purifying.

Above-mentioned technology needs 2～3 days time to finish affine three steps of look of continuous flow centrifugation, in-depth filtration and albumin A usually.(fixed bed～120-180cm/hr), linear speed moves (height of bed 30cm, 10 minutes time of staying) to affinity chromatography, and carrying capacity is about 20g/L at 180cm/hr.

Chromatographic column volume 37.5L, the about 40cm of diameter.Chromatographic system operation flow is about 225L/hr, and the 1500L sample is finished sample about 6.6hr.

The continuous flow centrifugation apparatus expensive, the in-depth filtration consumable quantity is big, and fixed bed affinity chromatography flux is low, and the last sample time is long.And in-depth filtration can cause cell rupture to discharge other foreign proteins and hydrolase.Can influence product yield as untimely removal.Three step yields are generally 95%, 90% and 97%.Total recovery is 82%.

Adopt the expanded bed technology cellular expression liquid can be directly used in the albumin A affinity chromatography, thereby the three traditional steps are combined into a step.

Adsorbent medium: streamline r-proteinA (GE Healthcare)

Particle diameter :～150um/80-165

Proportion: 1.3g/ml

Mesh size: 20um

The static bed of bed height: 30cm

Linear flow speed: 240cm/hr (200-400cm/hr)

Carrying capacity: 17.5g/L

Column volume: 42L

Column diameter: 50cm (～43cm)

Circular flow/mistake post stream: 2.5

Cross the post flow approximately: 471L/hr

Flow system flow: 1200L/hr

Operating pressure:＜1.0bar

The last sample time: 3.2hr

Yield:＞92%

As seen and traditional handicraft compare, expanded bed technology can be simplified " catching " step greatly, make three the step be simplified to a step, the operation cycle is reduced to 40% (monoclonal antibody albuminoid purifying is even more important) of traditional handicraft approximately.

Simultaneously, because cancelled supercentrifuge and filter plant in the process route, 60% of traditional handicraft is reduced in initial outlay approximately, and it is original 75% that direct material is reduced to approximately, and total recovery also can improve at least 10%.

Embodiment 2: the application of expanded bed in recombinant human albumin (rHSA) is produced

This albumen is analysed in the extracellular by Pichia Pastoris yeast cell to express, in 5, cultivates in the 000L fermentation tank, last titre～1.0g/l.

Traditional technology needs the high-speed and continuous heart of wandering about as a refugee to add deep layer and filter, and add ion-exchange chromatography again, and then with hydrophobic, into purifying is made in ion-exchange.Traditional handicraft needs 2-3 days time to finish continuous flow centrifugation usually, and three steps of chromatogram are caught in in-depth filtration and ion-exchange.(fixed bed～120-180cm/hr) linear speed operation (height of bed 30cm, 15 minutes time of staying), carrying capacity is about 10g/L at 120cm/hr in ion-exchange.Chromatographic column volume 500L, the about 140cm of diameter (145cm).Chromatographic system operation flow is about 1900L/hr, and the 5000L sample is finished sample about 2.6hr.The continuous flow centrifugation apparatus expensive, the in-depth filtration consumable quantity is big.And in-depth filtration can cause cell rupture to discharge other foreign proteins and hydrolase.Can influence product yield as untimely removal.Three step yields are generally 90%, 90% and 95%.Total recovery is 76%.

Adopt the expanded bed technology cellular expression liquid can be directly used in ion-exchange chromatography, thereby the three traditional steps are combined into a step.

Adsorbent medium: streamline SP (GE Healthcare)

Particle diameter :～200um/100-300um

Proportion: 1.2g/ml

Mesh size: 20um

The static bed of bed height: 30cm

Linear flow speed: 250cm/hr 2X (200-400cm/hr)

Carrying capacity: 8.5g/L

Column volume: 588L

Column diameter: 160cm (～158cm)

Circular flow/mistake post stream: 3.0

Cross the post flow approximately: 5000L/hr

Flow system flow: 15000L/hr

Operating pressure:＜1.0bar

The last sample time: 1.0hr

Yield:＞90%

Can learn equally and traditional handicraft is compared that expanded bed technology can be simplified greatly and catches step, makes three to go on foot into a step, the operation cycle reduces to 40% approximately.Because removed centrifuge and filter plant, 60% of traditional handicraft is reduced in initial outlay approximately, and it is original 75% that direct material is reduced to approximately, and total recovery can improve 30%.

Embodiment 3: the application of expanded bed in antibiotics production

The antibiotics production scale is big, and the flow height is another good field of using the expanded bed technology.

Along with medical industry is more and more higher to antibiotic purity requirement, ion-exchange chromatography becomes the essential means in the downstream separation technology gradually, especially the situation that contains cotton-shaped solid in the zymotic fluid and can't use filtration once to remove, the expanded bed The Application of Technology can have remarkable economic efficiency.

Embodiment 4: the application of expanded bed in protein isolate from milk (Lactoferrin) is produced

Because handle the ability of high viscosity and solid impurity, the expanded bed technology is being that significant advantage and tangible economic benefit are also arranged aspect the transgene protein of basal expression with milk.

To sum up, the technical program is even more important in following field: monoclonal antibody (dosage is big, common annual production tonne), recombinant human albumen (surpasses 500 tons of/year whole world, tens tonnes of common annual productions), transgenosis milk expressed proteins medicine etc. (material viscosity height, foreign protein is many).

In the technical program, by stop in expanded bed bottom porous sieve plate below the upper nozzle assembly that the liquid with self-cleaning function is all pressed distributor and had the recoil cleaning function in the setting of expanded bed capital end is set, make the porous at expanded bed top stop that the porous screen cloth gripper shoe of sieve plate and/or expanded bed bottom can have backwash and/or self-cleaning function, make the expanded bed chromatography splitter be applicable to and handle the raw material feed liquid that contains solid particle polluter, solved the difficult problem in solid particle obstruction sieve plate and duct.

Simultaneously, because in the application's technical scheme, fluid course size and porous screen cloth/sieve plate resistance determined when design, circular flow and to see through flow then be the controllable operating variable, so the whole control of its expanded bed is convenient, the industrially scalable amplification is easy; Simultaneously, dead volume is little in the distributor, the mixed bed of distributor internal return is compared and can be ignored, thereby can realize the uniform fluid distribution in the small size under the low pressure, make the industrial applications of expanded bed on biochemical isolation technics of low thermal expansion, high number of theoretical plate be achieved, reach the core advantage of using expanded bed.

After adopting the application's technical scheme, the expanded bed technology can be applicable to the purifies and separates of the protein matter of useful microorganism of institute and zooblast production, allows to contain solid particle or sedimentary material is directly used in chromatographic isolation.

Can learn that in conjunction with foregoing the main innovate point of the application's technical scheme is:

1) proposed a kind of expanded bed splitter of new structure, it can realize high flux, high number of theoretical plate chromatographic isolation to containing solid cell liquid in the biological industry or lysate is directly handled; Simplified biochemical separation processes flow process, especially to monoclonal antibodies, human body recombinant albumin etc. needs the product of large-scale production significant.

2) bottom of the porous screen cloth gripper shoe of expanded bed adsorption post is provided with a liquid distribution trough in the technical program, and this distributor has uniform ability of better liquid and self-cleaning function; This uniform device has low pressure drop, small size, advantages such as self-stabilization.

3) expanded bed top is provided with the recoil inlet of buffer solution in the technical program, can remove the solid particle accumulation of expanded bed upper porous sieve plate bottom and stops up.

4) in the technological process at above-mentioned improvement expansion of structure bed, in sample raw material circulation stream, filter is set, can removes the agglomerate solid dirt in the circulation sample feed liquid, guarantee that new expanded bed technological process carries out smoothly.

5) can realize dress post on the throne and unload post, liquid-solid cyclone separator is set in resin container, when the resin of unloading expanded bed, be convenient to separating of polymeric adsorbent and buffer solution, and buffer solution can be recycled.

6) the direct processing of pair cell lysate is removed from centrifugal and filtration step, and buying expenses of equipment reduces obviously.

7) can realize whole process cleaning on the throne, dress post on the throne and unload post, help realizing the full-automation control of whole process of production.

The application's technical scheme is particularly suitable for realizing the chromatographic isolation of high flux, high number of theoretical plate to containing solid cell liquid in the biological industry or cell pyrolysis liquid is directly handled; It has removed existing " centrifugal " and " filtration " processing step from, has simplified the technological process of production of biochemical separation, and especially to monoclonal antibodies, human body recombinant albumin etc. needs the product of large-scale industrial production significant, and economic benefit is obvious.

Claims

1. an expanded bed chromatography splitter that is used for biochemical separation processes comprises that the porous that is positioned at expanded bed capital end stops sieve plate and discharge nozzle, is positioned at the porous supporting screening plate and the feed pipe of expansion column bottom, it is characterized in that:

On the top of expanded bed, be provided with and comprise and go out pipe, solid dirt delivery pipe, mobile stick harness and sealing ring on the feed liquid the having upper nozzle assembly of the cleaning function that recoils at least;

In the bottom of described expanded bed, be provided with and comprise feed pipe, radial spray hole, comb under water conservancy diversion disk and the feed liquid radially at least, the liquid with self-cleaning function is all pressed distributor;

When porous stops the sieve plate below solid particle accumulation is arranged, cause porous to stop when the resistance drop of sieve plate increases, backwash liquid goes out pipe from feed liquid and enters expanded bed capital end, porous is stopped the sieve plate cleaning that recoils, blow off accumulating in the solid accumulation that porous stops the sieve plate bottom, move down mobile stick harness simultaneously, open the solid dirt delivery pipe, the solid accumulation is discharged from the solid dirt delivery pipe; After recoil finishes, on move mobile plunger, close the solid dirt delivery pipe, expanded bed is proceeded the operation of normal chromatographic isolation;

Described radially water conservancy diversion disk cooperates with the lower surface of porous screen cloth gripper shoe, on the radial direction of water conservancy diversion disk, constitutes variable cross-section uniform fluid distribution runner at the center of water conservancy diversion disk radially; The raw material feed liquid that from described radial spray hole, ejects, along the expanded bed or the radial direction of water conservancy diversion disk radially, to spraying all around, under the guiding of water conservancy diversion disk radially, the porous supporting screening plate is carried out side direction/tangentially wash away, the solid particle flushing reflux main flow that will assemble in the sieve plate bottom, comb is discharged under the feed liquid, turn back in the sample circulating tank and circulate, realize cleaning complete on the throne and automatic cleaning action the porous supporting screening plate;

The upper nozzle assembly and the liquid that have recoil cleaning and/or self-cleaning function by setting are all pressed distributor, make the expanded bed chromatography splitter be applicable to and handle the raw material feed liquid that contains solid particle polluter, improve/improve the result of use/service efficiency of expanded bed in the industrial-scale production of biochemical field with this.

2. the expanded bed chromatography splitter that is used for biochemical separation processes as claimed in claim 1, it is characterized in that on the described feed liquid going out that pipe and solid dirt delivery pipe are arranged on the expanded bed wall that porous stops the sieve plate top, described solid dirt delivery pipe runs through the middle part setting that expanded bed upper end wall and porous stop sieve plate; Described mobile stick harness and sealing ring are arranged in the solid dirt delivery pipe;

When moving on the described mobile stick harness, it cooperates with sealing ring, closes the solid dirt delivery pipe, and described upper nozzle assembly is in normal operating conditions, and the raw material feed liquid in the expanded bed is flowed through after porous stops sieve plate, discharges by going out pipe on the feed liquid; When described mobile stick harness moves down, the conducting of solid dirt delivery pipe, described upper nozzle assembly are in recoil cleaning state, and backwash liquid is through going out to manage reverse flow after porous stops sieve plate on the feed liquid, to accumulate in porous and stop that sieve plate bottom solid accumulation blows off, and is discharged by the solid dirt delivery pipe; After the recoil cleaning finishes, move on the mobile stick harness, close the solid dirt delivery pipe, can proceed normal chromatographic isolation operation;

Simultaneously, utilize mobile stick harness in the upper nozzle assembly on move or move down, close or open the solid dirt delivery pipe, be used for realizing the filling on the throne and/or the unloading of solid absorption particle in the expanded bed.

3. the expanded bed chromatography splitter that is used for biochemical separation processes as claimed in claim 1, it is characterized in that feed pipe being set in the centre of described expansion column bottom, the top of feed pipe is arranged on the below of porous supporting screening plate, on the top of described feed pipe the radial spray hole is set; Between expansion column bottom wall and porous supporting screening plate, radially water conservancy diversion disk is set; Offer through hole in the centre of water conservancy diversion disk radially, described feed pipe runs through the radially through hole setting of water conservancy diversion disk; Described radially water conservancy diversion disk cooperates with the lower surface of porous supporting screening plate, on the radial direction of water conservancy diversion disk, constitutes variable cross-section uniform fluid distribution runner at the center of water conservancy diversion disk radially; Comb is arranged on the expanded bed wall of porous supporting screening plate below under the described feed liquid;

4. the expanded bed chromatography splitter that is used for biochemical separation processes as claimed in claim 1 is characterized in that the upper surface at described radially water conservancy diversion disk, is provided with the deflector that polylith is erect along radial direction, and described deflector is pressed the uniform arrangement setting of circumferencial direction; Below the described radially water conservancy diversion disk and between the wall of expanded bed lower end, the runner that backflows is set; The diameter of described radially water conservancy diversion disc centre position through hole between through hole and feed pipe outer tube wall, forms a liquid communication gap more than or equal to the external diameter of described feed pipe, constitutes the internal liquid circulatory flow.

5. expanded bed chromatography splitter that is used for biochemical separation processes as claimed in claim 1, comprise the expansion column, top at described expansion column, be provided with and go out pipe and solid dirt delivery pipe on the feed liquid, in the bottom of described expansion column, comb under feed pipe and the feed liquid is set, described porous stops that going out pipe, solid dirt delivery pipe and mobile stick harness on sieve plate, expanded bed upper end wall, the feed liquid constitutes the column cap assembly, be arranged on the upper end of expansion column movably, it is characterized in that:

Sample liquid circulating tank, buffer solution circulating tank are set, unload post solution circulating tank, the adsorbent tank that has the centrifugal solid-liquid separator, feed pump, dress post pump and online coarse filter, constitute a chromatographic isolation unit that is applicable to biochemical extractor gauge modelling production process with described expansion column;

Wherein, the outlet of described sample circulating tank and buffer solution circulating tank is connected with the feed pipe of expanded bed through feed pump; Go out pipe on the feed liquid of described expanded bed and be connected with the product jar, perhaps, go out pipe on the feed liquid and be connected with waste water or intermediate storage jar through capital outlet pressure control valve through capital outlet pressure control valve;

Comb is connected with the liquid back pipe of online coarse filter with the sample circulating tank through the circular flow pressure-regulating valve under the feed liquid of described expanded bed, constitutes closed circuit;

Comb is connected through the liquid back pipe of circular flow pressure-regulating valve with the buffer solution circulating tank under the feed liquid of described expanded bed;

Going out on the feed liquid of described expanded bed under pipe and the feed liquid between the comb, bypass conduit is set;

Going out pipe on the feed liquid of described expanded bed capital end is connected with the outlet of feed pump through valve;

The solid dirt delivery pipe of described expanded bed passes through upper inlet pipe, the upper outlet pipe of adsorbent tank and unloads post solution circulating tank, is connected with the inlet tube of feed pump;

The following outlet of described adsorbent tank is connected with the solid dirt delivery pipe of expanded bed through dress post pump;

First atmospheric valve is set on comb road under the feed liquid of described expanded bed;

Second atmospheric valve is set on the feeding pipe of described expanded bed;

On the solid dirt discharge pipe of described expanded bed, the 3rd atmospheric valve is set;

On the pipe of the import/export of described expanded bed or each jar, valve is set;

Inlet flow rate meter, inlet pressure meter and online conductivity meter are set on the feed pipe of expanded bed, on the feed liquid of expanded bed, go out to be provided with on the pipe online conductivity meter, pH meter, rate of discharge meter and ultraviolet tester, on the connecting line between expanded bed and the sample circulating tank/buffer solution circulating tank return duct, the circulating fluid pressure meter is set;

When described expanded bed was in normal operating conditions, the raw material feed liquid in the sample circulating tank was sent into through feed pump and is carried out chromatographic isolation in the expanded bed, and the feed liquid after the separation goes out pipe output on feed liquid; The raw material feed liquid of comb circulating reflux is returned the sample circulating tank again after online coarse filter filters under the feed liquid;

When described expanded bed was in recoil cleaning state, backwash liquid was pumped into the top of expanded bed through the dress post, and the porous that is positioned at the expanded bed upper end is stopped the sieve plate cleaning that recoils; Simultaneously, the mobile stick harness in the solid dirt delivery pipe moves down, and opens top jet nozzle outlet, takes the solid particle that gathers under the porous screen cloth out of expanded bed and discharges from the solid dirt delivery pipe;

After the recoil cleaning finishes, move on the mobile stick harness in the solid dirt delivery pipe, close the solid dirt delivery pipe, proceed normal chromatographic isolation operation;

6. the expanded bed chromatography splitter that is used for biochemical separation processes as claimed in claim 5, it is characterized in that described sample circulating tank is connected with sample jar, cleaning fluid tank and/or buffering flow container, the liquid in described sample jar, cleaning fluid pipe and/or the buffering flow container adopts the mode of liquid level control to mend in the sample circulating tank;

Described buffer solution circulating tank is connected with the buffering flow container, and the liquid in the described buffering flow container adopts the mode of liquid level control to mend in the buffer solution circulating tank;

Described charging pump intake piping is connected with the outlet of wash-out flow container;

7. one kind as being used for the chromatographic isolation technology of the expanded bed chromatography splitter of biochemical separation processes as described in the claim 5, it is characterized in that:

The chromatographic isolation technological process of described expanded bed chromatography splitter comprises the following steps: at least

A, chromatographic column balance:

After flow pressure is stable, allow chromatographic column pass through a certain amount of buffer solution, make ultraviolet tester, pH meter or online conductivity meter instrumentation be stabilized to the technology specified scope, promptly finish the chromatographic column balance according to technological requirement;

Sample on B, the chromatographic column;

Intact etc. sample treatment, last sample promptly comes to an end;

C, chromatographic column flushing;

Behind the end of the sample, the feed pump import switches to the buffer solution circulating tank, and the expanded bed circulation port switches to waste liquid emptying earlier, switches back to the buffer solution circulating tank again; Control same post flow and circular flow excessively, allow chromatographic column pass through a certain amount of buffer solution, make ultraviolet tester, pH meter or online conductivity meter instrumentation be stabilized to the technology specified scope and promptly finish the chromatographic column flushing according to technological requirement;

D, adsorbate wash-out;

Monitoring ultraviolet tester, pH meter and/or online conductivity meter, the machine switching is introduced the product jar with target peak when appropriate, and all the other put into waste liquid or middle remaining tank;

E, chromatographic column are cleaned and regeneration;

According to technological requirement, control required post flow and the circular flow crossed.Allow chromatography column pass through a certain amount of cleaning fluid, make ultraviolet tester, pH meter or online conductivity meter instrumentation be stabilized to the technology specified scope, promptly finish the cleaning and the regeneration of chromatographic column;

F, the heavy balance of chromatographic column;

8. as being used for the chromatographic isolation technology of the expanded bed chromatography splitter of biochemical separation processes as described in the claim 7, it is characterized in that described chromatographic isolation technological process also comprises the following steps:

G, dress post:

The adsorption particle natural subsidence is in chromatographic column, and liquid stops that through porous sieve plate brings out a mouthful outflow from the expanded bed capital, realizes dress post on the throne;

H, unload post:

The feed pump import switches to unloads post buffer solution circulating tank;

9. as being used for the chromatographic isolation technology of the expanded bed chromatography splitter of biochemical separation processes as described in the claim 7, it is characterized in that in described " going up sample " the step later stage, can to reduce the viscosity and the granule density of raw material feed liquid, improve the rate of recovery by in the sample circulating tank, injecting dilution.

10. as being used for the chromatographic isolation technology of the expanded bed chromatography splitter of biochemical separation processes as described in the claim 7, the ratio that it is characterized in that described circular flow and cross the post flow depends on the needs that prevent to block with uniform pressure distribution between 0 to 10.