CN101690854B - Novel expanded bed - Google Patents
Novel expanded bed Download PDFInfo
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- CN101690854B CN101690854B CN2009101768351A CN200910176835A CN101690854B CN 101690854 B CN101690854 B CN 101690854B CN 2009101768351 A CN2009101768351 A CN 2009101768351A CN 200910176835 A CN200910176835 A CN 200910176835A CN 101690854 B CN101690854 B CN 101690854B
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- expansion bed
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- 239000012530 fluid Substances 0.000 claims abstract description 13
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- 238000001179 sorption measurement Methods 0.000 claims description 4
- 229920005479 Lucite® Polymers 0.000 claims description 2
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- 239000004926 polymethyl methacrylate Substances 0.000 claims description 2
- 239000010935 stainless steel Substances 0.000 claims 1
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- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 8
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- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
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- Treatment Of Liquids With Adsorbents In General (AREA)
Abstract
The invention relates to a expanded bed, which comprises an expanded bed post pipe, an inner pipe fixing device and a fluid distribution device, wherein the expanded bed post pipe comprises an outer pipe and an inner pipe which are coaxial and are arranged in the center of the post pipe; and the inner pipe is fixed in the post pipe by the inner pipe fixing device. By fixing the inner pipe in the expanded bed post pipe, the migration of a mobile phase in an expanded bed layer is changed, the back-mixing phenomenon in the expanded bed is reduced, and the separation efficiency of the expanded bed is improved. The novel expanded bed has quite high practical value.
Description
Technical field
The present invention relates to the expansion bed technical field, be specifically related to a kind of expansion bed of structure.
Background technology
People such as Chase had installed a kind of special fluid distributor in traditional chromatographic column bottom in 1992, obtain a stable expansion bed with traditional agarose chromatography medium, adopt this covering device to handle the protein and the cell mixture that can contain solid particle, its chromatographic property approaches the packed bed chromatography, thereby develop a kind of novel living beings unit removal techniques---Expanded Bed Adsorption chromatography (Expanded bed adsorption chromatograph is called for short EBA).Expansion bed also can be interpreted as the very little fluid bed of back-mixing, is a kind of special shape of fluid bed.Expansion bed combines the advantage of fluid bed and packed bed chromatography, different with the mode of operation of traditional chromatography, the expansion bed feed liquid is that the bottom from bed enters, because speed and buoyancy that fluid upwards flows and produces, cause the adsorbent of interior different size of bed or density to exist upwards unsteady to some extent, cause bed expansion, after treating that bed is stable, different adsorbents can be on the fixed position separately in the expansion bed more among a small circle in fluctuate, axial mixability between the solid-liquid phase is lower, fluid is with the bed of flowing through near the mode of piston flow, target substance in the feed liquid is attracted on the adsorbent by the pattern of packed bed basically, thereby obtains good separating effect.Because the expansion of bed, voidage increases between the adsorbent, can make small solid particles such as cell in the feed liquid, cell fragment pass through bed smoothly, therefore, ExPANDED BED ADSORPTION TECHNIQUE is a kind of integrated bio lock out operation technology, it integrates clarification, concentrates and the initial gross separation purifying, can directly from the material liquid that contains particle, adsorb and the preliminary purification target substance, its handle clarifies, concentrates and the preliminary purification several steps is integrated in the unit operations, reduced operating procedure, improve product yield, reduced purifying expense and capital input.Therefore, this technology is subjected to bio-separation purification technique expert's attention once coming out.At present, expansion bed has been successfully used to the extraction of target product in Escherichia coli, yeast, animal cell culture liquid and animal tissue, plant tissue, fruit juice, animal milk and some other microorganisms, also can be with the checkout gear of expansion bed as bioreactor and biological substance.
The expansion bed technology has tangible characteristics and advantage, but because the bands of a spectrum broadening effect of expansion bed inside, having radial and axial back-mixing to a certain degree in the bed is the one of the main reasons that influences its separation accuracy; Simultaneously, the adsorbent kind of having developed that is applicable to expansion bed is few, makes the expansion bed technology still can't substitute the leading position of traditional chromatographic technique in the bio-separation field.At present, mainly concentrate on the development of adsorbent and use, as the research of the high density core-shell type adsorbent of inventions such as Sun Yan at the research of expansion bed aspect; Su Zhiguo etc. utilize the expansion bed technology to separate bovine serum albumin(BSA) from ox blood, only through single stepping, obtain purity near 70% monomer bovine serum albumin(BSA) (BSA), have realized separating effectively fast of BSA.Anita Pai etc. prepares a kind of hard matrix of cross-linked cellulose of super big hole structure, pore radius reaches 3 μ m, matrix is kept very low HETP (HETP) and higher adsorption capacity in very big flow rates, can realize the expansion bed mode wash-out under the high flow rate equally.At the research of expansion bed configuration aspects report seldom, particularly about the expansion bed rod structure.The expansion column is the main composition of expansion bed technology, starts with from rod structure, considers to weaken the radially back-mixing in the expansion bed bed, and improving separative efficiency should be a kind of new method.
Summary of the invention
The object of the present invention is to provide a kind of expansion bed technology separation accuracy that can solve to hang down and inefficient expansion bed.
Expansion bed of the present invention, comprise expansion bed column jecket (interior pipe and outer tube), interior tube fixer, fluid distributor, bed height adjuster, column cap, sieve plate, its middle external tube is common gc column tube, and interior pipe size is littler than the outer tube, the coaxial outer tube hub of expansion bed that places of interior pipe.
The above expands column, and described interior pipe can be fixed in the expansion bed column jecket by fixator, and interior pipe size is littler than outer tube, can assemble or dismantle.
The above expands column, and described outer tube and interior pipe are ganoid glass tube or lucite tube.
The above expansion bed, fixator is cross or circular for the band groove, and the size of fixator equates that with the internal diameter size of expansion bed outer tube the size of groove equates that with the inner tube diameter size pipe and fixator removably connect in the chromatographic column.
The above expansion bed, interior tube fixer are high-molecular organic material.
The above expansion bed, the chromatogram column cap at gc column tube and two ends is for removably connecting.
The above expansion bed, chromatogram column cap are provided with standard colour chart post interface.
The above expands column, and the gap between described interior pipe and the outer tube is between 10-500mm, between the radially ratio 0.1-0.8 of interior pipe and outer tube.
The above expands column, and described interior pipe and outer tube axial scale are between the 0.1-0.9.
Another object of the present invention is to provides a kind of analytical method to the research of the key property of expansion bed.
In the analytical method of this expansion bed, the size of pipe in can selecting neatly as the case may be.Used filler is commercialization medium streamline-DEAE in the analytic process, is filled in to uniform filling column jecket.Analytic process need be examined or check different interior pipe sizes to the expansion bed Effect on Performance.
The present invention is by the design of pipe and outer tube in expansion bed, changed mobile flow trace in the expansion bed, weakened the back-mixing problem of mobile phase, thereby improved the separative efficiency of expansion bed, have very high practical value, be with a wide range of applications in downstream technical fields such as biology, chemical industry, medicine.
Description of drawings
Fig. 1: expansion bed chromatographic column of the present invention and traditional expansion bed chromatographic column schematic cross-section
Fig. 2: expansion bed chromatographic column generalized section of the present invention
Fig. 3: traditional expansion bed chromatographic column generalized section
Fig. 4: expansion bed chromatographic column of the present invention detects collection of illustrative plates (interior pipe is 60mm)
Fig. 5: traditional expansion bed chromatographic column detects collection of illustrative plates
The specific embodiment
The present invention has designed a kind of expansion bed, by the combination that interior pipe and outer tube organically combine by a certain percentage, influences the motion of the phase that flows in the bed, has weakened the air-teturning mixed phenomenon of the phase that flows in the bed, has improved the separative efficiency of expansion bed chromatographic column.
Below the structure of the expansion bed of the present invention effect forming and produced just, cooperate instantiation to be described in detail as follows:
Shown in Figure 2 is expansion bed chromatographic column generalized section of the present invention, and Fig. 3 is traditional expansion bed chromatographic column generalized section.
Example 1
With internal diameter is 26mm, length is that the lower end column cap of traditional expansion bed chromatographic column of 260mm is turned on (being the fluid distributor end), with internal diameter is 13mm, length is that the interior pipe of 60mm is fixed in the expansion bed outer tube (being fixed by interior tube fixer), interior pipe places the column jecket center with outer coaxial tube, and the 100mm place (interior pipe lower end is 40mm apart from fluid distributor) of column jecket is fixed in interior pipe upper end.The column cap of the chromatographic column of screwing on lower end.With commercialization medium streamline-DEAE is filled media, slowly pours into from the gc column tube upper end, and the medium packed height reaches column jecket 75mm place.Regulate the position of the bed height adjuster of expansion bed then, be positioned at pipe upper end (being the 100mm place), level altitude adjuster and column cap.After the expansion bed chromatographic column assembled, the tube connector of expansion bed chromatographic column lower end and upper end linked to each other with inlet with the outlet of detector respectively.This tests used detector is AKTA Explore 100 fast separating and purifying systems (Amersham Biosciences companies).The analyzing and testing condition: flowing is ultra-pure water mutually, and analytic sample is 10% acetone soln, and sample size is 0.5ml, and the detection wavelength is 280nm.Operating procedure: flowing enters from expansion bed gc column tube lower end, regulates flow velocity and makes bed expansion, and when flow velocity is 67.8cm/h, bed expansion is to the 100mm place, balance 20-30min, the acetone of pulse sample introduction 0.5ml, log.Under similarity condition, in contrast with the measure-alike traditional chromatographic column of outer tube.Experimental result is as follows:
Table 1 chromatographic parameter
| Peak height mAU | Half-peak breadth (ml) | Theoretical cam curve | Symmetrical factor | |
| Common expansion bed | 45.987 | 26.400 | 6.630 | 3.550 |
| Expansion bed | 55.406 | 24.129 | 16.432 | 1.220 |
Result of the test shows: under the same conditions, the chromatographic performance of having fixed the expansion bed of pipe in the 60mm will be significantly better than unidimensional common expansion bed, peak type symmetry, and it is many that theoretical cam curve increases twice than common expansion bed, the separative efficiency height.
Example 2
With internal diameter is 26mm, length is that the lower end column cap of traditional expansion bed chromatographic column of 260mm is turned on (being the fluid distributor end), with internal diameter is 13mm, length is that the interior pipe of 40mm is fixed in the expansion bed outer tube (being fixed by interior tube fixer), interior pipe places the column jecket center with outer coaxial tube, and the 100mm place (interior pipe lower end is 60mm apart from fluid distributor) of column jecket is fixed in interior pipe upper end.The column cap of the chromatographic column of screwing on lower end.With commercialization medium streamline-DEAE is filled media, slowly pours into from the gc column tube upper end, and the medium packed height reaches column jecket 75mm place.Regulate the position of the bed height adjuster of expansion bed then, be positioned at pipe upper end (being the 100mm place), level altitude adjuster and column cap.After the expansion bed chromatographic column assembled, the tube connector of expansion bed chromatographic column lower end and upper end linked to each other with inlet with the outlet of detector respectively.This tests used detector is AKTA Explore 100 fast separating and purifying systems (Amersham Biosciences companies).The analyzing and testing condition: flowing is ultra-pure water mutually, and analytic sample is 10% acetone soln, and sample size is 0.5ml, and the detection wavelength is 280nm.Operating procedure: flowing enters from expansion bed gc column tube lower end, regulates flow velocity and makes bed expansion, and when flow velocity is 62.2cm/h, bed expansion is to the 100mm place, balance 20-30min, the acetone of pulse sample introduction 0.5ml, log.Under similarity condition, in contrast with the measure-alike traditional chromatographic column of outer tube.By parameters such as the theory of computation numbers of plates, result of the test shows: under the same conditions, the chromatographic performance of having fixed the expansion bed of pipe in the 40mm is better than unidimensional common expansion bed, peak type symmetry; Theoretical cam curve is bigger than common expansion bed, the separative efficiency height.
Aforementionedly be a kind of reasonably using method of the present invention, one of the mode that can specifically implement for the present invention only is not as limit.Make not breaking away under the spirit of the present invention, still can change its form and details.
Claims (5)
1. an expansion bed comprises expansion bed column jecket, interior tube fixer and fluid distributor, and wherein the expansion bed column jecket is made up of outer tube and interior pipe, interior pipe and outer coaxial tube; Outer tube diameter is of a size of 100-2000mm, column length 200-6000mm; Diameter of inner pipe is of a size of 20-1600mm, column length 100-4000mm; Interior pipe and outer tube diameter are to than being 0.1-0.8, axially than being 0.1-0.9; Interior pipe is fixed in the outer tube by interior tube fixer, and interior tube fixer is cross or round for having groove, and the size of fixator equates that with the outer tube diameter size groove size equates with the inner tube diameter size.
2. expansion bed as claimed in claim 1 is characterized in that the size of interior pipe is less than outer tube.
3. expansion bed as claimed in claim 1 is characterized in that interior pipe and fixator are detachable.
4. expansion bed as claimed in claim 1 is characterized in that, described column tube material is the material that glass or lucite class do not have specific adsorption, smooth surface.
5. expansion bed as claimed in claim 1 is characterized in that, interior tube fixer is machinable macromolecular material or stainless steel.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009101768351A CN101690854B (en) | 2009-09-22 | 2009-09-22 | Novel expanded bed |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2009101768351A CN101690854B (en) | 2009-09-22 | 2009-09-22 | Novel expanded bed |
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| Publication Number | Publication Date |
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| CN101690854A CN101690854A (en) | 2010-04-07 |
| CN101690854B true CN101690854B (en) | 2011-08-31 |
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| CN2009101768351A Expired - Fee Related CN101690854B (en) | 2009-09-22 | 2009-09-22 | Novel expanded bed |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6620326B1 (en) * | 1998-06-18 | 2003-09-16 | Upfront Chromatography A/S | Expanded bed adsorption system |
| CN2608501Y (en) * | 2003-04-11 | 2004-03-31 | 上海华震科技有限公司 | Expansion bed absorption separator |
| CN1583211A (en) * | 2004-06-07 | 2005-02-23 | 浙江大学宁波理工学院 | Expanding bed apparatus |
| CN101393187A (en) * | 2007-09-21 | 2009-03-25 | 北京化工大学 | Liquid chromatography column and high performance liquid chromatography analytical method |
-
2009
- 2009-09-22 CN CN2009101768351A patent/CN101690854B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6620326B1 (en) * | 1998-06-18 | 2003-09-16 | Upfront Chromatography A/S | Expanded bed adsorption system |
| CN2608501Y (en) * | 2003-04-11 | 2004-03-31 | 上海华震科技有限公司 | Expansion bed absorption separator |
| CN1583211A (en) * | 2004-06-07 | 2005-02-23 | 浙江大学宁波理工学院 | Expanding bed apparatus |
| CN101393187A (en) * | 2007-09-21 | 2009-03-25 | 北京化工大学 | Liquid chromatography column and high performance liquid chromatography analytical method |
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| Publication number | Publication date |
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| CN101690854A (en) | 2010-04-07 |
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Owner name: BEIJING UNIVERSITY OF CHEMICAL TECHNOLOGY Free format text: FORMER OWNER: ZHOU XIN Effective date: 20120206 |
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Effective date of registration: 20120206 Address after: 100029 Chaoyang District, North Third Ring Road,, No. 15, Beijing University of Chemical Technology Patentee after: Beijing University of Chemical Technology Address before: 100029 No. 15 East Third Ring Road, Beijing, Beijing University of Chemical Technology, No. 53, Patentee before: Zhou Xin |
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