CN107999036A - The high pressure resistant affinity chromatography medium of chromatographic column - Google Patents
The high pressure resistant affinity chromatography medium of chromatographic column Download PDFInfo
- Publication number
- CN107999036A CN107999036A CN201711277799.9A CN201711277799A CN107999036A CN 107999036 A CN107999036 A CN 107999036A CN 201711277799 A CN201711277799 A CN 201711277799A CN 107999036 A CN107999036 A CN 107999036A
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- CN
- China
- Prior art keywords
- medium
- affinity chromatography
- high pressure
- pressure resistant
- chromatographic column
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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- 238000001042 affinity chromatography Methods 0.000 title claims abstract description 27
- 239000012501 chromatography medium Substances 0.000 title claims abstract description 26
- GWEVSGVZZGPLCZ-UHFFFAOYSA-N Titan oxide Chemical compound O=[Ti]=O GWEVSGVZZGPLCZ-UHFFFAOYSA-N 0.000 claims abstract description 42
- 239000002609 medium Substances 0.000 claims abstract description 40
- XLOMVQKBTHCTTD-UHFFFAOYSA-N Zinc monoxide Chemical compound [Zn]=O XLOMVQKBTHCTTD-UHFFFAOYSA-N 0.000 claims abstract description 32
- 239000004408 titanium dioxide Substances 0.000 claims abstract description 21
- 239000011787 zinc oxide Substances 0.000 claims abstract description 16
- 229920001661 Chitosan Polymers 0.000 claims abstract description 10
- 239000004425 Makrolon Substances 0.000 claims abstract description 8
- 239000004005 microsphere Substances 0.000 claims abstract description 8
- 229920000515 polycarbonate Polymers 0.000 claims abstract description 8
- 125000002887 hydroxy group Chemical group [H]O* 0.000 claims abstract description 4
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 claims 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims 2
- 229910052760 oxygen Inorganic materials 0.000 claims 2
- 239000001301 oxygen Substances 0.000 claims 2
- 239000011701 zinc Substances 0.000 claims 2
- 229910052725 zinc Inorganic materials 0.000 claims 2
- 238000004519 manufacturing process Methods 0.000 abstract description 4
- 239000002245 particle Substances 0.000 abstract description 3
- 238000000034 method Methods 0.000 abstract description 2
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 20
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 12
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 8
- 238000001035 drying Methods 0.000 description 8
- 239000002904 solvent Substances 0.000 description 8
- 238000003756 stirring Methods 0.000 description 8
- 239000003814 drug Substances 0.000 description 5
- LSTRKXWIZZZYAS-UHFFFAOYSA-N 2-bromoacetyl bromide Chemical class BrCC(Br)=O LSTRKXWIZZZYAS-UHFFFAOYSA-N 0.000 description 4
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 4
- BVKZGUZCCUSVTD-UHFFFAOYSA-N carbonic acid Chemical compound OC(O)=O BVKZGUZCCUSVTD-UHFFFAOYSA-N 0.000 description 4
- 239000000969 carrier Substances 0.000 description 4
- 150000002148 esters Chemical class 0.000 description 4
- 238000001914 filtration Methods 0.000 description 4
- 150000004679 hydroxides Chemical class 0.000 description 4
- 238000002156 mixing Methods 0.000 description 4
- 239000003960 organic solvent Substances 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 229910052708 sodium Inorganic materials 0.000 description 4
- 239000011734 sodium Substances 0.000 description 4
- 238000005406 washing Methods 0.000 description 4
- 229940079593 drug Drugs 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 229920000936 Agarose Polymers 0.000 description 1
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 1
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 230000008676 import Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
Classifications
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/22—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising organic material
- B01J20/26—Synthetic macromolecular compounds
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01D—SEPARATION
- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/26—Selective adsorption, e.g. chromatography characterised by the separation mechanism
- B01D15/38—Selective adsorption, e.g. chromatography characterised by the separation mechanism involving specific interaction not covered by one or more of groups B01D15/265 and B01D15/30 - B01D15/36, e.g. affinity, ligand exchange or chiral chromatography
- B01D15/3804—Affinity chromatography
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/02—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising inorganic material
- B01J20/06—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising inorganic material comprising oxides or hydroxides of metals not provided for in group B01J20/04
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J20/00—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof
- B01J20/22—Solid sorbent compositions or filter aid compositions; Sorbents for chromatography; Processes for preparing, regenerating or reactivating thereof comprising organic material
- B01J20/24—Naturally occurring macromolecular compounds, e.g. humic acids or their derivatives
Landscapes
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Inorganic Chemistry (AREA)
- Solid-Sorbent Or Filter-Aiding Compositions (AREA)
Abstract
This case is related to a kind of high pressure resistant affinity chromatography medium of chromatographic column, including medium one and medium two；Wherein, the medium one is bonded with chitosan molecule using makrolon microballoon as kernel in the makrolon microsphere surface；Hydroxyl terminal connection aglucon in the chitosan molecule；The medium two includes titanium dioxide and zinc oxide.This case introduces high pressure resistant particle by being designed improvement to the formula of affinity chromatography medium in conventional single medium, to improve the mechanical strength of affinity chromatography medium, so as to improve the separative efficiency of chromatography media at elevated pressures and recycle number；Meanwhile this improved method is simple, production cost is low.
Description
Technical field
The present invention relates to a kind of chromatography media, more particularly to a kind of high pressure resistant affinity chromatography medium of chromatographic column.
Background technology
At present, in pharmaceutical field, the purity and biological safety of medicine how is improved, is the biologies such as recombinant protein, antibody
Medicine is researched and developed and the final steps in production process.Acquisition of medicine enterprise in China for the separating medium of chromatographic column rely primarily on into
Mouthful, therefore, state's domestic demand breaks away from the dependence to import separating medium, reduces the production cost of medicine.To protein drug purifies and separates
Affinity chromatography medium is mainly agar sugar product, but this kind of product mechanical strength is relatively low, so as to cause it to be chromatographed in filling
After column, proof pressure reduces, and limits the room for promotion of its separative efficiency, while also inhibits the reuse time of agarose
Number.
The content of the invention
For technical problem existing in the prior art, this case provides a kind of high pressure resistant affinity chromatography medium of chromatographic column.
To achieve the above object, this case is achieved through the following technical solutions：
A kind of high pressure resistant affinity chromatography medium of chromatographic column, wherein, including medium one and medium two；
Wherein, the medium one is bonded with shell in the makrolon microsphere surface and gathers using makrolon microballoon as kernel
Glycan molecule；Hydroxyl terminal connection aglucon in the chitosan molecule；
The medium two includes titanium dioxide and zinc oxide.
Preferably, the high pressure resistant affinity chromatography medium of the chromatographic column, wherein, the matter of the medium one and medium two
Amount is than being 100:12-18.
Preferably, the high pressure resistant affinity chromatography medium of the chromatographic column, wherein, the matter of the medium one and medium two
Amount is than being 100:14-16.
Preferably, the high pressure resistant affinity chromatography medium of the chromatographic column, wherein, the titanium dioxide and zinc oxide
Mass ratio is 10:6-8.
Preferably, the high pressure resistant affinity chromatography medium of the chromatographic column, wherein, the titanium dioxide and zinc oxide
Mass ratio is 10:7.
Preferably, the high pressure resistant affinity chromatography medium of the chromatographic column, wherein, the particle diameter of the titanium dioxide is
10-20nm。
The beneficial effects of the invention are as follows：
This case introduces resistance to height by being designed improvement to the formula of affinity chromatography medium in conventional single medium
Grain son, to improve the mechanical strength of affinity chromatography medium, thus improve the separative efficiency of chromatography media at elevated pressures and
Recycle number；Meanwhile this improved method is simple, production cost is low.
Embodiment
With reference to embodiment, the present invention is described in further detail, to make those skilled in the art with reference to specification
Word can be implemented according to this.
A kind of high pressure resistant affinity chromatography medium of chromatographic column, wherein, including medium one and medium two；
Wherein, the medium one is bonded with shell in the makrolon microsphere surface and gathers using makrolon microballoon as kernel
Glycan molecule；Hydroxyl terminal connection aglucon in the chitosan molecule；
The medium two includes titanium dioxide and zinc oxide.
Wherein, the mass ratio of the medium one and medium two is 100:12-18.
Wherein, the mass ratio of the medium one and medium two is 100:14-16.
Wherein, the mass ratio of the titanium dioxide and zinc oxide is 10:6-8.
Wherein, the mass ratio of the titanium dioxide and zinc oxide is 10:7.
Wherein, the particle diameter of the titanium dioxide is 10-20nm.
Embodiment 1
Preparation process is as follows：
1) 5-8mol dimethyl sulfoxide (DMSO)s, 1.2-1.5mol bromoacetyl bromides and 0.5-1mol hydroxides are added into 1mol carriers
Sodium, stirs 3h at 25 DEG C, and after reaction, filtering, washs drying；Carrier has the poly- carbonic acid of chitosan molecule for surface bond
Ester microsphere；
2) aglucon, the acetone of 8mol and the sodium hydroxide of 4mol of 1.5mol is added, stirring reaction 8h, reaction at 45 DEG C
After be filtered to remove solvent, washing is three times；
3) it is 10 by the mass ratio of medium one and medium two:13rd, the mass ratio of titanium dioxide and zinc oxide is 10:6 ratio
Example adds titanium dioxide and zinc oxide, and mixing is sufficiently stirred under organic solvent, filters off solvent, is drying to obtain affinity chromatography Jie
Matter.
Embodiment 2
Preparation process is as follows：
1) 5-8mol dimethyl sulfoxide (DMSO)s, 1.2-1.5mol bromoacetyl bromides and 0.5-1mol hydroxides are added into 1mol carriers
Sodium, stirs 3h at 25 DEG C, and after reaction, filtering, washs drying；Carrier has the poly- carbonic acid of chitosan molecule for surface bond
Ester microsphere；
2) aglucon, the acetone of 8mol and the sodium hydroxide of 4mol of 1.5mol is added, stirring reaction 8h, reaction at 45 DEG C
After be filtered to remove solvent, washing is three times；
3) it is 10 by the mass ratio of medium one and medium two:15th, the mass ratio of titanium dioxide and zinc oxide is 10:7 ratio
Example adds titanium dioxide and zinc oxide, and mixing is sufficiently stirred under organic solvent, filters off solvent, is drying to obtain affinity chromatography Jie
Matter.
Embodiment 3
Preparation process is as follows：
1) 5-8mol dimethyl sulfoxide (DMSO)s, 1.2-1.5mol bromoacetyl bromides and 0.5-1mol hydroxides are added into 1mol carriers
Sodium, stirs 3h at 25 DEG C, and after reaction, filtering, washs drying；Carrier has the poly- carbonic acid of chitosan molecule for surface bond
Ester microsphere；
2) aglucon, the acetone of 8mol and the sodium hydroxide of 4mol of 1.5mol is added, stirring reaction 8h, reaction at 45 DEG C
After be filtered to remove solvent, washing is three times；
3) it is 10 by the mass ratio of medium one and medium two:16th, the mass ratio of titanium dioxide and zinc oxide is 10:8 ratio
Example adds titanium dioxide and zinc oxide, and mixing is sufficiently stirred under organic solvent, filters off solvent, is drying to obtain affinity chromatography Jie
Matter.
Embodiment 4
Preparation process is as follows：
1) 5-8mol dimethyl sulfoxide (DMSO)s, 1.2-1.5mol bromoacetyl bromides and 0.5-1mol hydroxides are added into 1mol carriers
Sodium, stirs 3h at 25 DEG C, and after reaction, filtering, washs drying；Carrier has the poly- carbonic acid of chitosan molecule for surface bond
Ester microsphere；
2) aglucon, the acetone of 8mol and the sodium hydroxide of 4mol of 1.5mol is added, stirring reaction 8h, reaction at 45 DEG C
After be filtered to remove solvent, washing is three times；
3) it is 10 by the mass ratio of medium one and medium two:18th, the mass ratio of titanium dioxide and zinc oxide is 10:6 ratio
Example adds titanium dioxide and zinc oxide, and mixing is sufficiently stirred under organic solvent, filters off solvent, is drying to obtain affinity chromatography Jie
Matter.
Following table lists the test result of embodiment 1-4：
Although embodiment of the present invention is disclosed as above, it is not restricted in specification and embodiment listed
With it can be applied to various suitable the field of the invention completely, can be easily for those skilled in the art
Realize other modification, therefore under the universal limited without departing substantially from claim and equivalency range, it is of the invention and unlimited
In specific details.
Claims (6)
1. the high pressure resistant affinity chromatography medium of a kind of chromatographic column, it is characterised in that including medium one and medium two；
Wherein, the medium one is bonded with chitosan point using makrolon microballoon as kernel in the makrolon microsphere surface
Son；Hydroxyl terminal connection aglucon in the chitosan molecule；
The medium two includes titanium dioxide and zinc oxide.
2. the high pressure resistant affinity chromatography medium of chromatographic column as claimed in claim 1, it is characterised in that the medium one and medium
Two mass ratio is 100:12-18.
3. the high pressure resistant affinity chromatography medium of chromatographic column as claimed in claim 1, it is characterised in that the medium one and medium
Two mass ratio is 100:14-16.
4. the high pressure resistant affinity chromatography medium of chromatographic column as claimed in claim 1, it is characterised in that the titanium dioxide and oxygen
The mass ratio for changing zinc is 10:6-8.
5. the high pressure resistant affinity chromatography medium of chromatographic column as claimed in claim 1, it is characterised in that the titanium dioxide and oxygen
The mass ratio for changing zinc is 10:7.
6. the high pressure resistant affinity chromatography medium of chromatographic column as claimed in claim 1, it is characterised in that the grain of the titanium dioxide
Footpath is 10-20nm.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711277799.9A CN107999036A (en) | 2017-12-06 | 2017-12-06 | The high pressure resistant affinity chromatography medium of chromatographic column |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711277799.9A CN107999036A (en) | 2017-12-06 | 2017-12-06 | The high pressure resistant affinity chromatography medium of chromatographic column |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107999036A true CN107999036A (en) | 2018-05-08 |
Family
ID=62057041
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711277799.9A Withdrawn CN107999036A (en) | 2017-12-06 | 2017-12-06 | The high pressure resistant affinity chromatography medium of chromatographic column |
Country Status (1)
| Country | Link |
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| CN (1) | CN107999036A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114054006A (en) * | 2021-11-17 | 2022-02-18 | 苏州纳微科技股份有限公司 | Oligo-dT affinity chromatography filler |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110217539A1 (en) * | 2009-02-19 | 2011-09-08 | Alex Garfield Bonner | Porous interpenetrating polymer network |
| WO2012030512A1 (en) * | 2010-09-03 | 2012-03-08 | Percivia Llc. | Flow-through protein purification process |
| CN106475074A (en) * | 2016-12-22 | 2017-03-08 | 苏州楚博生物技术有限公司 | High mechanical properties affinity chromatography medium |
| CN106582580A (en) * | 2016-12-22 | 2017-04-26 | 苏州楚博生物技术有限公司 | High-pressure-resistant affinity chromatography media for chromatographic column |
-
2017
- 2017-12-06 CN CN201711277799.9A patent/CN107999036A/en not_active Withdrawn
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110217539A1 (en) * | 2009-02-19 | 2011-09-08 | Alex Garfield Bonner | Porous interpenetrating polymer network |
| WO2012030512A1 (en) * | 2010-09-03 | 2012-03-08 | Percivia Llc. | Flow-through protein purification process |
| CN106475074A (en) * | 2016-12-22 | 2017-03-08 | 苏州楚博生物技术有限公司 | High mechanical properties affinity chromatography medium |
| CN106582580A (en) * | 2016-12-22 | 2017-04-26 | 苏州楚博生物技术有限公司 | High-pressure-resistant affinity chromatography media for chromatographic column |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114054006A (en) * | 2021-11-17 | 2022-02-18 | 苏州纳微科技股份有限公司 | Oligo-dT affinity chromatography filler |
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| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20180508 |
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