CN105107230B - A kind of chromatograph reequips the method for perfusion culture purified - Google Patents
A kind of chromatograph reequips the method for perfusion culture purified Download PDFInfo
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- CN105107230B CN105107230B CN201510568307.6A CN201510568307A CN105107230B CN 105107230 B CN105107230 B CN 105107230B CN 201510568307 A CN201510568307 A CN 201510568307A CN 105107230 B CN105107230 B CN 105107230B
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Abstract
The present invention relates to chromatograph to be reequiped the method for perfusion culture purified, including：(1) it is adapted as that there are two streams to run two instruments of pillar simultaneously parallel on the basis of chromatograph, a stream and pillar are used for sample loading, and another stream and pillar are balanced wash-out simultaneously；(2) pillar is added on the basis of (1), two streams are parallel, and three pillars run two-by-two；(3) it is adapted as that there are two streams to run three instruments of pillar simultaneously parallel on the basis of chromatograph, one stream and pillar are used for sample loading, another stream and pillar are balanced wash-out simultaneously, and the 3rd pillar can receive the efflux from other two pillars；(4) two parallel refiting schemes of stream with amplification versatility.Present invention repacking is easy, easy to operate, and process is simple, perfusion culture downstream process is carried out continuously and is realized automatic operating, and can realize lab scale to the parallel amplification of production, is easy to large-scale industrial production.
Description
Technical field
The invention belongs to isolate and purify field and perfusion culture technique field, more particularly to a kind of chromatograph is reequiped for filling
The method for flowing culture purified.
Background technology
Traditional fed-batch cell culture usually using thousands of to up to ten thousand liters retort, incubation time be 7-21 days,
The product yield scope of different antibodies is 2-6g/L.And perfusion culture technique continuously adds fresh cultured by reactor
Base, while taking out the culture medium exhausted, can maintain the longer cell culture time, reach as high as the several months.
The advantage of perfusion culture process is that can obtain cell density higher, and in same size reactor water
It is flat, more Product yields can be obtained.Generally, the operation concentration of perfusion culture is 10-30 times of fed-batch culture,
For example, using the bioreactor of 50L, adapted perfusion culture technique, its yield can reach 1,000L fed-batch biological respinses
The production capacity of device.
Additionally, perfusion culture technique also has a clear superiority in terms of product quality, in cell perfusion incubation, give up
Liquid will not be accumulated, because the culture medium exhausted continuously is replaced and removed by fresh culture.The results of product and purifying are also faster
Speed, this is particularly advantageous for the production of unstable product (such as VIII factors).With the progress of technique and to continuous culture gradually
Understand, in 10 to 15 years futures, continuous culture is possible will the main flow production platform as " factory of the future ".
But perfusion cultures are also subject to certain restrictions, such as substantial amounts of harvest liquid is continuously accumulated, and increases downstream processes amount.
The stream of general chromatograph only has one, and systems pumps and sample pump are connected in seven port valves, then with flow to valve and be connected, then connect two
Post position valve, is finally connected with the detector and collector of system.This kind of stream can only run the solution of single pillar, i.e. sample pump
There can only be one kind to enter pillar with the solution of systems pumps.This chromatograph is directed to batch production design, it is impossible to connected
Subsequent layers are analysed.The problem design repacking chromatograph that the present invention is continuously accumulated for harvest liquid, is capable of achieving the continuous processing to harvest liquid,
Directly the continuous capture purpose product from harvest liquid, while can realize that filler full load condition is run, greatly reduces consolidating for downstream
Determine cost and human cost.
The content of the invention
The technical problems to be solved by the invention are to provide a kind of chromatograph and reequip method for perfusion culture purified, solution
Certainly chromatograph is unable to the problem of continuous processing to harvest liquid, reduces the fixed cost and human cost in downstream, and with two streams
Run many functions of pillar simultaneously parallel in road.
The technical solution adopted for the present invention to solve the technical problems is：A kind of chromatograph is provided to reequip for perfusion culture
The method of purifying, wherein described method is：Two valves are used on chromatograph, when described two valves are seven port valves, by the
5. number mouth of one seven port valve and 7. number mouth respectively with second 3. number mouth of seven port valves, 1. a number mouth is connected, described second
A 5. number mouth for seven port valves is shut, and the 7. number mouth of second described seven port valve is products export, is connected to the detector of system instrument
On, the 4. number mouths of two described seven port valves is waste liquid port, and two pillars are attached separately to the 2. and 6. number mouth of two seven port valves
Between, systems pumps solution is entered by the 1. number mouth of first seven port valve, and sample pump solution is entered by the 3. number mouth of first valve, is led to
The valve position state for crossing two seven port valves of switching realizes that two streams run two pillars simultaneously parallel, and a stream and pillar are used for
Sample loading, another stream and pillar are balanced wash-out simultaneously, and with the function of automatically switching.
When two described valves are seven port valves, first 3. number mouth of seven port valves are connected with a 5. number mouth threeway and are connected to
First upper end of pillar, second 3. number mouth of seven port valves is connected with a 5. number mouth threeway and is connected under first pillar
End, second upper end of pillar is connected to by the 1. number mouth of first seven port valve, and a 1. number mouth for second seven port valve is connected to second
The lower end of pillar, first 4. number mouth of seven port valves is connected the upper end for being connected to the 3rd pillar with 6. number mouth threeway, will
Second 4. number mouth of seven port valves is connected the lower end for being connected to the 3rd pillar with 6. number mouth threeway, and second seven port valve is 2.
Number mouth is waste liquid port, and second 7. number mouth of seven port valves is products export, is connected on the detector of system instrument, systems pumps solution
Entered by the 7. number mouth of first seven port valve, sample pump solution is entered by the 2. number mouth of first seven port valve, is led to by switching seven
The valve position state of valve realizes that two streams are parallel, and three pillars run two-by-two, and a stream and pillar are used for sample loading, another
Bar stream and pillar are balanced wash-out simultaneously, with the function of automatically switching, and change pillar during convenient continuous production.
When two described valves are replaced with seven port valves and three eight ways valves, the 3. number mouth of seven port valves is used with 5. number mouth
Threeway connection is connected to first upper end of pillar, and first lower end of pillar connects first eight ways valve, by seven port valves 1.
Number mouth is connected to second upper end of pillar, second lower termination of pillar, second eight ways valve, by the 4. number mouth of seven port valves with 6.
Number mouth threeway connection is connected to the 3rd upper end of pillar, the 3rd eight ways valve of the 3rd lower termination of pillar, described three
A 2. number outlet for individual eight ways valve is waste liquid port, and the 1. number outlet of three eight ways valves converges into a stream to produce by threeway or four-way
Product outlet, it is connected on the detector of system instrument, the 3. number mouth of first eight ways valve and 4. number mouth are connected to the by threeway respectively
Three pillars and second pillar upper end, the 3. number mouth of second eight ways valve and 4. number mouth is connected to the 3rd post by threeway respectively
Son and first pillar upper end, the 4. 3. number mouth of the 3rd eight ways valve and number mouth are connected to second pillar and the by threeway respectively
A piece pillar upper end, systems pumps solution is entered by the 7. number mouth of seven port valves, and sample pump solution is entered by the 2. number mouth of seven port valves, is led to
The valve position state for crossing seven port valves of switching and eight ways valve realizes that there are two streams simultaneously to run three instruments of pillar, a stream simultaneously
Road and pillar are used for sample loading, and another stream and pillar are balanced wash-out simultaneously, and the 3rd pillar can receive to come from
The efflux of other two pillars, and the circulation that can automatically switch of three pillars carried out, and the condition of yield is not being sacrificed so as to reach
Under significantly cost-effective purpose.
When two described valves are replaced with five eight ways valves, the eight ways valve of post upper end two, the 4th eight ways valve 1., 2.,
3. number mouth respectively with the 5th eight ways valve 3., 1., 2. a number mouth is connected and is connected to first pillar, by threeway
Two pillars and the 3rd pillar upper end, and the lower end of first pillar, second pillar and the 3rd pillar connects first respectively
Individual eight ways valve, second eight ways valve and the 3rd eight ways valve, the 4. 3. number mouth of first eight ways valve and number mouth pass through threeway respectively
It is connected to the 3rd pillar and second pillar upper end, the 3. number mouth of second eight ways valve and 4. number mouth is connected to by threeway respectively
Three pillars and first pillar upper end, the 3. number mouth of the 3rd eight ways valve and 4. number mouth is connected to second post by threeway respectively
Son and first pillar upper end, a 2. number outlet for first, second and the 3rd eight ways valve is waste liquid port, and 1. number mouth passes through three
Logical or four-way converges into a stream and is products export, is connected on the detector of instrument, and systems pumps solution is by the 4th eight ways valve
The heart enters, and sample pump solution is entered by the 5th eight ways valve center, and two are realized by the valve position state for switching five eight ways valves
Stream is parallel, and three pillars run two-by-two, and a stream and pillar are used for sample loading, and another stream and pillar are carried out simultaneously
Balance wash-out, pillar is changed with the function of automatically switching, and during convenient continuous production, or with the parallel same luck of two streams
Three instruments of pillar of row, a stream and pillar are used for sample loading, and another stream and pillar are balanced wash-out simultaneously,
3rd pillar can receive the efflux from other two pillars, and the circulation that can automatically switch of three pillars is carried out, so that
Reach the function of significantly cost-effective purpose under conditions of yield is not sacrificed.
When two described valves are replaced with six seven port valves, wherein the 2. number mouth of three seven port valves is connected to three pillars respectively
Upper end, the 7. number mouth of the port valve of the other three seven is connected to three lower ends of pillar, first seven port valves, second seven port valve respectively
6. number mouth threeway of 4. number mouth and the 4th seven port valves, the 5th seven port valves with the 3rd seven port valve and six seven port valves or
Four-way is connected together, and systems pumps solution is entered by the 1. number mouth of first seven port valve, second seven port valve and the 3rd seven port valves,
Sample pump solution is entered by the 3. number mouth of first seven port valve, second seven port valve and the 3rd seven port valves, described first
Seven port valves, second seven port valve and the 3rd seven port valve 5., 6., 7. a number mouth is shut, described the 4th seven port valves, the 5th
Seven port valves and six 5. number mouths of seven port valves are waste liquid port, 1. number mouth is for products export is connected on the detector of system, 2. number mouth
Shut, the 3. number mouth of the port valve of three after each pillar seven and 4. number mouth short circuit, by switching six valve position states of seven port valves,
Realize that there are two streams to run three instruments of pillar simultaneously parallel, a stream and pillar are used for sample loading, another
Stream and pillar are balanced wash-out simultaneously, and the 3rd pillar can receive the efflux from other two pillars, and three
The pillar circulation that can automatically switch is carried out, so as to reach significantly cost-effective purpose under conditions of yield is not sacrificed.
A kind of chromatograph reequips the method for perfusion culture purified, wherein described method is：On new chromatograph
Using two valves, when two described valves are seven port valves, first pillar and second pillar are connected to two seven port valves respectively
Between LoopE mouthfuls and LoopF mouthfuls, first the Syp of seven port valves mouthfuls is connected with second the Col of seven port valves mouthfuls, and first seven leads to
W2 mouthfuls of valve is connected with second the Sap of seven port valves mouthfuls, and systems pumps solution is entered by first the Col of seven port valves mouthfuls, sample pump
Solution is entered by first the Sap of seven port valves mouthfuls, and the W1 of two seven port valves is waste liquid port, and second the Syp of seven port valves mouthfuls is
Products export, it is connected on the detector of system instrument, by switching two valve position states of seven port valves, realizes that two streams are parallel
Two pillars are run simultaneously, and a stream and pillar are used for sample loading, and another stream and pillar are balanced wash-out simultaneously,
And with the function of automatically switching.
One of them is seven port valves to two described valves, when another is post position valve, LoopE mouthful of described seven port valves with
LoopF mouthfuls of short circuit, first pillar upper end of Col mouthfuls of connection, and W1 mouthful be connected to second pillar upper end with W2 mouthfuls by threeway,
2A mouthfuls and 1B mouthfuls of described post position valve is connected to first pillar lower end by threeway, and the 2B and 1A of post position valve are passed through into threeway
Be connected to second lower end of pillar, systems pumps solution by seven port valves Syp mouthfuls of entrance, sample pump solution by seven port valves Sap
Mouth enters, and the valve import of post position is waste liquid port, and post position valve outlet is products export, is connected on the detector of system instrument, by cutting
The valve position state of seven port valves and the different posts position of post position valve are changed, realizes that two streams run two pillars, a stream simultaneously parallel
Road and pillar are used for sample loading, and another stream and pillar are balanced wash-out simultaneously, and with the function of automatically switching.
One of them is post position valve to two described valves, when another is seven port valve, 3A mouthfuls and 1B of described post position valve
Mouthful by threeway be connected to first pillar upper end, 1A mouthfuls with 2B mouthfuls by threeway be connected to second pillar upper end and 2A mouthfuls
The 3rd pillar upper end is connected to by threeway with 3B mouthfuls, LoopE mouthfuls of described seven port valves is connected to the with LoopF mouthfuls by threeway
Piece pillar lower end, W1 mouthfuls second pillar lower end is connected to by threeway with W2 mouthfuls and Col mouthfuls is connected to the lower end of post 3, systems pumps are molten
Liquid is entered by the import of post position valve, and sample pump solution is entered by the outlet of post position valve, and Sap mouthfuls of seven described port valves is waste liquid
Mouthful, Syp mouthful of described seven port valves be products export, be connected on the detector of system instrument, by the valve for switching seven port valves
The different posts position of state and post position valve, realizes that two streams are parallel, and three pillars run two-by-two, and a stream and pillar are used for sample
Product loading, another stream and pillar are balanced wash-out simultaneously, with the function of automatically switching, and during convenient continuous production more
Change pillar.
Two described valves are two post position valves or two seven port valves, and it connects stream and a post position valve and one seven logical
Valve stream is consistent, by switching the valve position state of two seven port valves or two post position valves, realizes that two streams are parallel, three pillars
Run two-by-two, a stream and pillar are used for sample loading, and another stream and pillar are balanced wash-out simultaneously, with automatic
The function of switching, and change pillar during convenient continuous production.
Beneficial effect
Advantages of the present invention has：
(1) method of the present invention can realize that two streams run two pillars simultaneously parallel, so as to reach technique serialization；
(2) present invention can realize that every pillar is fully loaded with, and significantly cost-effective mesh is reached under conditions of yield is not sacrificed
's.
(3) present invention can be used for the repacking of other purifying instruments, be particularly suitable for large-scale production, can flexible design pillar
Usage quantity.
(4) present invention repacking is easy, easy to operate, and process is simple, and perfusion culture downstream process can be made to be carried out continuously and real
Existing automatic operating
(5) present invention can realize lab scale to the parallel amplification of production for chromatograph repacking, be easy to large-scale industry metaplasia
Produce.
Brief description of the drawings
Fig. 1：Chromatograph-two seven port valves, two refiting schemes of post；
Fig. 2：Chromatograph-two seven port valves, three refiting schemes of post；
Fig. 3：Three refiting schemes of pillar of the port valve of chromatograph-seven and three eight ways valves；
Fig. 4：Three refiting schemes of pillar of-five eight ways valves of chromatograph (or equivalent)；
Fig. 5：New chromatograph-two seven port valves, two refiting schemes of post；
Fig. 6：The port valve of new chromatograph-seven and post position two refiting schemes for post of valve；
Fig. 7：The port valve of new chromatograph-seven and post position three refiting schemes for post of valve；
Fig. 8：Chromatograph-six seven port valves, three refiting schemes of pillar；
Fig. 9 a~Fig. 9 c：Inject, Direct inject, Sample pump of the respectively new port valve of chromatograph seven
The position state diagram of load valves.
Fig. 9 d~Fig. 9 f：The respectively LOAD (Down Flow) of the port valve of chromatograph seven, INJECT (Up Flow), WASTE
Valve position state diagram.
Specific embodiment
With reference to specific embodiment, the present invention is expanded on further.It should be understood that these embodiments are merely to illustrate the present invention
Rather than limitation the scope of the present invention.In addition, it is to be understood that after the content for having read instruction of the present invention, people in the art
Member can make various changes or modification to the present invention, and these equivalent form of values equally fall within the application appended claims and limited
Scope.
Embodiment 1
As shown in Fig. 1, Fig. 9 d and Fig. 9 e, a kind of chromatograph reequips the method for perfusion culture purified, wherein described
Method is：Two valves are used on chromatograph, when two described valves are seven port valve 1,2, by first 5. number mouth of seven port valves 1
7. number mouth is respectively with second 3. number mouth of seven port valves 2,1. a number mouth is connected, the 5. number mouth envelope of described second seven port valve 2
Extremely, the 7. number mouth of second described seven port valve 2 is products export, is connected on the detector of system instrument, described two seven
A 4. number mouth for port valve 1,2 is waste liquid port, and two pillars 3,4 are attached separately to 2. and 6. between number mouth of two seven port valves, system
Pump solution is entered by the 1. number mouth of first seven port valve 1, and sample pump solution is entered by the 3. number mouth of first valve 1, by switching
The valve position state of two seven port valves 1,2 realizes that two streams run two pillars simultaneously parallel, and a stream and pillar are used for sample
Product loading, another stream and pillar are balanced wash-out simultaneously, and with the function of automatically switching.
As shown in Fig. 2 a kind of chromatograph reequips the method for perfusion culture purified, wherein described method is：In layer
Two valves are used in analyzer, when two described valves are seven port valve 1,2, first 3. number mouth of seven port valves 1 is used with 5. number mouth
Threeway connection is connected to first upper end of pillar 3, and second 3. number mouth of seven port valves 2 is connected and is connect with a 5. number mouth threeway
In first lower end of pillar 3, the 1. number mouth of first seven port valve 1 is connected to second upper end of pillar 4, second seven port valve
1. number mouth be connected to second lower end of pillar 4, first 4. number mouth of seven port valves is connected with a 6. number mouth threeway and is connected to
3rd upper end of pillar 5, second 4. number mouth of seven port valves is connected with 6. number mouth threeway and is connected to the 3rd pillar 5
Lower end, second 2. number mouth of seven port valves 2 is waste liquid port, and second 7. number mouth of seven port valves 2 is products export, is connected to system instrument
On the detector of device, systems pumps solution is entered by the 7. number mouth of first seven port valve 1, and sample pump solution is by first seven port valve 1
2. number mouth enter, realize that two streams are parallel by the valve position states for switching two seven port valves 1,2, three pillars are transported two-by-two
OK, a stream and pillar are used for sample loading, and another stream and pillar are balanced wash-out simultaneously, with what is automatically switched
Function, and change pillar during convenient continuous production.
Comprise the following steps that：
(1) Fig. 1 connecting pipelines are pressed on chromatograph Explorer, second seven port valve is set to the Flow of system
Direction valves；
(2) system valve position is set to LOAD, during Up Flow, the feed liquid in sample pump enters post 3 and leads to from first seven
4. number mouth outflow of valve；The chromatographic steps such as the solution of systems pumps is balanced into post 4, drip washing, wash-out simultaneously flow through the inspection of system
Device and outlet valve are surveyed, sample pump and systems pumps can be operated simultaneously.
(3) system valve position is set to INJECT, during Down Flow, the feed liquid in sample pump enters post 4 and from second
4. number mouth outflow of seven port valves；The chromatographic steps such as the solution of systems pumps is balanced into post 3, drip washing, wash-out simultaneously flow through system
Detector and outlet valve, sample pump and systems pumps can be operated simultaneously
The state that (4) two seven port valves constantly repeat (2) and (3) can realize that two streams run two posts simultaneously parallel
Son, and with the function of automatically switching.
Embodiment 2
As shown in Fig. 3, Fig. 9 d, Fig. 9 e and Fig. 9 f, a kind of chromatograph reequips the method for perfusion culture purified, wherein institute
The method stated is：Replace two valves using seven port valves 1 and three eight ways valves 6,7,8 on chromatograph, by seven port valves 1 3.
Number mouth is connected the upper end for being connected to first pillar 3 with 5. number mouth threeway, and first lower end of pillar connects first eight and lead to
Valve 6, second upper end of pillar 4 is connected to by the 1. number mouth of seven port valves 1, second lower termination of pillar 4, second eight ways valve 7,
4. number mouth of seven port valves 1 is connected the upper end for being connected to the 3rd pillar 5, the 3rd lower end of pillar 5 with 6. number mouth threeway
The 3rd eight ways valve 8 is connect, a 2. number outlet for described three eight ways valves 6,7,8 is waste liquid port, three 1. number outlets of eight ways valve
One stream is converged into by threeway or four-way and is products export, is connected on the detector of system instrument that first eight ways valve 6 is 3.
Number mouth and 4. number mouth are connected to the 3rd pillar 5 and second upper end of pillar 4 by threeway respectively, second eight ways valve 7 3. number
Mouth and 4. number mouth are connected to the 3rd pillar 5 and first upper end of pillar 3, the 3rd 3. number mouth of eight ways valve 8 by threeway respectively
4. number mouth is connected to second pillar 4 and first upper end of pillar 3 by threeway respectively, systems pumps solution by seven port valves 1 7.
Number mouth enters, and sample pump solution is entered by the 2. number mouth of seven port valves 1, by switching seven port valves 1 and three valves of eight ways valve 6,7,8
Position state realizes that there are two streams to run three instruments of pillar simultaneously parallel, and a stream and pillar are used for sample loading,
Another stream and pillar are balanced wash-out simultaneously, and the 3rd pillar can receive the efflux from other two pillars,
And the circulation that can automatically switch of three pillars is carried out, so as to reach significantly cost-effective purpose under conditions of yield is not sacrificed.
As shown in figure 8, a kind of chromatograph reequips the method for perfusion culture purified, wherein described method is：In layer
Six seven port valves 1,2,15,16,17,18 are used in analyzer, wherein the 2. number mouth of three seven port valves 1,2,15 is connected to three respectively
The upper end of pillar 3,4,5, the 7. number mouth of the port valve 16,17,18 of the other three seven is connected to three lower ends of pillar 3,4,5 respectively, the
4. number mouth of one seven port valve, 1, second seven port valve 2 and the 3rd seven port valve 15 and the 4th seven port valve 16, the 5th seven lead to
6. number mouth threeway or the four-way of valve 17 and six seven port valves 18 connect together, and systems pumps solution is by first seven port valve 1, second
1. number mouth of individual seven port valve 2 and the 3rd seven port valves 15 enters, and sample pump solution is by first seven port valve, 1, second seven port valve 2
With the 3rd 3. number mouth entrance, described first seven port valve, 1, second seven port valve 2 and the 3rd seven port valves of seven port valves 15
15 5., 6., 7. a number mouth is shut, described the 4th seven port valves 16, the 5th seven port valves 17 and six seven port valves 18 5. number
Mouthful for waste liquid port, 1. number mouth is connected on the detector of system for products export, and 2. a number mouth is shut, the port valve of three after pillar seven
16th, 17,18 3. number mouth and 4. number mouth short circuit, by switching six valve position states of seven port valves 1,2,15,16,17,18, realize
Three instruments of pillar are run simultaneously parallel with two streams, a stream and pillar are used for sample loading, another stream
Wash-out is balanced simultaneously with pillar, the 3rd pillar can receive the efflux from other two pillars, and three pillars
The circulation that can automatically switch is carried out, so as to reach significantly cost-effective purpose under conditions of yield is not sacrificed.
Comprise the following steps that：
(1) Fig. 3 connecting pipelines are pressed on chromatograph Explorer.
(2) system valve position transfer sequence and state are as shown in table 1.
(3) the A-F states in the continuous repeat step (2) in system valve position can realize that two streams run three posts simultaneously parallel
Son, and with the function of automatically switching.Realize that every pillar is fully loaded significantly to be saved so as to be reached under conditions of yield is not sacrificed
The purpose of cost.
The system valve of table 1 position three refiting schemes of pillar of state and the port valve of function-seven and three eight ways valves
| Valve | A | B | C | D | E | F |
| Seven port valves | Load | Load | Waste | Waste | Inject | Inject |
| After first pillar-eight ways valve | 2 | 3 | 4 | 1 | 2 | 2 |
| After second pillar-eight ways valve | 3 | 1 | 2 | 2 | 2 | 4 |
| After 3rd pillar-eight ways valve | 2 | 2 | 2 | 3 | 4 | 1 |
| First pillar state | Loading | Loading | Drip washing after loading | Wash-out and releveling | From the leacheate of post 3 | Liquid is penetrated from post 1 |
| Second pillar state | Drip washing after loading | Wash-out and releveling | From the leacheate of post 1 | Liquid is penetrated from post 1 | Loading | Loading |
| 3rd pillar state | From the leacheate of post 2 | Liquid is penetrated from post 1 | Loading | Loading | Drip washing after loading | Wash-out and releveling |
Embodiment 3
As shown in figure 4, a kind of chromatograph reequips the method for perfusion culture purified, wherein described method is：In layer
Replace two valves, post upper end two eight ways valves 9,10, the 4th eight ways valve 9 using five eight ways valves 6,7,8,9,10 in analyzer
1., 2., 3. number mouth respectively with the 5th eight ways valve 10 3., 1., 2. a number mouth is connected and is connected to first by threeway
3, second pillar 4 of root pillar and the 3rd upper end of pillar 5, and first pillar, 3, second pillar 4 and the 3rd pillar 5
Lower end connects first eight ways valve, 6, second eight ways valve 7 and the 3rd eight ways valve 8 respectively, first 3. number mouth of eight ways valve 6 and
4. number mouth is connected to the 3rd pillar 5 and second upper end of pillar 4 by threeway respectively, the 3. number mouth of second eight ways valve 7 and 4.
Number mouth is connected to the 3rd pillar 5 and first upper end of pillar 3 by threeway respectively, the 3. number mouth of the 3rd eight ways valve 8 and 4. number
Mouth is connected to second pillar 4 and first upper end of pillar 3 by threeway respectively, first, second and the 3rd eight ways valve 6,
7th, a 8 2. number outlet is waste liquid port, and 1. number mouth converges into a stream and is products export, is connected to the inspection of instrument by threeway or four-way
Survey on device, systems pumps solution is entered by the 4th center of eight ways valve 9, and sample pump solution is entered by the 5th center of eight ways valve 10,
Realize that two streams are parallel by the valve position state for switching eight ways valve, three pillars run two-by-two, and a stream and pillar are used for
Sample loading, another stream and pillar are balanced wash-out simultaneously, with the function of automatically switching, and during convenient continuous production
Pillar is changed, or three instruments of pillar is run simultaneously parallel with two streams, a stream and pillar are used for sample loading,
Another stream and pillar are balanced wash-out simultaneously, and the 3rd pillar can receive the efflux from other two pillars,
And the circulation that can automatically switch of three pillars is carried out, so as to reach significantly cost-effective purpose under conditions of yield is not sacrificed.
Comprise the following steps that：
(1) Fig. 4 connecting pipelines are pressed on chromatograph Explorer.
(2) system valve position transfer sequence and state are as shown in table 2.
(3) the A-F states in the continuous repeat step (2) in system valve position can realize that two streams run three posts simultaneously parallel
Son, and with the function of automatically switching.Realize that every pillar is fully loaded significantly to be saved so as to be reached under conditions of yield is not sacrificed
The purpose of cost.
The system valve of table 2 position state and logical (or equivalent) three refiting schemes of pillar of function-five eight
| Valve | A | B | C | D | E | F |
| Three eight ways valves of post front end-two | 1 | 1 | 3 | 3 | 2 | 2 |
| After first pillar-eight ways valve | 2 | 3 | 4 | 1 | 2 | 2 |
| After second pillar-eight ways valve | 3 | 1 | 2 | 2 | 2 | 4 |
| After 3rd pillar-eight ways valve | 2 | 2 | 2 | 3 | 4 | 1 |
| First pillar state | Loading | Loading | Drip washing after loading | Wash-out and releveling | From the leacheate of post 3 | Liquid is penetrated from post 1 |
| Second pillar state | Drip washing after loading | Wash-out and releveling | From the leacheate of post 1 | Liquid is penetrated from post 1 | Loading | Loading |
| 3rd pillar state | From the leacheate of post 2 | Liquid is penetrated from post 1 | Loading | Loading | Drip washing after loading | Wash-out and releveling |
Embodiment 4
As shown in figure 5, a kind of chromatograph reequips the method for perfusion culture purified, wherein described method is：New
Two valves are used on type chromatograph, when two described valves are seven port valve 11,12, first pillar 3 and second pillar 4 are distinguished
It is connected between two LoopE mouthfuls and LoopF mouthfuls of seven port valves 11,12, first Syp mouthfuls of seven port valves 11 and second seven port valve
Col mouthfuls connection, first W2 mouthfuls of seven port valves 11 is connected with second the Sap of seven port valves mouthfuls, and systems pumps solution is by first
Col mouthfuls of entrance of seven port valves 11, sample pump solution is by first Sap mouthfuls of entrance of seven port valves 11, two W1 of seven port valves 11,12
Be waste liquid port, second Syp mouthfuls of seven port valves 12 be products export, be connected on the detector of system instrument, by switching two
The valve position state of individual seven port valve 11,12, realizes that two streams run two pillars simultaneously parallel, and a stream and pillar are used for sample
Product loading, another stream and pillar are balanced wash-out simultaneously, and with the function of automatically switching.
As shown in Fig. 6, Fig. 9 a and Fig. 9 b, a kind of chromatograph reequips the method for perfusion culture purified, wherein described
Method is：Use two valves on new chromatograph, described two valves one of them be seven port valves 11 and another be post position
Valve 13, LoopE mouthfuls of seven described port valves 11 connects first upper end of pillar 3, and W1 mouthfuls and W2 with LoopF mouthfuls of short circuit, Col mouthfuls
Mouth is connected to second upper end of pillar 4 by threeway, and 2A mouthfuls and 1B mouthfuls of described post position valve 13 is connected to first by threeway
The lower end of root pillar 3, second lower end of pillar 4 is connected to by the 2B and 1A of post position valve by threeway, and systems pumps solution is led to by seven
Syp mouthfuls of entrance of valve 11, sample pump solution is entered by Sap mouthfuls of seven port valves 11, and the valve import of post position is waste liquid port, post position valve outlet
For products export, it is connected on the detector of system instrument, by switching the valve position state of seven port valves 11 and the difference of post position valve 13
Post position, realizes that two streams run two pillars simultaneously parallel, and a stream and pillar are used for sample loading, another stream and
Pillar is balanced wash-out simultaneously, and with the function of automatically switching.
Comprise the following steps that：
(1) Fig. 6 connecting pipelines are pressed on new chromatograph Pure M.
(2) system valve position is set to Direct Inject, during a post position, the feed liquid in sample pump enter post 3 and from
The import outflow of post position valve；The chromatographic steps such as the solution of systems pumps is balanced into post 4, drip washing, wash-out simultaneously flow through system
Detector and outlet valve, sample pump and systems pumps can be operated simultaneously.
(3) system valve position is set to Inject, during No. two post positions, the feed liquid in sample pump enters post 4 and from post position valve
Import outflow；The chromatographic steps such as the solution of systems pumps is balanced into post 3, drip washing, wash-out simultaneously flow through the detector of system
And outlet valve, sample pump and systems pumps can be operated simultaneously
(4) the continuous state for repeating (2) and (3) in system valve position can realize that two streams run two pillars simultaneously parallel,
And with the function of automatically switching.
Embodiment 5
As shown in Fig. 7, Fig. 9 a, Fig. 9 b and Fig. 9 c, described two valves one of them be post position valve 14 and another be seven
Port valve 11,3A mouthfuls of described post position valve 14 with 1B mouthfuls by threeway be connected to first upper end of pillar 3,1A mouthfuls pass through with 2B mouthfuls
Threeway is connected to the upper end of second pillar 4 and 2A mouthfuls is connected to the 3rd upper end of pillar 5 with 3B mouthful by threeway, and described seven are logical
LoopE mouthfuls of valve 11 with LoopF mouthful by threeway be connected to first lower end of pillar 3, W1 mouthfuls and W2 mouthfuls be connected to second by threeway
The lower end of root pillar 4 and Col mouthfuls are connected to first lower end of pillar 3, and systems pumps solution is entered by the import of post position valve, sample pump solution
Entered by the outlet of post position valve, Sap mouthfuls of seven described port valves 11 is waste liquid port, Syp mouthfuls of seven described port valves 11 is product
Export, be connected on the detector of system instrument, it is real by switching the valve position state of seven port valves 11 and the different posts position of post position valve 14
Existing two streams are parallel, and three pillars run two-by-two, and a stream and pillar are used for sample loading, and another stream and pillar are same
When be balanced wash-out, with the function of automatically switching, and pillar is changed during convenient continuous production.
Two described valves may also be two post position valves or two seven port valves, and its connection stream leads to post position valve 14 and seven
The stream of valve 11 is consistent, by switching the valve position state of two seven port valves or two post position valves, realizes that two streams are parallel, three posts
Son runs two-by-two, and a stream and pillar are used for sample loading, and another stream and pillar are balanced wash-out simultaneously, with certainly
The function of dynamic switching, and change pillar during convenient continuous production.
Comprise the following steps that：
(1) Fig. 7 connecting pipelines are pressed on new chromatograph Pure M instruments.
(2) system valve position is set to post position, during Inject, the feed liquid in sample pump enters post 4 and from seven port valves
Sap mouthfuls outflow；The chromatographic steps such as the solution of systems pumps is balanced into post 3, drip washing, wash-out simultaneously flow through the detector of system
And outlet valve, sample pump and systems pumps can be operated simultaneously.
(3) system valve position is set to No. two posts position, during Direct Inject, the feed liquid in sample pump enter post 5 and from
Syp mouthfuls of outflow of seven port valves；The chromatographic steps such as the solution of systems pumps is balanced into post 4, drip washing, wash-out simultaneously flow through system
Detector and outlet valve, sample pump and systems pumps can be operated simultaneously.
(4) system valve position is set to No. three post positions, during Sample Pump Load, the feed liquid in sample pump enters post 3
And flowed out from Sap mouthfuls of seven port valves；The chromatographic steps such as the solution of systems pumps is balanced into post 5, drip washing, wash-out simultaneously flow through and are
The detector and outlet valve of system, sample pump and systems pumps can be operated simultaneously.
(5) the continuous repetition (2) in system valve position, (3), the state of (4) can realize that two streams run three posts simultaneously parallel
Son, and with the function of automatically switching.
Claims (9)
1. a kind of chromatograph reequips the method for perfusion culture purified, it is characterised in that：Described method is：On chromatograph
Using two valves, when two described valves are seven port valves (1,2), by the 5. number mouth of first seven port valve (1) and 7. number mouth is distinguished
With second 3. number mouth of seven port valves (2), 1. a number mouth is connected, and a 5. number mouth for second described seven port valve (2) is shut, described
Second 7. number mouth of seven port valves (2) is products export, is connected on the detector of system instrument, described two seven port valves (1,
2) a 4. number mouth is waste liquid port, and two pillars (3,4) are attached separately to 2. and 6. between number mouth of two seven port valves, and systems pumps are molten
Liquid is entered by the 1. number mouth of first seven port valve (1), and sample pump solution is entered by the 3. number mouth of first valve (1), by switching
The valve position state of two seven port valves (1,2) realizes that two streams run two pillars simultaneously parallel, and a stream and pillar are used for
Sample loading, another stream and pillar are balanced wash-out simultaneously, and with the function of automatically switching.
2. a kind of chromatograph as claimed in claim 1 reequips the method for perfusion culture purified, it is characterised in that：Described
When two valves are seven port valves (1,2), first 3. number mouth of seven port valves (1) are connected with a 5. number mouth threeway and are connected to first
The upper end of pillar (3), second 3. number mouth of seven port valves (2) is connected with a 5. number mouth threeway and is connected to first pillar (3)
Lower end, the 1. number mouth of first seven port valve (1) is connected to second upper end of pillar (4), second 1. number mouth of seven port valves
Second lower end of pillar (4) is connected to, first 4. number mouth of seven port valves is connected with a 6. number mouth threeway and is connected to the 3rd
The upper end of pillar (5), second 4. number mouth of seven port valves is connected with a 6. number mouth threeway and is connected under the 3rd pillar (5)
End, second 2. number mouth of seven port valves (2) is waste liquid port, and second 7. number mouth of seven port valves (2) is products export, is connected to system
On the detector of instrument, systems pumps solution is entered by the 7. number mouth of first seven port valve (1), and sample pump solution is led to by first seven
2. number mouth of valve (1) enters, and realizes that two streams are parallel by the valve position state for switching two seven port valves (1,2), three pillars
Run two-by-two, a stream and pillar are used for sample loading, and another stream and pillar are balanced wash-out simultaneously, with automatic
The function of switching, and change pillar during convenient continuous production.
3. a kind of chromatograph as claimed in claim 1 reequips the method for perfusion culture purified, it is characterised in that：Described
Two valves are replaced with seven port valve (1) and three eight ways valves (6,7,8), and the 3. number mouth of seven port valves (1) is used into three with a 5. number mouth
Lead to the upper end for connecing and being connected to first pillar (3), first lower end of pillar connects first eight ways valve (6), by seven port valves
(1) 1. number mouth is connected to second upper end of pillar (4), second lower termination of pillar (4), second eight ways valve (7), by seven
4. number mouth of port valve (1) is connected the upper end for being connected to the 3rd pillar (5) with 6. number mouth threeway, under the 3rd pillar (5)
The 3rd eight ways valve (8) is terminated, a 2. number outlet for described three eight ways valves (6,7,8) is waste liquid port, and three eight ways valves are 1.
Number outlet by threeway or four-way converge into a stream for products export, be connected on the detector of system instrument, first eight is logical
3. number mouth of valve (6) and 4. number mouth are connected to the 3rd pillar (5) and second pillar (4) upper end, second by threeway respectively
3. number mouth of eight ways valve (7) and 4. number mouth are connected to the 3rd pillar (5) and first pillar (3) upper end by threeway respectively, the
3. number mouth of three eight ways valves (8) and 4. number mouth is connected on second pillar (4) and first pillar (3) by threeway respectively
End, systems pumps solution is entered by the 7. number mouth of seven port valves (1), and sample pump solution is entered by the 2. number mouth of seven port valves (1), by cutting
The valve position state of seven port valves (1) and eight ways valve (6,7,8) is changed, realizes that there are two streams to run three instrument of pillar simultaneously parallel
Device, a stream and pillar are used for sample loading, and another stream and pillar are balanced wash-out simultaneously, and the 3rd pillar receives
Efflux from other two pillars, and the circulation that can automatically switch of three pillars carried out, and yield is not being sacrificed so as to reach
Under the conditions of significantly cost-effective purpose.
4. a kind of chromatograph as claimed in claim 1 reequips the method for perfusion culture purified, it is characterised in that：Described
Two valves are replaced with five eight ways valves (6,7,8,9,10), the eight ways valve of post upper end two (9,10), the 4th eight ways valve (9)
1., 2., 3. number mouth respectively with the 5th eight ways valve (10) 3., 1., 2. a number mouth is connected and is connected to first by threeway
Root pillar (3), second pillar (4) and the 3rd pillar (5) upper end, and first pillar (3), second pillar (4) and
The lower end of three pillars (5) connects first eight ways valve (6), second eight ways valve (7) and the 3rd eight ways valve (8), first respectively
3. number mouth of individual eight ways valve (6) and 4. number mouth are connected to the 3rd pillar (5) and second pillar (4) upper end by threeway respectively,
3. number mouth of second eight ways valve (7) and 4. number mouth is connected on the 3rd pillar (5) and first pillar (3) by threeway respectively
End, the 3. number mouth of the 3rd eight ways valve (8) and 4. number mouth is connected to second pillar (4) and first pillar by threeway respectively
(3) upper end, a 2. number outlet for first, second and the 3rd eight ways valve (6,7,8) is waste liquid port, 1. number mouth by threeway or
Four-way converges into a stream and is products export, is connected on the detector of instrument, and systems pumps solution is by the 4th eight ways valve (9) center
Into sample pump solution is entered by the 5th eight ways valve (10) center, by switching five valves of eight ways valve (6,7,8,9,10)
Position state realizes that two streams are parallel, and three pillars run two-by-two, and a stream and pillar are used for sample loading, another stream
Wash-out is balanced simultaneously with pillar, with the function of automatically switching, and pillar is changed during convenient continuous production, or with two
Stream simultaneously runs three instruments of pillar simultaneously, and a stream and pillar are used for sample loading, and another stream and pillar are simultaneously
It is balanced wash-out, the 3rd pillar receives the efflux from other two pillars, and three pillars can automatically switch circulation
Carry out, so as to reach the function of significantly cost-effective purpose under conditions of yield is not sacrificed.
5. a kind of chromatograph as claimed in claim 1 reequips the method for perfusion culture purified, it is characterised in that：Described
Two valves are replaced with six seven port valves (1,2,15,16,17,18), wherein a 2. number mouth for three seven port valves (1,2,15) connects respectively
In the upper end of three pillars (3,4,5), the 7. number mouth of the port valve of the other three seven (16,17,18) be connected to respectively three pillars (3,4,
5) lower end, the 4. number mouth of first seven port valve (1), second seven port valve (2) and the 3rd seven port valve (15) and the 4th seven
6. number mouth threeway or the four-way of port valve (16), the 5th seven port valve (17) and six seven port valves (18) connect together, systems pumps
Solution is entered by the 1. number mouth of first seven port valve (1), second seven port valve (2) and the 3rd seven port valve (15), and sample pump is molten
Liquid is entered by the 3. number mouth of first seven port valve (1), second seven port valve (2) and the 3rd seven port valve (15), and described first
Individual seven port valve (1), second seven port valve (2) and the 3rd seven port valve (15) 5., 6., 7. a number mouth is shut, described the 4th
Seven port valves (16), the 5th seven port valve (17) and six 5. number mouths of seven port valves (18) be waste liquid port, 1. number mouth be products export
It is connected on the detector of system, 2. a number mouth is shut, the 3. number mouth of the port valve of three after each pillar seven and 4. number mouth short circuit pass through
The valve position state of six seven port valves (1,2,15,16,17,18) of switching, realizes that there are two streams to run three posts simultaneously parallel
The instrument of son, a stream and pillar are used for sample loading, and another stream and pillar are balanced wash-out, the 3rd post simultaneously
Son receives the efflux from other two pillars, and the circulation that can automatically switch of three pillars is carried out, and is not being sacrificed so as to reach
Significantly cost-effective purpose under conditions of yield.
6. a kind of chromatograph reequips the method for perfusion culture purified, it is characterised in that：Described method is：On chromatograph
Using two valves, when two described valves are seven port valves (11,12), first pillar (3) and second pillar (4) are connected to respectively
Between two LoopE mouthfuls and LoopF mouthfuls of seven port valves (11,12), first Syp mouthfuls of seven port valves (11) and second seven port valve
Col mouthfuls connection, first W2 mouthfuls of seven port valves (11) is connected with second the Sap of seven port valves mouthfuls, and systems pumps solution is by first
Col mouthfuls of entrance of individual seven port valve (11), sample pump solution is by first Sap mouthfuls of entrance of seven port valves (11), two seven port valves
W1 mouthfuls of (11,12) is waste liquid port, second Syp mouthfuls of seven port valves (12) be products export, be connected to the detection of system instrument
On device, by switching two valve position states of seven port valves (11,12), realize that two streams run two pillars, one simultaneously parallel
Bar stream and pillar are used for sample loading, and another stream and pillar are balanced wash-out simultaneously, and with the work(for automatically switching
Energy.
7. a kind of chromatograph as claimed in claim 6 reequips the method for perfusion culture purified, it is characterised in that：Described
One of them is seven port valves (11) for two valves, when another is post position valve (13), LoopE mouthful of described seven port valves (11) with
LoopF mouthfuls of short circuit, first pillar (3) upper end of Col mouthfuls of connection, and W1 mouthful be connected to second pillar with W2 mouthfuls by threeway
(4) upper end, 2A mouthfuls and 1B mouthfuls of described post position valve (13) is connected to first pillar (3) lower end by threeway, by post position valve
2B and 1A second lower end of pillar (4) is connected to by threeway, systems pumps solution by seven port valves (11) Syp mouthfuls of entrance,
Sample pump solution is entered by Sap mouthfuls of seven port valves (11), and the valve import of post position is waste liquid port, and post position valve outlet is products export, connects
In on the detector of system instrument, by switching the valve position state of seven port valves (11) and the different posts position of post position valve (13), realize
Two streams run two pillars simultaneously parallel, and a stream and pillar are used for sample loading, and another stream and pillar are simultaneously
Wash-out is balanced, and with the function of automatically switching.
8. a kind of chromatograph as claimed in claim 6 reequips the method for perfusion culture purified, it is characterised in that：Described
Two valves one of them be post position valve (14) and another when being seven port valves (11), 3A mouthfuls and 1B mouthfuls of described post position valve (14)
By threeway be connected to first pillar (3) upper end, 1A mouthfuls second upper end of pillar (4) is connected to by threeway with 2B mouthfuls,
2A mouthfuls is connected to the 3rd pillar (5) upper end with 3B mouthfuls by threeway, and LoopE mouthfuls of described seven port valves (11) leads to LoopF mouthfuls
Cross threeway be connected to first pillar (3) lower end, W1 mouthfuls second pillar (4) lower end is connected to by threeway with W2 mouthfuls and Col mouthfuls connects
In the 3rd post (5) lower end, systems pumps solution is entered by the import of post position valve, and sample pump solution is entered by the outlet of post position valve,
Sap mouthfuls of described seven port valves (11) is waste liquid port, Syp mouthful of described seven port valves (11) be products export, be connected to system instrument
On the detector of device, by switching the valve position state of seven port valves (11) and the different posts position of post position valve (14), two streams are realized
Parallel, three pillars run two-by-two, and a stream and pillar are used for sample loading, and another stream and pillar are balanced simultaneously
Wash-out, with the function of automatically switching, and changes pillar during convenient continuous production.
9. a kind of chromatograph according to claim 8 reequips the method for perfusion culture purified, it is characterised in that：It is described
Two valves for two posts position valves or two seven port valves when, its connection stream and a post valve (14) and seven port valve (11)
Stream is consistent, by switching the different posts position of two valve of seven port valves states or two post position valves, realizes that two streams are parallel,
Three pillars run two-by-two, and a stream and pillar are used for sample loading, and another stream and pillar are balanced wash-out simultaneously,
With the function of automatically switching, and pillar is changed during convenient continuous production.
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| CN107485891B (en) * | 2017-07-20 | 2020-04-21 | 上海药明生物技术有限公司 | Improved chromatographic device and method for continuous flow chromatography |
| CN111068360A (en) * | 2018-10-22 | 2020-04-28 | 信达生物制药(苏州)有限公司 | Combined chromatographic device and continuous flow chromatographic method |
| WO2020258705A1 (en) * | 2019-06-28 | 2020-12-30 | 信达生物制药(苏州)有限公司 | Modular chromatography device |
| CN114072217A (en) * | 2019-06-28 | 2022-02-18 | 信达生物制药(苏州)有限公司 | Combined chromatography device, seamless connection chromatography method and biological agent purification method |
| CN113332753B (en) * | 2021-05-14 | 2022-11-01 | 浙江金仪盛世生物工程有限公司 | Continuous flow chromatography system and chromatography method |
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