CN2377250Y - External pressure type hollow fibre column - Google Patents
External pressure type hollow fibre column Download PDFInfo
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- CN2377250Y CN2377250Y CN 99202751 CN99202751U CN2377250Y CN 2377250 Y CN2377250 Y CN 2377250Y CN 99202751 CN99202751 CN 99202751 CN 99202751 U CN99202751 U CN 99202751U CN 2377250 Y CN2377250 Y CN 2377250Y
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- hollow fiber
- room
- external
- compression type
- tubing string
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Abstract
The utility model relates to an external pressure type hollow fiber column, which comprises a pipe column provided with a solution gateway and a waste liquid/washing liquid gateway. An accommodation room is divided by two epoxy resin isolation layers positioned at both ends of the pipe column accommodation room into a top waste liquid/washing liquid room and a bottom waste liquid/washing liquid room, and a middle renaturation protein liquid room. A plurality of hollow fiber film pipes are arranged between the two epoxy resin isolation layers to communicate the top and the bottom waste liquid/washing liquid rooms. The hollow fiber film pipe is composed of a support layer arranged on the inner side of the film pipe and a function select layer arranged on the outer side of the film pipe. The epoxy resin isolation layer is funnel-shaped. The funnel walls are respectively inclined towards the interior of the accommodation room to form an obtuse angle of more than 90 DEG with the pipe column wall.
Description
The utility model relates to biological technical field, specifically belongs to a kind of external-compression type hollow fiber column that is used for the denatured protein renaturation.
The protein of multiple economically valuable is very low at occurring in nature or content, and perhaps extraction cost is very high, thus at present using gene engineering technique to obtain proteinic progress very fast.Wherein especially with the escherichia expression system research object of attaching most importance to, because intestinal bacteria genetic background is comparatively clear, and as the plasmid of expression vector than horn of plenty, bacterium is growth easily again, therefore, the intestinal bacteria system is an effective system of expressing exogenous protein.But be that the exogenous protein overwhelming majority that host's system is expressed exists with the inclusion body form with intestinal bacteria, though target protein matter is purer in the inclusion body, generally exist in insoluble non-activity mode this moment.In order to obtain activated target protein matter, dissolve inclusion body with denaturing agent, remove denaturing agent then under certain condition, obtain activated protein, also be the renaturation of denatured protein.In the past, the renaturation of denatured protein was to carry out in dialysis tubing, or the large volume dilution is to obtain the bioactive protein of tool, and this mode is operated loaded down with trivial details, and the time is long, and the active result yield is low, and influences the later separation purifying, thereby can't scale operation.The ultrafiltration mode of Chu Xianing can be fit to from the laboratory to producing different needs at all levels, wherein especially to adopt hollow fiber column more general in recent years.But as Figure 1-3, the hollow fiber column of prior art is inner pressed, it is the inboard that the selection function layer 102 of ultra-filtration membrane in the hollow fiber column is located at hollow fiber film tube 10, because the internal diameter of thread hollow fiber film tube 10 is generally below 1mm, because of the film pipe too thin, in this inner pressed hollow fiber column, along with recombinant protein solution flows in film pipe 10, the reduction of denaturing agent concentration is very fast, then correctly the intermediate of renaturation is assembled in hollow fiber film tube 10 easily, and then produces precipitation, thereby stops up hollow fiber film tube, cause the ultrafiltration decrease in efficiency, and the damage of hollow fiber column.And the Resins, epoxy sealing coat 6 of existing hollow fiber column becomes 90 ° of angles with the hollow fiber column perisporium, thereby has formed the dead angle on top up and down, is unfavorable for the flowing of solution in the room, turnover.
The purpose of this utility model is to overcome the deficiencies in the prior art, provide a kind of in the protein renaturation process, can reduce recombinant protein solution and in hollow fiber film tube, flow the time correctly the various intermediates of renaturation produce and precipitate, and can make the more abundant external-compression type completely of protein renaturation hollow fiber column.
The purpose of this utility model is achieved through the following technical solutions:
The external-compression type hollow fiber column, it mainly comprises: a tubing string, this tubing string contains cylindrical cell, respectively is provided with a solution gangway on it on the lower wall central position, is provided with some hollow fiber film tubes and diepoxy resin sealing coat in the room, it is characterized in that:
Described Resins, epoxy sealing coat is installed in the two ends of room, room is divided into waste liquid/cleaning liquid chamber and middle recombinant protein liquid chamber up and down, at lower wall on the tubing string with respectively be provided with one waste liquid/scavenging solution gangway on the tubing string perisporium between the Resins, epoxy sealing coat up and down, described hollow fiber film tube is installed between the diepoxy resin sealing coat, and is communicated with waste liquid/cleaning liquid chamber up and down;
Described hollow fiber film tube selects layer to form by a supporting layer and a function, and wherein, supporting layer is in the inboard of film pipe, and function is selected the outside of layer at the film pipe.
Above-mentioned external-compression type hollow fiber column, wherein: described up and down Resins, epoxy sealing coat is funnel-form, and its headwall tilts in room respectively, makes itself and the angle of inclination beta of tubing string internal perisporium formation one greater than 90 °;
Above-mentioned external-compression type hollow fiber column, wherein: described angle of inclination beta is good with 110-135 °.
Above-mentioned external-compression type hollow fiber column, wherein: described hollow fiber film tube to be installing with the mode of tubing string central axes, and is crossed as the obtuse angle with the Resins, epoxy sealing coat.
Above-mentioned external-compression type hollow fiber column, wherein: described hollow-fibre membrane bore is the 0.3-0.5 millimeter, its molecular retention value is MW 1000-50,000 dalton (Dalton).
Above-mentioned external-compression type hollow fiber column, wherein: the material of described hollow fiber film tube is a polysulfone material.
The utility model is because the inboard of taking the supporting layer in the hollow fiber film tube is located at the film pipe, the function selection is placed on the design in the outside of film pipe, like this, the saturating filter of sex change factor in the recombinant protein solution is flowed in the film pipe, in addition owing to flow in the recombinant protein liquid chamber of recombinant protein solution outside the film pipe, because film ligament space is than much bigger in the film pipe, therefore can not form the concentration gradient of materials such as tangible denaturing agent in the recombinant protein solution, help proteinic abundant renaturation, can significantly reduce the precipitation that in renaturation process, forms; Simultaneously, recombinant protein solution flows outside the tunica fibrosa pipe, can reduce working pressure, reduces the absorption of protein on the film pipe; In addition, the emitting fully of protein solution after also helping renaturation manipulation and finishing, and the protein precipitation on the film pipe can fully be washed out when cleaning, and improves proteic renaturation yield and makes hollow fiber column regenerated effect preferably thereby reach.
Simultaneously, because the sealing coat of Resins, epoxy up and down of the utility model room tilts in room respectively, make itself and tubing string internal perisporium form one greater than 90 ° obtuse angle, thereby the dead angle that can reduce room, the emitting of protein solution after being convenient to renaturation manipulation and finishing, and fully the washing out of protein precipitation when cleaning.
The utility model is described in further detail below in conjunction with embodiment that accompanying drawing and accompanying drawing provide:
Fig. 1 is the structural representation of prior art inner pressed hollow fiber column;
Fig. 2 is the A-A sectional view of Fig. 1;
Fig. 3 is the local enlarged diagram of hollow fiber film tube among Fig. 1;
Fig. 4 is the structural representation of the utility model external-compression type hollow fiber column;
Fig. 5 is the A-A sectional view of Fig. 4;
Fig. 6 is the local enlarged diagram of hollow fiber film tube among Fig. 4.
Fig. 4-6 is the structural representation of the utility model external-compression type hollow fiber column.
Be illustrated in figure 4 as the external-compression type hollow fiber column that is used for metaprotein solution renaturation that a pmma material is made, comprise a tubing string 100, it comprises a cylindrical cell 1, respectively offers a recombinant protein solution gangway 2,3 in the corresponding position, the centre of end wall up and down of post;
Upper and lower ends respectively is provided with a Resins, epoxy sealing coat 6,6 ' in the cylindrical cell 1, with room be divided into up and down waste liquid/cleanings liquid chamber 7,7 ' with intermediary recombinant protein liquid chamber 8.On waste liquid up and down/cleaning liquid chamber 7,7 ' tubing string perisporium, respectively be provided with one waste liquid/scavenging solution gangway 4,5.Described 2-5 is equipped with screw thread on each gangway, and the various tubings that can be spirally connected are equipped with corresponding adjustable clamp (not shown) on each tubing.
Resins, epoxy sealing coat 6,6 ' is funnel-form up and down, its headwall tilts in room respectively, and make Resins, epoxy sealing coat 6,6 ' and the angle that forms of post jamb be obtuse angles beta greater than 90 °, (being 120 ° angle shown in the figure) thus can reduce because 90 ° of angles form the difficult cleaning positions that cause at dead angles, or renaturation manipulation finish after the emitting fully of recombinant protein solution.Like this, recombinant protein solution solution in room is unobstructed when being more conducive to renaturation manipulation flows, renaturation manipulation is finished after the cleaning of room more abundant.
In recombinant protein liquid chamber 8, between diepoxy resin sealing coat 6,6 ', utmost point multi-filament shape hollow fiber film tube 10 is installed, they are to install with the mode of tubing string central axes, its two ends penetrate respectively and are fixed in up and down on the Resins, epoxy sealing coat 6,6 ', and up and down waste liquid/cleanings liquid chamber 7,7 ' connection, these hollow fiber film tubes 10 also are crossed as the obtuse angle with Resins, epoxy sealing coat 6,6 '.Shown in Fig. 4-5,9 is the space between the hollow fiber film tube 10.
As shown in Figure 6, hollow fiber film tube 10 is a kind of superfine hollow fibre filaments of being made by polysulfone material, it selects layer 102 to form by a supporting layer 101 and a function, wherein, supporting layer 101 is in the inboard of hollow fiber film tube 10, function selects layer 102 in the outside of hollow fiber film tube 10, promptly towards a side of recombinant protein solution.The molecular interception value of this film pipe can be MW1,000-50, and 000Dalton specifically selects for use by proteinic molecular weight decision, and the membrane material tool hangs down the protein adsorption characteristic, and chemical stability is better.Because the supporting layer 101 of hollow fiber film tube 10 is located at its inboard, and select layer 102 to be located at the outside of hollow fiber film tube 10 function, can cause small-molecule substances such as the denaturing agent 9 saturating filters outside the film pipe in the recombinant protein solution to flow in the film pipe 10, protein molecule in the recombinant protein solution is owing to flow in the big then space 9 outside filamentary fibers film pipe of molecular weight, (be depicted as inner pressed hollow fiber column in the prior art as arrow among Fig. 3, the recombinant protein solution C flows in film pipe 10; Arrow is depicted as the utility model external-compression type hollow fiber column among Fig. 6, and recombinant protein solution is to flow in the gap outside the film pipe, and waste liquid B is two-way flow in film pipe 10 then.) like this, can make that removing by adjustable clamp of sex change factor obtains more suitable control in the recombinant protein solution, make the concentration gradient that can not form materials such as tangible denaturing agent in the recombinant protein solution, help proteinic abundant renaturation, can significantly reduce the precipitation that in renaturation process, forms; Simultaneously, recombinant protein solution flows outside the tunica fibrosa pipe, can reduce working pressure, reduces the absorption of protein on the film pipe; In addition, the emitting fully of protein solution after helping renaturation manipulation too and finishing, and fully being washed out of the protein precipitation on the film pipe when cleaning are improved proteic renaturation yield and are made hollow fiber column regenerated effect preferably thereby reach.
During renaturation manipulation, protein solution enters recombinant protein liquid chamber 8 from recombinant protein solution gangway 2 under the effect of gravity or pump, be full of chamber 8, regulate the degree of tightness of recombinant protein solution gangway 3 and waste liquid/scavenging solution gangway 5 and 4 places control folder, the speed in the hollow fiber film tube 10 is filtered in the medium and small molecule degeneration agent of may command recombinant protein solution thoroughly, thereby can the speed of different proteins, different renaturation stage removal denaturing agent be control effectively.Renaturation manipulation after finishing is tightened the clip at 3 places, recombinant protein solution gangway, recombinant protein solution is concentrated, thereby can make things convenient for subsequent disposal greatly.When the recombinant protein solution concentration behind certain volume, tighten the clip at waste liquid/5,4 places, scavenging solution gangway, loosen the clip at 3 places, recombinant protein solution gangway, emit from recombinant protein solution gangway 2, the damping fluid that is pressed into certain volume by recombinant protein solution gangway 3 washs recombinant protein liquid chamber 8 then, and washings is emitted and reclaimed from recombinant protein solution gangway 2.
To above-mentioned hollow fiber column 100 of having emitted recombinant protein solution, close waste liquid/scavenging solution gangway 4, be pressed into certain damping fluid from waste liquid/scavenging solution gangway 5, after sticking to hollow fiber film tube 10 outsides, being the albumen flushing on the function selection layer 102, emit recovery by recombinant protein solution gangway 2.
To the cleaning of hollow fiber column room 1, can close waste liquid/scavenging solution gangway 5, pump into certain density scavenging solution from waste liquid/scavenging solution gangway 4 and (, reach soda acid such as 1N NaOH, 1N HCl etc. as 6N guanidine hydrochloride solution, 8M urea soln, 2%SDS.) carry out that original position is cleaned or sterilization; Use buffer solution for cleaning afterwards again, after cleaning finishes,, close 2,3,5 gangways, be pressed into a certain amount of 20% spirituous solution by waste liquid/scavenging solution gangway 4 again, under 4-8 ℃ of condition, preserve by tightening the adjustable clamp at each relevant position.
The utility model external-compression type hollow fiber column except that the renaturation manipulation that can be applicable to inclusion body protein matter as mentioned above, also can be applicable to following several respects:
1. be used for useless proteic renaturation: useless albumen refers to protein renaturation, sex change not having of producing of inclusion body protein matter renaturation manipulation process, that exist with the aggregate and precipitate form.
2. at aspects such as concentrating of the conversion of proteinic chemically modified transformation, protein soln system and protein soln huge application potential is arranged also: proteinic chemically modified transformation is to obtain the proteinic a kind of strong means of new function, modify by PEG such as collagenase, IL-2, rhTPO etc., both can reduce its immunogenicity, can prolong it again in the intravital transformation period of people, thereby strengthen greatly by the use value of modifying protein and the scope of application.In proteinic chemically modified transformation process, the adding of modifier and remove and all should obtain certain control.
3. the application in enzyme reactor: the utility model is combined in the enzyme reaction device of water or organic phase, can obtains enzyme and reactant and product separate easily; Enzyme, reactant and product separate easily; Enzyme reaction such as can carry out continuously at effect.
Claims (7)
1. external-compression type hollow fiber column, it mainly comprises: a tubing string, this tubing string contains cylindrical cell, respectively is provided with a solution gangway on it on the lower wall central position, and device has some hollow fiber film tubes and diepoxy resin sealing coat in the room, it is characterized in that:
Described Resins, epoxy sealing coat is installed in the two ends of room, and room is divided into up and down waste liquid/cleaning liquid chamber and middle recombinant protein liquid chamber, at lower wall on the tubing string with respectively be provided with one waste liquid/scavenging solution gangway on the tubing string perisporium between the Resins, epoxy sealing coat up and down, described hollow fiber film tube is installed between the diepoxy resin sealing coat, and is communicated with waste liquid/cleaning liquid chamber up and down;
Described hollow fiber film tube selects layer to form by a supporting layer and a function, and wherein, supporting layer is in the inboard of film pipe, and function is selected the outside of layer at the film pipe.
2. external-compression type hollow fiber column according to claim 1 is characterized in that: described up and down Resins, epoxy sealing coat is funnel-form, and its headwall tilts in room respectively, makes itself and the angle of inclination beta of tubing string internal perisporium formation one greater than 90 °;
3. external-compression type hollow fiber column according to claim 1 is characterized in that: described angle of inclination beta is 110-135 °.
4. external-compression type hollow fiber column according to claim 1 is characterized in that: described hollow fiber film tube to be installing with the mode of tubing string central axes, and is crossed as the obtuse angle with the Resins, epoxy sealing coat.
5. external-compression type hollow fiber column according to claim 1 is characterized in that: described hollow-fibre membrane bore is the 0.3-0.5 millimeter, and its molecular retention value is 1000-50,000 dalton.
6. external-compression type hollow fiber column according to claim 1 is characterized in that: the material of described hollow fiber film tube is a polysulfone material.
7. external-compression type hollow fiber column according to claim 1 is characterized in that: described tubing string is made by synthetic glass.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 99202751 CN2377250Y (en) | 1999-02-05 | 1999-02-05 | External pressure type hollow fibre column |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 99202751 CN2377250Y (en) | 1999-02-05 | 1999-02-05 | External pressure type hollow fibre column |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN2377250Y true CN2377250Y (en) | 2000-05-10 |
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ID=33998089
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 99202751 Expired - Fee Related CN2377250Y (en) | 1999-02-05 | 1999-02-05 | External pressure type hollow fibre column |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN2377250Y (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107206323A (en) * | 2014-09-22 | 2017-09-26 | 费森尤斯医药用品德国有限公司 | Point of use water purifier with polysulfone hollow fibre |
-
1999
- 1999-02-05 CN CN 99202751 patent/CN2377250Y/en not_active Expired - Fee Related
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107206323A (en) * | 2014-09-22 | 2017-09-26 | 费森尤斯医药用品德国有限公司 | Point of use water purifier with polysulfone hollow fibre |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C56 | Change in the name or address of the patentee |
Owner name: SHANGHAI FUDAN ZHANGJIANG BIOMEDICAL CO., LTD. Free format text: FORMER NAME OR ADDRESS: SHANGHAI FUDAN UNIVERSITY ZHANGJIANG BIOMEDICAL LTD. |
|
| CP01 | Change in the name or title of a patent holder |
Patentee after: Fudan Zhangjiang Biological Medicine Co., Ltd., Shanghai Patentee before: Zhangjiang Biological Medicine Co., Ltd., Shanghai Fudan |
|
| C19 | Lapse of patent right due to non-payment of the annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |