CN102068965B - Method for preparing chitosan separation medium suitable for protein purification - Google Patents
Method for preparing chitosan separation medium suitable for protein purification Download PDFInfo
- Publication number
- CN102068965B CN102068965B CN 201010579423 CN201010579423A CN102068965B CN 102068965 B CN102068965 B CN 102068965B CN 201010579423 CN201010579423 CN 201010579423 CN 201010579423 A CN201010579423 A CN 201010579423A CN 102068965 B CN102068965 B CN 102068965B
- Authority
- CN
- China
- Prior art keywords
- shitosan
- chitosan
- preparation
- skeleton
- solution
- 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.)
- Active
Links
Images
Abstract
The invention relates to a method for preparing a chitosan separation medium suitable for protein purification, which comprises the following steps of: (1) dispersing acetic acid solution of chitosan into liquid paraffin with stirring to form chitosan particles in the presence of cyclohexane serving as a porogen and a small amount of span80, and further cross-linking the chitosan particles to form a chitosan skeleton under the action of glutaraldehyde serving as a cross-linking agent; (2) swelling the chitosan skeleton, and reacting a hydroxyl group on the chitosan skeleton with epoxy chloropropane in dimethyl sulfoxide (DMSO)/NaOH mixed solution to introduce an epoxy group into the chitosan skeleton so as to obtain a grafted chitosan skeleton; and (3) adding iminodiacetic acid (IDA)/NaOH mixed solution into the grafted chitosan skeleton, reacting at the temperature of between 20 and 80 DEG C for 1 to 10 hours, and performing suction-filtration to obtain the chitosan separation medium. The chitosan separation medium solves the leakage problem of metal ions to a large extent, has the advantages of process stability, high repeatability and the like, and is suitable for mass production.
Description
One. technical field
The invention belongs to the natural macromolecular material technical field, be specifically related to a kind of preparation method who is suitable for the shitosan separating medium of protein purification.
Two. background technology
Although it is a lot of to be used at present the method for Protein Separation, but metal chelate chromatography (IMAC) have chelating media prepare simple and convenient, the exchange carrying capacity is large, separation condition is gentle, highly versatile, be easy to amplify, particularly in the purge process of albumen, the elution requirement that it is gentle, the advantages such as BA that can keep preferably albumen, make its application more and more come into one's own, will become the chromatography method of tool potentiality in protein separation.
But conventional metal chelate chromatography (IMAC) chromatographic column is made carrier mainly with soft matrix such as glucan or agaroses greatly, and these media mostly are import on the one hand, and are expensive, and its technology of preparing is all monopolized by more external biological products major companies; On the other hand, these media in use metal ion easily leak, and cause the decline of Protein Separation purity.Thereby Chinese scholars is being sought inexpensive, good hydrophilic property, IMAC carrier that rigidity is strong always.Shitosan is the unique natural alkaline polysaccharide of finding up to now, and structure and performance and agarose, glucan are similar, aboundresources, and biocompatibility is better, contains active amine (NH in its molecule
2) and hydroxyl (OH), be easy to the porous material that derivatization obtains having various different absorption properties, can direct or modified rear carrier as various chromatographies and protein separation.
The shitosan major part of native state is Powdered, and specific area is little, as absorption carrier, makes carrier and adsorbate all be difficult to reclaim, thereby limits its application.Therefore, shitosan is prepared into monodispersed narrow distribution polymer microsphere, the function of shitosan and polymer microsphere is consistent, make it obtain larger application in fields such as biomedicines.
In recent years, the existing bibliographical information of research about the poly-film of porous shell and microballoon, the shitosan research and development is rapid, but most researchs concentrate on delivery systems such as chitosan modified liposome, microballoon, micro-capsules, and its report as separating medium is also few, and the shitosan separating medium conditional instability of preparation, and adsorb the problems such as carrying capacity is not high.
Three. summary of the invention
The technical problem to be solved in the present invention is to provide a kind of preparation method who is suitable for the shitosan separating medium of protein purification.
For solving the problems of the technologies described above, the present invention adopts following technical scheme:
A kind of preparation method who is suitable for the shitosan separating medium of protein purification comprises the steps:
(1) preparation of shitosan skeleton: under agitation, the acetic acid solution of shitosan is scattered in atoleine, under perforating agent cyclohexane and a little span 80 exist, forms chitosan particle; Then under the effect of crosslinking agent glutaraldehyde, chitosan particle further is cross-linked to form the shitosan skeleton;
(2) chitosan graft: the advanced row of shitosan skeleton swelling treatment; Then in the DMSO/NaOH mixed liquor, the hydroxyl on the shitosan skeleton and epichlorohydrin reaction are introduced epoxy radicals on the shitosan skeleton, obtain the shitosan skeleton after grafting;
(3) preparation of shitosan separating medium: the IDA/NaOH mixed solution is added shitosan skeleton after grafting, react 1～10h at 20～80 ℃ of temperature, suction filtration namely gets described shitosan separating medium; In described IDA/NaOH mixed solution, IDA concentration is 0.5～3.0mol/L, and NaOH concentration is 0.5～3.0mol/L.Wherein IDA is iminodiacetic acid.
Step of the present invention (1) utilizes the via Inverse-Phase Suspension Polymerization preparation to have the shitosan skeleton of gel pore, pearlitic texture.Further, the present invention specifically recommends described step (1) to carry out according to following: it is that in 1.0～3.0% acetum, preparation obtains the chitosan solution that concentration is 0.01～0.05g/mL that shitosan is dissolved in mass fraction; Add successively atoleine (oil phase), cyclohexane and a little Span 80 in reaction vessel, stir the chitosan solution (water) that adds preparation to obtain after 30～60min, be heated to 20～80 ℃ and stir 0.5～1.5h; Then add glutaraldehyde, regulate pH to 8～12, then be warming up to 30～80 ℃, continue reaction 2～6h, suction filtration, washing, drying obtain the shitosan skeleton while hot; The volume ratio that feeds intake of described chitosan solution and atoleine is 1: 0.5～5, be preferably 1: 1～and 3; The mol ratio of described crosslinking agent glutaraldehyde and amino of chitosan is 1: 0.1～10, be preferably 1: 0.5～and 3; The volume ratio that feeds intake of described perforating agent cyclohexane and atoleine is 5～50: 100, is preferably 10～30: 100.The consumption of described span 80 only needs a little to get final product, and as in the 100mL atoleine, only need add 5-10 to drip span 80 and get final product.
Further, in step (1), preferably be heated to 50～60 ℃ and stir 0.5～1.0h; Then add glutaraldehyde, regulate pH to 9～10, then be warming up to 60～70 ℃, continue reaction 2.5～4h.
Step of the present invention (2) activates the shitosan skeleton that step (1) makes, and introduces epoxy radicals on the shitosan skeleton.Further, the present invention specifically recommends described step (2) to carry out according to following: take the shitosan skeleton, after the abundant swelling of water, the dimethyl sulphoxide solution with 10%～100% gradient concentration cleans successively; Add DMSO/NaOH solution and epoxychloropropane in product after process, at 20～80 ℃ of oscillating reactions 1～10h, clean with distilled water after reaction finishes, until detect without epoxy radicals in cleaning fluid; Described DMSO/NaOH mixed liquor is obtained according to volume ratio 1: 0.1～10.0 preparations by the NaOH aqueous solution of DMSO and 0.1～1.0mol/L, preferably obtains according to volume ratio 1: 0.5～2.0 preparations; The volume ratio that feeds intake of described epoxychloropropane and DMSO/NaOH mixed liquor is 1～20: 100, is preferably 5～15: 100.
Further, in step (2), preferably 40～50 ℃ of oscillating reactions, preferred 3～5h of reaction time.
In said method, shitosan skeleton pressed on ring oxygen base density can be measured by thio sulfate method.
In step of the present invention (3), in described IDA/NaOH mixed solution, the preferred 1mol/L of IDA concentration, the preferred 1mol/L of NaOH concentration.Preferred 55～65 ℃ of the reaction temperature of step (3), preferred 3～8h of reaction time.
Compared with prior art, beneficial effect of the present invention is: shitosan separating medium preparation method of the present invention is simple to operate, and granularity uniformity obviously improves, has very strong acid-fast alkali-proof performance, and has more amino, metal ion is had stronger sequestering power, and epoxy group modified density reachable is to 0.1mmol/g, Cu
2+The chelating amount obviously improves, and has solved to a great extent the problem that metal ion is revealed, and the BSA maximal absorptive capacity can reach 128mg/g, has technology stability and high repeatability and other advantages, suitability for scale production.
Four. description of drawings
Fig. 1 is the schematic arrangement of the shitosan separating medium for preparing of the present invention.
Fig. 2 is the scanning electron microscope (SEM) photograph of the shitosan separating medium for preparing of the present invention.
Fig. 3 is the particle diameter distribution map of the shitosan separating medium for preparing of the present invention.
Five. the specific embodiment
The below is described further technical scheme of the present invention with specific embodiment, but protection scope of the present invention is not limited to this:
The preparation of embodiment 1 shitosan skeleton
It is in 2.0% acetum that 3.0g shitosan (molecular weight is 30000Da, deacetylation 〉=95%) is dissolved in the 100mL mass fraction, and hold over night under room temperature is standby.In the 500mL there-necked flask that mechanical agitator and thermometer are housed, add successively atoleine 100mL, cyclohexane 15mL and 6 Span 80, after stirring 0.5h, add above-mentioned chitosan solution, with water-bath, system is heated to 55 ℃, stirs 1h, adding mass fraction is 25% glutaraldehyde 3mL; With 10%NaOH solution adjust pH to 10, then be warming up to 65 ℃, after continuing reaction 3h, with the vacuum filtration pump, the microballoon that obtains is leached while hot, after repeatedly washing with distilled water, then use benzinum and absolute ethanol washing, vacuum drying is to constant weight, make the shitosan skeleton, 90% particle size distribution is at 125～200 μ m.
Embodiment 2 chitosan grafts
Take the product that 0.5g embodiment 1 obtains, after the abundant swelling of water, use successively 20%, 50%, 70% DMSO aqueous cleaning; Add 27mL DMSO/NaOH solution (the DMSO volume fraction is that 0.4, NaOH concentration of aqueous solution is 0.4mol/L) and epoxychloropropane in product after process, the volume fraction that makes epoxychloropropane is 10%, oscillating reactions 4h, and reaction temperature is 50 ℃.Reaction is used a large amount of distilled water flushings after finishing, until detect without epoxy radicals in cleaning fluid.The epoxy group modified density of shitosan adopts sodium thiosulfate titration, and its epoxy group modified density reaches 0.1mmol/g.
Embodiment 3 chitosan grafts
Take the product that 0.5g embodiment 1 obtains, after the abundant swelling of water, use successively 20%, 50%, 70% DMSO aqueous cleaning; Add 27mL DMSO/NaOH solution (the DMSO volume fraction is that 0.5, NaOH concentration of aqueous solution is 0.4mol/L) and epoxychloropropane in product after process, the volume fraction that makes epoxychloropropane is 6%, oscillating reactions 3h, and reaction temperature is 40 ℃.Reaction is used a large amount of distilled water flushings after finishing, until detect without epoxy radicals in cleaning fluid.The epoxy group modified density of shitosan adopts sodium thiosulfate titration, and its epoxy group modified density reaches 0.089mmol/g.
The preparation of embodiment 4 shitosan separating mediums
50mL IDA/NaOH mixed solution (concentration of IDA and NaOH is respectively 1.0mol/L) is added microballoon after the grafting that embodiment 2 makes, react 3h at 60 ℃ of temperature, the product suction filtration that obtains is got final product.
Cu
2+The mensuration of chelating amount: accurately take a certain amount of shitosan separating medium that makes, add the CuSO with the same concentration of volume (0.1mol/L)
4Chelating Cu in solution
2+, after vacuum filtration, measure residual Cu in filtrate
2+The i.e. Cu as can be known of amount
2+The chelating amount.AAS (wavelength is 660nm) is measured copper ion content of solution, Cu
2+The chelating amount reaches 162.0mg/g.
The mensuration of bovine serum albumin(BSA) (BSA) adsorption capacity: get the shitosan separating medium 1mL that makes, BSA upper prop with 5mL, 10mg/mL, flow velocity 20mL/h, Tris-HCl buffer solution elution with 0.05mol/L, pH7.5, eluent is collected in on-line monitoring, measure albumen in eluent, get the BSA adsorption capacity.The mensuration of BSA concentration adopts Coomassie brilliant blue method (595nm).The maximal absorptive capacity of BSA reaches 128.0mg/g.
The preparation of embodiment 5 shitosan separating mediums
50mL IDA/NaOH mixed solution (concentration of IDA and NaOH is respectively 1.0mol/L) is added microballoon after the grafting that embodiment 3 makes, react 3h at 60 ℃ of temperature, the product suction filtration that obtains is got final product.
Cu
2+The assay method of chelating amount and BSA adsorbance is with embodiment 4, and result shows: adopt AAS (wavelength is 660nm) to measure Cu
2+The chelating amount, its chelating amount reaches 103.8mg/g; Utilize Coomassie brilliant blue method (595nm) to measure BSA content, its maximal absorptive capacity reaches 89.2mg/g.
Claims (8)
1. a preparation method who is suitable for the shitosan separating medium of protein purification, comprise the steps:
(1) under agitation, the acetic acid solution of shitosan is scattered in atoleine, under perforating agent cyclohexane and a little span80 exist, forms chitosan particle; Then under the effect of crosslinking agent glutaraldehyde, chitosan particle further is cross-linked to form the shitosan skeleton; The volume ratio that feeds intake of described chitosan solution and atoleine is 1 ︰ 0.5 ~ 5, and the mol ratio of described crosslinking agent glutaraldehyde and amino of chitosan is 1 ︰ 0.1 ~ 10, and described perforating agent cyclohexane and the atoleine volume ratio that feeds intake is 5 ~ 50 ︰ 100;
(2) the advanced row of shitosan skeleton swelling treatment; Then in the DMSO/NaOH mixed liquor, the hydroxyl on the shitosan skeleton and epichlorohydrin reaction are introduced epoxy radicals on the shitosan skeleton, obtain the shitosan skeleton after grafting; Described DMSO/NaOH mixed liquor is obtained according to volume ratio 1 ︰ 0.1～10.0 preparation by the NaOH aqueous solution of DMSO and 0.1～1.0mol/L; The volume ratio that feeds intake of described epoxychloropropane and DMSO/NaOH mixed liquor is 1～20 ︰ 100;
(3) the IDA/NaOH mixed solution is added shitosan skeleton after grafting, react 1 ~ 10h at 20 ~ 80 ℃ of temperature, suction filtration namely gets described shitosan separating medium; In described IDA/NaOH mixed solution, IDA concentration is 0.5 ~ 3mol/L, and NaOH concentration is 0.5 ~ 3.0mol/L.
2. the preparation method who is suitable for the shitosan separating medium of protein purification as claimed in claim 1 is characterized in that described step (1) is specific as follows: it is that in 1.0 ~ 3.0% acetum, preparation obtains the chitosan solution that concentration is 0.01 ~ 0.05g/mL that shitosan is dissolved in mass fraction; Add successively atoleine, cyclohexane and a little Span80 in reaction vessel, stir the chitosan solution that adds preparation to obtain after 30 ~ 60min, be heated to 20 ~ 80 ℃ and stir 0.5 ~ 1.5h; Then add glutaraldehyde, regulate pH to 8 ~ 12, then be warming up to 30 ~ 80 ℃, continue reaction 2 ~ 6h, suction filtration, washing, drying obtain the shitosan skeleton while hot; The volume ratio that feeds intake of described chitosan solution and atoleine is 1 ︰ 0.5 ~ 5, and the mol ratio of described crosslinking agent glutaraldehyde and amino of chitosan is 1 ︰ 0.1 ~ 10, and described perforating agent cyclohexane and the atoleine volume ratio that feeds intake is 5 ~ 50 ︰ 100.
3. the preparation method who is suitable for the shitosan separating medium of protein purification as claimed in claim 2, is characterized in that in described step (1), and the volume ratio that feeds intake of described chitosan solution and atoleine is 1 ︰ 1 ~ 3; The mol ratio of described crosslinking agent glutaraldehyde and amino of chitosan is 1 ︰ 0.5 ~ 3; The volume ratio that feeds intake of described perforating agent cyclohexane and atoleine is 10 ~ 30 ︰ 100.
4. the preparation method who is suitable for the shitosan separating medium of protein purification as claimed in claim 2, is characterized in that in described step (1), first is heated to 50 ~ 60 ℃ after the chitosan solution that adds preparation to obtain and stirs 0.5 ~ 1.0h; Then add glutaraldehyde, regulate pH to 9 ~ 10, then be warming up to 60 ~ 70 ℃, continue reaction 2.5 ~ 4h.
5. the preparation method who is suitable for the shitosan separating medium of protein purification as claimed in claim 1, it is characterized in that described step (2) is specific as follows: take the shitosan skeleton, after the abundant swelling of water, the dimethyl sulphoxide solution that is 10%～100% gradient concentration with volume fraction successively cleans; Add DMSO/NaOH solution and epoxychloropropane in product after process, at 20 ~ 80 ℃ of oscillating reactions 1 ~ 10h, clean with distilled water after reaction finishes, until detect without epoxy radicals in cleaning fluid.
6. the preparation method who is suitable for the shitosan separating medium of protein purification as claimed in claim 5, it is characterized in that in described step (2), described DMSO/NaOH mixed liquor is obtained according to volume ratio 1 ︰ 0.5～2.0 preparation by the NaOH aqueous solution of DMSO and 0.1～1.0mol/L; The volume ratio that feeds intake of described epoxychloropropane and DMSO/NaOH mixed liquor is 5～15 ︰ 100.
7. the preparation method who is suitable for the shitosan separating medium of protein purification as claimed in claim 5, is characterized in that in described step (2), at 40 ~ 50 ℃ of oscillating reactions, reaction time 3 ~ 5h.
8. the preparation method who is suitable for the shitosan separating medium of protein purification as claimed in claim 1, is characterized in that in described step (3), reaction temperature is 55 ~ 65 ℃, and the reaction time is 3 ~ 8h.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010579423 CN102068965B (en) | 2010-12-09 | 2010-12-09 | Method for preparing chitosan separation medium suitable for protein purification |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201010579423 CN102068965B (en) | 2010-12-09 | 2010-12-09 | Method for preparing chitosan separation medium suitable for protein purification |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102068965A CN102068965A (en) | 2011-05-25 |
| CN102068965B true CN102068965B (en) | 2013-06-05 |
Family
ID=44027845
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 201010579423 Active CN102068965B (en) | 2010-12-09 | 2010-12-09 | Method for preparing chitosan separation medium suitable for protein purification |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102068965B (en) |
Families Citing this family (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104277111B (en) * | 2013-07-08 | 2020-06-12 | 百瑞全球有限公司 | Composite carrier for preparing immobilized protein, polypeptide or oligopeptide, preparation method and application |
| CN104861181B (en) * | 2015-04-15 | 2017-08-22 | 中华人民共和国广州机场出入境检验检疫局 | A kind of preparation method of the chitosan microball of high intensity |
| CN105131329B (en) * | 2015-10-16 | 2018-03-30 | 武汉科技大学 | A kind of preparation method and application of the polyvinyl alcohol crosslinked affinity membrane of macropore chitosan of chelated metal ions |
| CN105602717A (en) * | 2015-12-28 | 2016-05-25 | 山东润泽制药有限公司 | Refining method of horse oil |
| CN106423079A (en) * | 2016-10-17 | 2017-02-22 | 江苏大学 | Preparation method and application of chitosan-microsphere immunoaffinity adsorbent |
| CN106754861B (en) * | 2016-12-26 | 2020-09-25 | 浙江工商大学 | Porous magnetic copper ion metal chelating carrier and preparation method thereof, and method for immobilizing papain by using carrier and application thereof |
| CN106676093B (en) * | 2016-12-26 | 2020-01-07 | 浙江工商大学 | Porous magnetic nickel ion metal chelating carrier and preparation method thereof, and method for immobilizing papain by using carrier and application thereof |
| CN107226900B (en) * | 2017-06-20 | 2019-03-15 | 吉林省爱诺德生物工程有限公司 | A kind of aflatoxins B1The preparation method of molecularly imprinted polymer |
| CN107163226B (en) * | 2017-06-20 | 2019-01-15 | 吉林省爱诺德生物工程有限公司 | A kind of preparation method of Ochratoxin A molecularly imprinted polymer |
| CN108956488B (en) * | 2018-04-23 | 2020-11-10 | 山东省医疗器械产品质量检验中心 | Method for measuring protein content in chitosan or chitosan salt |
| CN113231049B (en) * | 2021-05-11 | 2022-11-01 | 南京工业大学 | Cross-linked agarose affinity medium, and preparation method and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1359750A (en) * | 2001-09-25 | 2002-07-24 | 广东梅县梅雁蓝藻有限公司 | Process for synthesizing heavy metal adsorbent from silica gel and chitosan |
| CN101037486A (en) * | 2007-04-27 | 2007-09-19 | 中国林业科学研究院林产化学工业研究所 | Preparation method of quaternary ammonium N,O-carboxyetbyl chitosan |
| CN101085359A (en) * | 2007-06-14 | 2007-12-12 | 上海师范大学 | Magnetic chitosan medicine-carried nano particles and preparation method thereof |
| CN101747448A (en) * | 2008-11-28 | 2010-06-23 | 北京大学 | Nano chitosan derivative and preparation method and application thereof |
-
2010
- 2010-12-09 CN CN 201010579423 patent/CN102068965B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1359750A (en) * | 2001-09-25 | 2002-07-24 | 广东梅县梅雁蓝藻有限公司 | Process for synthesizing heavy metal adsorbent from silica gel and chitosan |
| CN101037486A (en) * | 2007-04-27 | 2007-09-19 | 中国林业科学研究院林产化学工业研究所 | Preparation method of quaternary ammonium N,O-carboxyetbyl chitosan |
| CN101085359A (en) * | 2007-06-14 | 2007-12-12 | 上海师范大学 | Magnetic chitosan medicine-carried nano particles and preparation method thereof |
| CN101747448A (en) * | 2008-11-28 | 2010-06-23 | 北京大学 | Nano chitosan derivative and preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 王晓军等.壳聚糖金属螯合亲和吸附剂的制备.《西北大学学报（自然科学版）》.2009,第39卷(第4期),第595-598，602页. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102068965A (en) | 2011-05-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102068965B (en) | Method for preparing chitosan separation medium suitable for protein purification | |
| CN102732475B (en) | A kind of microcarrier for cell cultures, its preparation method and detection method | |
| KR102624853B1 (en) | Endotoxin adsorbent | |
| EP1924345A2 (en) | Process for cross-linking cellulose ester membranes | |
| JP2002263486A (en) | Endotoxin adsorbent and method of removing endotoxin by using the same | |
| CN107893062B (en) | Method for immobilizing cellulase and hydrolyzing cellulose | |
| Santos-Moriano et al. | Vinyl sulfone-activated silica for efficient covalent immobilization of alkaline unstable enzymes: Application to levansucrase for fructooligosaccharide synthesis | |
| WO2022088220A1 (en) | Pmma matrix-based protein a affinity chromatography medium and preparation method and application thereof | |
| CN106622401B (en) | A kind of preparation method of the high carrying capacity ionic energy transfer purifying micro-sphere material of hydrophilic | |
| EP2266675A2 (en) | Chromatography medium, preparation method of the same, and method for producing virus vaccine using the chromatography medium | |
| CN101716494B (en) | Magnetic compatible microsphere for purifying thrombin and preparation method and application thereof | |
| CN110152624A (en) | A kind of hydrophilic resin that microporous polymer coats and its application in glycopeptide enrichment | |
| CN102019213B (en) | Method for preparing strongly-acid ion exchange medium | |
| CN109158089A (en) | A kind of sulfhydryl modified cellulose aerogels of ultrasonic wave added and preparation method | |
| CN105363417A (en) | Preparation method for cross-linked carboxymethylated agarose-base gel microsphere | |
| CN102559648A (en) | Immobilized enzyme using modified epoxy resin as carrier and preparation method and application thereof | |
| Valentova et al. | Comparison of different methods of glucose oxidase immobilization | |
| CN106632529B (en) | A kind of shell tetrose monomer separation extracting method based on molecular imprinting technology | |
| CN108864364B (en) | Preparation method of L-phenylalanine molecularly imprinted polymer | |
| CN107096509B (en) | It is a kind of containing α-amido succinic acid function base sephadex and preparation method | |
| CN104694454B (en) | A kind of microcarrier for cell culture and its preparation method and application | |
| Jirku et al. | [30] Cell immobilization by covalent linkage | |
| CN103965484B (en) | Preparation method and application of omega-diamine derivatization beta-cyclodextrin bonded SBA-15 chiral stationary phase | |
| CN113117657B (en) | Preparation and application of beta-cyclodextrin metal organic framework material HPLC column | |
| CN101284224B (en) | Expanded bed adsorption medium with sulfhydryl ethylpyridine and sulfone group as ligand separation antibody and preparation method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |