CN108796826A - Mixed mode nano fibrous membrane chromatography media and preparation method thereof - Google Patents
Mixed mode nano fibrous membrane chromatography media and preparation method thereof Download PDFInfo
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- CN108796826A CN108796826A CN201810660682.7A CN201810660682A CN108796826A CN 108796826 A CN108796826 A CN 108796826A CN 201810660682 A CN201810660682 A CN 201810660682A CN 108796826 A CN108796826 A CN 108796826A
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- mixed mode
- nano fibrous
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- fibrous membrane
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- D—TEXTILES; PAPER
- D04—BRAIDING; LACE-MAKING; KNITTING; TRIMMINGS; NON-WOVEN FABRICS
- D04H—MAKING TEXTILE FABRICS, e.g. FROM FIBRES OR FILAMENTARY MATERIAL; FABRICS MADE BY SUCH PROCESSES OR APPARATUS, e.g. FELTS, NON-WOVEN FABRICS; COTTON-WOOL; WADDING ; NON-WOVEN FABRICS FROM STAPLE FIBRES, FILAMENTS OR YARNS, BONDED WITH AT LEAST ONE WEB-LIKE MATERIAL DURING THEIR CONSOLIDATION
- D04H1/00—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres
- D04H1/70—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres
- D04H1/72—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres the fibres being randomly arranged
- D04H1/728—Non-woven fabrics formed wholly or mainly of staple fibres or like relatively short fibres characterised by the method of forming fleeces or layers, e.g. reorientation of fibres the fibres being randomly arranged by electro-spinning
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- D—TEXTILES; PAPER
- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01D—MECHANICAL METHODS OR APPARATUS IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS
- D01D5/00—Formation of filaments, threads, or the like
- D01D5/0007—Electro-spinning
- D01D5/0015—Electro-spinning characterised by the initial state of the material
- D01D5/003—Electro-spinning characterised by the initial state of the material the material being a polymer solution or dispersion
-
- D—TEXTILES; PAPER
- D01—NATURAL OR MAN-MADE THREADS OR FIBRES; SPINNING
- D01D—MECHANICAL METHODS OR APPARATUS IN THE MANUFACTURE OF ARTIFICIAL FILAMENTS, THREADS, FIBRES, BRISTLES OR RIBBONS
- D01D5/00—Formation of filaments, threads, or the like
- D01D5/0007—Electro-spinning
- D01D5/0061—Electro-spinning characterised by the electro-spinning apparatus
-
- D—TEXTILES; PAPER
- D10—INDEXING SCHEME ASSOCIATED WITH SUBLASSES OF SECTION D, RELATING TO TEXTILES
- D10B—INDEXING SCHEME ASSOCIATED WITH SUBLASSES OF SECTION D, RELATING TO TEXTILES
- D10B2505/00—Industrial
- D10B2505/04—Filters
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- Engineering & Computer Science (AREA)
- Textile Engineering (AREA)
- Mechanical Engineering (AREA)
- Chemical & Material Sciences (AREA)
- Dispersion Chemistry (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention discloses mixed mode nano fibrous membrane chromatography medias and preparation method thereof, and preparation method is：(1) it by the PVDF and SMA mixings after drying, is dissolved in the DMF after drying and is configured to spinning solution, carry out electrostatic spinning and prepare static spinning membrane, be freeze-dried；(2) film that step (1) obtains is dipped in amine or amine aqueous solution, is impregnated, use deionized water and absolute ethyl alcohol to rinse successively after taking-up, freeze-drying obtains mixed mode nano fibrous membrane chromatography media.There is mixed mode nano fibrous membrane chromatography media prepared by the present invention higher ligand density can reach higher protein adsorption quantity, method of the invention is simple, mild condition in shorter time of equilibrium adsorption.
Description
Technical field
The invention belongs to bio-separation fields, and in particular to a kind of mixed mode nanofiber film layer of high protein adsorbance
Analyse medium and preparation method thereof.
Background technology
Protein articles occupy critically important status in medical industry and food manufacturing industry, and pharmaceutical grade protein is cured in clinic
The targeting and high efficiency being had shown that in make it be expected to substitute traditional chemical small molecule drug.But protein
It purifies costly, occupies its total production cost more than half.Therefore, a kind of protein purification of economical and efficient is developed
Method is of great significance to research fields such as medical diagnosis and pharmaceutical developments.
The chromatography media for being presently used for protein purification is chromatography column packing and chromatography film medium, although the chromatography prepared
Column has excellent separating property, but there are pressure drops in operating process it is big, flow velocity is low the problems such as, and single-mode in document report
The protein adsorption quantity for chromatographing film medium is not high, though Mixed-Modechromatography film medium improves protein adsorption quantity, film-forming method has one
It settles finally sex-limited.
Invention content
The purpose of the present invention is overcoming the shortcomings of the prior art, a kind of mixed mode of high protein adsorbance is provided and is received
Rice tunica fibrosa chromatography media.
Second object of the present invention is to provide a kind of preparation method of mixed mode nano fibrous membrane chromatography media.
Technical scheme of the present invention is summarized as follows：
The preparation method of mixed mode nano fibrous membrane chromatography media, includes the following steps：
(1) in mass ratio it is (6-20)：PVDF and SMA mixings after drying is dissolved in the DMF after drying by 3 ratio
It is configured to the spinning solution that mass concentration is 9%-18%, electrostatic spinning is carried out and prepares static spinning membrane, be freeze-dried；
The PVDF is the abbreviation of polyvinyladine floride；SMA is the abbreviation of alternate styrene-maleic anhydride copolymer；DMF is
The abbreviation of N,N-dimethylformamide；
(2) film that step (1) obtains is dipped in amine or amine aqueous solution, at 30-80 DEG C, impregnates 3-12h, after taking-up successively
It is rinsed with deionized water and absolute ethyl alcohol, freeze-drying obtains mixed mode nano fibrous membrane chromatography media.
The excellent mass ratio of PVDF and SMA after drying is 10:3.
The preferred 320-350KDa of Mw of the Mw preferred 300-320KDa, SMA of PVDF.
Electrostatic spinning solidification distance is preferably 5-20cm.
The fltting speed of electrostatic spinning is preferably 0.001mm/s-0.008mm/s.
Electrostatic spinning voltage is preferably 12kV-21kV.
Amine is preferably n-butylamine, n-octyl amine or N- methyl glucose amine.
Mixed mode nano fibrous membrane chromatography media prepared by the above method.
Advantages of the present invention：
Mixed mode nano fibrous membrane chromatography media prepared by the present invention has higher ligand density, in shorter suction
In attached equilibration time, higher protein adsorption quantity can be reached, method of the invention is simple, mild condition, in Protein Separation side
Face has a extensive future.
Description of the drawings
Fig. 1 is mixed mode nano fibrous membrane chromatography media and (film that step (1) obtains) before modified prepared by embodiment 1
Infrared figure.
Fig. 2 is IgG solution curve of adsorption kinetics of the PVDFS-BAE films to 1g/L.
Fig. 3 is α-CTP solution curve of adsorption kinetics of the PVDFS-OAE films to 1g/L.
Specific implementation mode：
It will be helpful to understand the present invention by following embodiment, but be not intended to limit present disclosure.In addition, it should also be understood that,
After having read the content that the present invention is told about, those skilled in the art can make various modifications or changes to the present invention, these etc.
Valence form equally falls within the range of following claims.
Below by embodiment, the present invention will be further described.
Each embodiment PVDF dryings preferably dry 12h under the conditions of 75 DEG C.
PVDF is the abbreviation of polyvinyladine floride；SMA is the abbreviation of alternate styrene-maleic anhydride copolymer；DMF is N, N-
The abbreviation of dimethylformamide；
Experiment albumen used is commodity.
Embodiment 1
The preparation method of mixed mode nano fibrous membrane chromatography media, includes the following steps：
(1) it is 6 in mass ratio：3 ratio, PVDF (Mw=300KDa) and the SMA (Mw=320KDa) after drying is mixed
It is even, it is dissolved in the spinning solution for being configured to that mass concentration is 9% in the DMF after drying, in solidification distance 10cm, fltting speed
0.001mm/s carries out electrostatic spinning under the spinning parameter of voltage 15kV and prepares static spinning membrane, is freeze-dried (see in Fig. 1
Curve 1)；
(2) by step (1) obtain film be dipped in n-butylamine (BAE), at 30 DEG C, impregnate 8h, spent successively after taking-up from
Sub- water and absolute ethyl alcohol rinse, freeze-drying, and obtaining mixed mode nano fibrous membrane chromatography media, (abbreviation PVDFS-g-BAE film layers are analysed
Medium).(see the curve 2 in Fig. 1)
Embodiment 2
The preparation method of mixed mode nano fibrous membrane chromatography media, includes the following steps：
(1) it is 20 in mass ratio：3 ratio, PVDF (Mw=310KDa) and the SMA (Mw=340KDa) after drying is mixed
It is even, it is dissolved in the spinning solution for being configured to that mass concentration is 12% in the DMF after drying, in solidification distance 20cm, fltting speed
0.008mm/s carries out electrostatic spinning under the spinning parameter of voltage 21kV and prepares static spinning membrane, is freeze-dried；
(2) by step (1) obtain film be dipped in n-octyl amine (OAE), at 30 DEG C, impregnate 12h, spent successively after taking-up from
Sub- water and absolute ethyl alcohol rinse, freeze-drying, and obtaining mixed mode nano fibrous membrane chromatography media, (abbreviation PVDFS-g-OAE film layers are analysed
Medium) (see the curve 3 in Fig. 1).
Embodiment 3
The preparation method of mixed mode nano fibrous membrane chromatography media, includes the following steps：
(1) it is 10 in mass ratio：3 ratio, PVDF (Mw=320KDa) and the SMA (Mw=350KDa) after drying is mixed
It is even, it is dissolved in the spinning solution for being configured to that mass concentration is 18% in the DMF after drying, in solidification distance 5cm, fltting speed
0.003mm/s carries out electrostatic spinning under the spinning parameter of voltage 12kV and prepares static spinning membrane, is freeze-dried；
(2) film that step (1) obtains is dipped in N- methyl glucoses amine (NMG) aqueous solution of 0.2g/mL, at 80 DEG C, leaching
3h is steeped, uses deionized water and absolute ethyl alcohol to rinse successively after taking-up, freeze-drying obtains mixed mode nano fibrous membrane chromatography media
(abbreviation PVDFS-g-NMG films chromatography media) (see the curve 4 in Fig. 1).
Embodiment 4
The albumen test of mixed mode nano fibrous membrane chromatography media：
Wherein：
BSA:Bovine serum albumin(BSA)
IgG：Immunoglobulin G
Lym：Lysozyme
α-CTP：Chymotrypsin
CYT-C：Cromoci
RNase A：Ribalgilase
PPS:Pepsin
Albumen test process：
A concentration of 1g/L of BSA solution, IgG solution and Lym solution, solvent are the PBS buffer solutions of pH=7；
α-CTP solution concentrations are 2g/L, and solvent is the acetonitrile solution that volumetric concentration is 20%；
CYT-C solution, RNase solution As and PPS solution concentrations are 1g/L, and solvent is the acetonitrile water that volumetric concentration is 20%
Solution；
The method that protein concentration detection uses：BCA methods.
Table 1：The absorption of three kinds of albumen of mixed mode nano fibrous membrane chromatography media pair prepared by embodiment 1 at different pH
Test
PVDFS-g-BAE films chromatography media, in 25mg/ml or more, reaches as high as 42mg/mL to the adsorbance of BSA；
PVDFS-g-BAE films chromatography media, in 50mg/ml or more, reaches as high as 150mg/mL to the adsorbance of IgG；
PVDFS-g-BAE films chromatography media, in 41mg/ml or more, reaches as high as 93mg/mL to the adsorbance of Lym；
Table 2：The absorption of two kinds of albumen of mixed mode nano fibrous membrane chromatography media pair prepared by embodiment 2 at different pH
Test
PVDFS-g-OAE films chromatography media, in 72mg/ml or more, reaches as high as 140mg/mL to the adsorbance of α-CTP；
PVDFS-g-OAE films chromatography media, in 7mg/ml or more, reaches as high as 20mg/mL to the adsorbance of CYT-C；
Table 3：The absorption of two kinds of albumen of mixed mode nano fibrous membrane chromatography media pair prepared by embodiment 3 at different pH
Test
PVDFS-g-NMG films chromatography media, in 26mg/ml or more, reaches as high as 32mg/mL to the adsorbance of RNase A；
PVDFS-g-NMG films chromatography media, in 32mg/ml or more, reaches as high as 93mg/mL to the adsorbance of PPS.
Claims (8)
1. the preparation method of mixed mode nano fibrous membrane chromatography media, it is characterized in that including the following steps：
(1) in mass ratio it is (6-20)：PVDF and SMA mixings after drying is dissolved in the DMF after drying and being prepared by 3 ratio
At the spinning solution that mass concentration is 9%-18%, carries out electrostatic spinning and prepare static spinning membrane, be freeze-dried；
The PVDF is the abbreviation of polyvinyladine floride；SMA is the abbreviation of alternate styrene-maleic anhydride copolymer；DMF is N, N-
The abbreviation of dimethylformamide；
(2) film that step (1) obtains is dipped in amine or amine aqueous solution, at 30-80 DEG C, impregnates 3-12, spent successively after taking-up
Ionized water and absolute ethyl alcohol rinse, and freeze-drying obtains mixed mode nano fibrous membrane chromatography media.
2. according to the method described in claim 1, it is characterized in that the mass ratio of the PVDF and SMA after dry are 10:3.
3. method according to claim 1 or 2, it is characterized in that the Mw of the PVDF is 300-320KDa, the Mw of the SMA
For 320-350KDa.
4. according to the method described in claim 1, it is characterized in that electrostatic spinning solidification distance is 5-20cm.
5. according to the method described in claim 1, it is characterized in that the fltting speed of electrostatic spinning is 0.001mm/s-0.008mm/
s。
6. according to the method described in claim 1, it is characterized in that electrostatic spinning voltage is 12kV-21kV.
7. according to the method described in claim 1, it is characterized in that the amine is n-butylamine, n-octyl amine or N- methyl glucose amine.
8. mixed mode nano fibrous membrane chromatography media prepared by the method for one of claim 1-7.
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| CN201810660682.7A CN108796826A (en) | 2018-06-25 | 2018-06-25 | Mixed mode nano fibrous membrane chromatography media and preparation method thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109316981A (en) * | 2018-12-10 | 2019-02-12 | 天津工业大学 | A kind of preparation method of the super hydrophilic polymer film with demulsification function |
Citations (4)
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|---|---|---|---|---|
| CN101069822A (en) * | 2007-03-06 | 2007-11-14 | 天津大学 | Method for preparing PVDF super-fine fiber film with good mechanics-performance |
| CN103357277A (en) * | 2013-07-24 | 2013-10-23 | 浙江师范大学 | Ultrafiltration membrane with heavy metal ion adsorption function, and preparation method thereof |
| CN105040277A (en) * | 2015-06-26 | 2015-11-11 | 北京理工大学 | Nano-cellulose/cellulose triacetate composite nano-fiber membrane capable of adsorbing and desorbing proteins |
| CN107413312A (en) * | 2017-07-18 | 2017-12-01 | 天津工业大学 | A kind of mixed mode protein adsorption film and preparation method for antibody purification |
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2018
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Patent Citations (4)
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|---|---|---|---|---|
| CN101069822A (en) * | 2007-03-06 | 2007-11-14 | 天津大学 | Method for preparing PVDF super-fine fiber film with good mechanics-performance |
| CN103357277A (en) * | 2013-07-24 | 2013-10-23 | 浙江师范大学 | Ultrafiltration membrane with heavy metal ion adsorption function, and preparation method thereof |
| CN105040277A (en) * | 2015-06-26 | 2015-11-11 | 北京理工大学 | Nano-cellulose/cellulose triacetate composite nano-fiber membrane capable of adsorbing and desorbing proteins |
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| 王栩彬等: "《纳米纤维膜表面修饰制备混合模式亲和膜》", 《第21届全国色谱学术报告会及仪器展览会》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109316981A (en) * | 2018-12-10 | 2019-02-12 | 天津工业大学 | A kind of preparation method of the super hydrophilic polymer film with demulsification function |
| CN109316981B (en) * | 2018-12-10 | 2021-03-09 | 天津工业大学 | Preparation method of super-hydrophilic polymer membrane with demulsification function |
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Application publication date: 20181113 |
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