CN105837748B - A kind of stratum's ordered big hole mesoporous silica gel integral post bovine serum albumin(BSA) imprinted polymer and preparation method thereof - Google Patents
A kind of stratum's ordered big hole mesoporous silica gel integral post bovine serum albumin(BSA) imprinted polymer and preparation method thereof Download PDFInfo
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- CN105837748B CN105837748B CN201610192313.0A CN201610192313A CN105837748B CN 105837748 B CN105837748 B CN 105837748B CN 201610192313 A CN201610192313 A CN 201610192313A CN 105837748 B CN105837748 B CN 105837748B
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- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 title claims abstract description 62
- 108091003079 Bovine Serum Albumin Proteins 0.000 title claims abstract description 43
- 229940098773 bovine serum albumin Drugs 0.000 title claims abstract description 43
- 239000000741 silica gel Substances 0.000 title claims abstract description 40
- 229910002027 silica gel Inorganic materials 0.000 title claims abstract description 40
- 229920000642 polymer Polymers 0.000 title claims abstract description 32
- 238000002360 preparation method Methods 0.000 title claims abstract description 13
- 239000004793 Polystyrene Substances 0.000 claims abstract description 30
- 229920002223 polystyrene Polymers 0.000 claims abstract description 29
- 239000000084 colloidal system Substances 0.000 claims abstract description 21
- 239000013078 crystal Substances 0.000 claims abstract description 21
- 230000015572 biosynthetic process Effects 0.000 claims abstract description 16
- 238000003786 synthesis reaction Methods 0.000 claims abstract description 16
- 239000000463 material Substances 0.000 claims abstract description 7
- 238000001179 sorption measurement Methods 0.000 claims abstract description 4
- -1 γ (methacryloxypropyl) propyl Chemical group 0.000 claims abstract description 4
- BLRPTPMANUNPDV-UHFFFAOYSA-N Silane Chemical compound [SiH4] BLRPTPMANUNPDV-UHFFFAOYSA-N 0.000 claims abstract 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 29
- 239000000243 solution Substances 0.000 claims description 28
- ROOXNKNUYICQNP-UHFFFAOYSA-N ammonium peroxydisulfate Substances [NH4+].[NH4+].[O-]S(=O)(=O)OOS([O-])(=O)=O ROOXNKNUYICQNP-UHFFFAOYSA-N 0.000 claims description 25
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 24
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 24
- 238000006243 chemical reaction Methods 0.000 claims description 20
- 239000007788 liquid Substances 0.000 claims description 18
- 229910001870 ammonium persulfate Inorganic materials 0.000 claims description 14
- 238000001291 vacuum drying Methods 0.000 claims description 14
- HRPVXLWXLXDGHG-UHFFFAOYSA-N Acrylamide Chemical compound NC(=O)C=C HRPVXLWXLXDGHG-UHFFFAOYSA-N 0.000 claims description 13
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 13
- LYCAIKOWRPUZTN-UHFFFAOYSA-N ethylene glycol Natural products OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims description 10
- WGCNASOHLSPBMP-UHFFFAOYSA-N hydroxyacetaldehyde Natural products OCC=O WGCNASOHLSPBMP-UHFFFAOYSA-N 0.000 claims description 10
- BOTDANWDWHJENH-UHFFFAOYSA-N Tetraethyl orthosilicate Chemical compound CCO[Si](OCC)(OCC)OCC BOTDANWDWHJENH-UHFFFAOYSA-N 0.000 claims description 9
- 239000008367 deionised water Substances 0.000 claims description 9
- 229910021641 deionized water Inorganic materials 0.000 claims description 9
- 239000000377 silicon dioxide Substances 0.000 claims description 8
- KWYHDKDOAIKMQN-UHFFFAOYSA-N N,N,N',N'-tetramethylethylenediamine Chemical compound CN(C)CCN(C)C KWYHDKDOAIKMQN-UHFFFAOYSA-N 0.000 claims description 7
- RCEAADKTGXTDOA-UHFFFAOYSA-N OS(O)(=O)=O.CCCCCCCCCCCC[Na] Chemical compound OS(O)(=O)=O.CCCCCCCCCCCC[Na] RCEAADKTGXTDOA-UHFFFAOYSA-N 0.000 claims description 7
- 235000019441 ethanol Nutrition 0.000 claims description 6
- 239000013590 bulk material Substances 0.000 claims description 5
- 238000007598 dipping method Methods 0.000 claims description 5
- 125000005909 ethyl alcohol group Chemical group 0.000 claims description 5
- 238000012986 modification Methods 0.000 claims description 5
- 230000004048 modification Effects 0.000 claims description 5
- 239000008363 phosphate buffer Substances 0.000 claims description 5
- 238000006116 polymerization reaction Methods 0.000 claims description 5
- 239000011148 porous material Substances 0.000 claims description 5
- 238000002604 ultrasonography Methods 0.000 claims description 5
- 239000007864 aqueous solution Substances 0.000 claims description 4
- 238000001704 evaporation Methods 0.000 claims description 4
- 230000008020 evaporation Effects 0.000 claims description 4
- 239000000047 product Substances 0.000 claims description 4
- ULWHHBHJGPPBCO-UHFFFAOYSA-N propane-1,1-diol Chemical class CCC(O)O ULWHHBHJGPPBCO-UHFFFAOYSA-N 0.000 claims description 4
- ZIUHHBKFKCYYJD-UHFFFAOYSA-N n,n'-methylenebisacrylamide Chemical compound C=CC(=O)NCNC(=O)C=C ZIUHHBKFKCYYJD-UHFFFAOYSA-N 0.000 claims description 3
- 102000009027 Albumins Human genes 0.000 claims description 2
- 108010088751 Albumins Proteins 0.000 claims description 2
- 229910019142 PO4 Inorganic materials 0.000 claims description 2
- 230000003139 buffering effect Effects 0.000 claims description 2
- 239000011259 mixed solution Substances 0.000 claims description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 2
- 239000010452 phosphate Substances 0.000 claims description 2
- 210000002966 serum Anatomy 0.000 claims description 2
- 239000006228 supernatant Substances 0.000 claims description 2
- VAZSKTXWXKYQJF-UHFFFAOYSA-N ammonium persulfate Chemical compound [NH4+].[NH4+].[O-]S(=O)OOS([O-])=O VAZSKTXWXKYQJF-UHFFFAOYSA-N 0.000 claims 2
- YUYCVXFAYWRXLS-UHFFFAOYSA-N trimethoxysilane Chemical compound CO[SiH](OC)OC YUYCVXFAYWRXLS-UHFFFAOYSA-N 0.000 claims 2
- 241000283690 Bos taurus Species 0.000 claims 1
- 238000002242 deionisation method Methods 0.000 claims 1
- 230000035699 permeability Effects 0.000 abstract description 3
- 230000001105 regulatory effect Effects 0.000 abstract description 3
- 238000005516 engineering process Methods 0.000 description 6
- 102000004169 proteins and genes Human genes 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- PMZURENOXWZQFD-UHFFFAOYSA-L Sodium Sulfate Chemical compound [Na+].[Na+].[O-]S([O-])(=O)=O PMZURENOXWZQFD-UHFFFAOYSA-L 0.000 description 5
- 229910052938 sodium sulfate Inorganic materials 0.000 description 5
- 235000011152 sodium sulphate Nutrition 0.000 description 5
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical group [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 4
- 229920000344 molecularly imprinted polymer Polymers 0.000 description 4
- 239000002243 precursor Substances 0.000 description 4
- 238000000926 separation method Methods 0.000 description 4
- 238000002156 mixing Methods 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 230000005540 biological transmission Effects 0.000 description 2
- 238000009826 distribution Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000002329 infrared spectrum Methods 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 239000013335 mesoporous material Substances 0.000 description 2
- 239000003960 organic solvent Substances 0.000 description 2
- 239000002245 particle Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 150000005846 sugar alcohols Polymers 0.000 description 2
- 238000007445 Chromatographic isolation Methods 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
- 230000002902 bimodal effect Effects 0.000 description 1
- 238000001354 calcination Methods 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000008602 contraction Effects 0.000 description 1
- 239000003431 cross linking reagent Substances 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 150000004985 diamines Chemical class 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 150000004678 hydrides Chemical class 0.000 description 1
- 239000003999 initiator Substances 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 238000002414 normal-phase solid-phase extraction Methods 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08F—MACROMOLECULAR COMPOUNDS OBTAINED BY REACTIONS ONLY INVOLVING CARBON-TO-CARBON UNSATURATED BONDS
- C08F222/00—Copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by a carboxyl radical and containing at least one other carboxyl radical in the molecule; Salts, anhydrides, esters, amides, imides, or nitriles thereof
- C08F222/36—Amides or imides
- C08F222/38—Amides
- C08F222/385—Monomers containing two or more (meth)acrylamide groups, e.g. N,N'-methylenebisacrylamide
-
- 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
- B01J20/268—Polymers created by use of a template, e.g. molecularly imprinted polymers
-
- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J9/00—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof
- C08J9/26—Working-up of macromolecular substances to porous or cellular articles or materials; After-treatment thereof by elimination of a solid phase from a macromolecular composition or article, e.g. leaching out
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2201/00—Foams characterised by the foaming process
- C08J2201/04—Foams characterised by the foaming process characterised by the elimination of a liquid or solid component, e.g. precipitation, leaching out, evaporation
- C08J2201/042—Elimination of an organic solid phase
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08J—WORKING-UP; GENERAL PROCESSES OF COMPOUNDING; AFTER-TREATMENT NOT COVERED BY SUBCLASSES C08B, C08C, C08F, C08G or C08H
- C08J2335/00—Characterised by the use of homopolymers or copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by a carboxyl radical, and containing at least one other carboxyl radical in the molecule, or of salts, anhydrides, esters, amides, imides or nitriles thereof; Derivatives of such polymers
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Analytical Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Materials Engineering (AREA)
- Solid-Sorbent Or Filter-Aiding Compositions (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention provides a kind of stratum's ordered big hole mesoporous silica gel integral post bovine serum albumin(BSA) imprinted polymer and preparation method thereof, including：The synthesis of polystyrene colloid crystal；The synthesis of stratum's ordered big hole mesoporous silica gel integral post；Prepare stratum's ordered big hole mesoporous silica gel integral post γ (methacryloxypropyl) propyl trimethoxy silicane decorative material；The preparation of stratum's ordered big hole mesoporous silica gel integral post bovine serum albumin(BSA) imprinted polymer.The synthesized well-regulated macroscopic view of stratum's ordered big hole mesoporous silica gel integral post tool and microstructure, permeability is strong, and back pressure is low, and mass transfer dynamicses are fast.The bovine serum albumin(BSA) imprinted polymer of synthesis, recognition site are located near or at polymer surfaces, facilitate bovine serum albumin(BSA) molecule to pass in and out imprinted cavity, improve the selectivity to bovine serum albumin(BSA).Stratum's ordered big hole mesoporous silica gel integral post bovine serum albumin(BSA) imprinted polymer has good separating property to bovine serum albumin(BSA), and adsorption capacity is up to 121.6mg/g.
Description
Technical field
The present invention relates to monolithic silica column imprinted polymer, more particularly to a kind of ordered porous monolithic silica column trace of stratum
Polymer and preparation method thereof, belong to material preparation and technical field of bioengineering.
Background technology
SPE (Solid-Phase Extraction, SPE) is that developed recently gets up a kind of Sample Pretreatment Technique Used,
It is mainly used in the isolation and purification of sample.It is simple to operate, time saving and energy saving, can be more effectively by analyte and interfering component point
From being widely used in the fields such as medicine, food, chemical industry.The splitter of SPE is generally by the particles filled of controlled dimensions.
Grain diameter is smaller, and the efficiency of separation is higher, but back pressure also rises simultaneously.Recently, the concept of integral post is introduced into isolation technics.
Overall rod structure can eliminate the space of particle intercolumniation, so as to cause in separation process faster mass transfer dynamicses.It is in addition, whole
The permeability enhancing of scapus, back pressure is extremely low, reduces the obstruction of post bed, can choose the separation property that higher bed reaches higher
Energy.
Integral post can be divided into two classes：Polyalcohol integral pole and monolithic silica column.Polyalcohol integral pole has under different pH
Good stability, and there is good biocompatibility.But it can contraction or expansion under the influence of temperature and organic solvent.
Monolithic silica column is the porous rod being made up of the silicon skeleton of interconnection macropore, there is a large amount of mesoporous presence on skeleton.It is whole with polymer
Scapus is compared, and monolithic silica column organic solvent-resistant, mechanical strength is stronger, and post rate is higher.But monolithic silica column mesopore is generally mixed and disorderly
Unordered, pore-size distribution is wide, causes the inhomogeneity of mass transfer, influences post effect.
In recent years, by nature generally existing the orderly pore structure of stratum (such as：Diatom, lumbar vertebrae, lung) inspiration, largely
Research worker synthesis with different pore size stratum's Ordered Materials.Stratum's ordered big hole-mesoporous material, attractive rule
Macroporous structure then, passage can be served as and facilitate mass transport；Simultaneously there is regular meso-hole structure, there is provided high-specific surface area and
High pore volume.Therefore, stratum's ordered big hole-mesoporous material can make target molecule more easily be diffused into whole bimodal hole system
System, and easily accessible binding site is provided, catalysis, absorption and biotechnology field can be widely used in.
Molecular imprinting technology is technology emerging in recent years, and it creates high with template molecule shape, size and space structure
Consistent adsorption site is spent, the molecularly imprinted polymer (MIP) prepared has excellent mechanical/chemical stability, low cost
And the high specific to target molecule, it is widely used in the fields such as chromatographic isolation, SPE, sensor.Up to the present, permitted
More small molecule imprinted polymers are successfully prepared, and have excellent performance.However, protein-imprinted polymer few reports
Road, especially macro-molecular protein imprinted polymer.The molecular dimension of protein is larger and with complicated conformation, mass transfer
Difficult and shortage selectivity.In order to solve this problem, surface imprinted technology is arisen at the historic moment.During surface imprinted, trace position
Point is located near or at MIP surfaces, is easy to target molecule to pass in and out imprinted cavity.
Bovine serum albumin(BSA) (BSA) is one of seralbumin, molecular mass 66.5kDa, in biologic applications and enzymatic
Reaction field is widely used.It is the protein reagent commonly used in protein determination, measures a kind of concentration of protein in solution.This
Outside, bovine serum albumin(BSA) is similar to human serum albumins (HSA) structure, and BSA passes through the model protein frequently as research HSA.So
It is most important to BSA Selective Separation.
The content of the invention
The purpose of the present invention is with polystyrene (PS) colloidal crystal and poly- (1,2- ethylidene glycol) block poly- (the third two
Alcohol) block poly- (1,2- ethylidene glycol) (P123) is used as double template, synthesis stratum ordered big hole-mesoporous silica gel integral post
(HOM).Then using HOM as host material, bovine serum albumin(BSA) molecularly imprinted polymer is synthesized using surface imprinted technology.Synthesis
Stratum's ordered big hole-mesoporous silica gel integral post imprinted polymer there is good macro-and micro-structure, have to BSA good
Separating effect.
The present invention is achieved through the following technical solutions：
A kind of stratum's ordered big hole-mesoporous silica gel integral post bovine serum albumin(BSA) imprinted polymer, stratum's ordered big hole-
Mesoporous silica gel integral post bovine serum albumin(BSA) imprinted polymer is made up of silica skeleton and macropore, is had on silica skeleton
Even distribution it is mesoporous；The macropore diameter is 1~1.2 μm, the structure interconnected is formed between macropore, the mesoporous pore size is
6~7nm；The imprinted polymer is bovine serum albumin(BSA), and the bovine serum albumin(BSA) is uniformly modified on silica skeleton surface.
A kind of preparation method of stratum's ordered big hole-mesoporous silica gel integral post bovine serum albumin(BSA) imprinted polymer, including
Following steps：
The synthesis of step 1, polystyrene colloid crystal：Polystyrene is dissolved in a certain amount of water, polystyrene and water
Amount ratio is 0.4~1g：8mL.By solution ultrasound 20min, then natural subsidence one week, carefully removes supernatant liquid, and 25 DEG C true
Sky dries 48h, obtains polystyrene colloid crystal；
The synthesis of step 2, stratum's ordered big hole-mesoporous silica gel integral post (HOM)：A certain amount of tetraethyl orthosilicate
(TEOS), water, hydrochloric acid, poly- poly- (propane diols) block of (1,2- ethylidene glycol) block poly- (1,2- ethylidene glycol) (P123) and
Absolute ethyl alcohol, which is prepared, turns into precursor solution, and polystyrene colloid crystal is added in solution, impregnates 1h, obtains mixed liquor A,
Wherein concentration of hydrochloric acid is 2mol/L；Then by mixed liquor A in 30 DEG C of dipping evaporation 48h；The bulk material of synthesis is placed in tubular type
In stove, in atmosphere with 1 DEG C of min-1Programming rate slowly heat up, to constant temperature 6h at 550 DEG C, obtain stratum's ordered big hole-
Mesoporous silica gel integral post (HOM)；
Step 3, prepare stratum's ordered big hole-mesoporous silica gel integral post γ-(methacryloxypropyl) propyl trimethoxy
Hydride modified material, referred to as HOM@MPS：The HOM of gained is dissolved in 40mL absolute ethyl alcohols in 0.4g steps 2, then takes 2mL
γ-(methacryloxypropyl) propyl trimethoxy silicane (MPS) is added in mixed solution, 40 DEG C of reaction 12h；Will production after reaction
Thing is washed 3 times with absolute ethyl alcohol, and to elute unreacted MPS, 50 DEG C of vacuum drying obtain the HOM, referred to as HOM@of MPS modifications
MPS；
The system of step 4, stratum's ordered big hole-mesoporous silica gel integral post bovine serum albumin(BSA) imprinted polymer (BSA-MIP)
It is standby：The HOM@MPS that step 3 obtains are scattered in phosphate buffer, ultrasonic 10min, mixed liquid B are obtained, by acrylamide
(AM), N, N '-methylene-bisacrylamide (MBA), bovine serum albumin(BSA) (BSA) is added in mixed liquid B, obtains mixed liquor C,
1h is stirred at 35 DEG C to mixed liquor C, leads to N2, ammonium persulfate (APS) and N, N, N, N- tetramethylethylenediamine (TEMED) are added to
In solution, 35 DEG C of reaction 24h；After reaction, 5 times are washed repeatedly with deionized water to remove unreacted AM, APS and BSA, 25
DEG C vacuum drying；Then with the acetic acid of lauryl sodium sulfate (SDS) of the 500~1000mL containing 100mg/L：The aqueous solution washes away
BSA template molecules, and being washed with deionized water to neutrality, 25 DEG C are dried in vacuo and produce stratum's ordered big hole-mesoporous silica gel integral post
Bovine serum albumin(BSA) imprinted polymer (BSA-MIP).
In step 2, when preparing mixed liquor A, tetraethyl orthosilicate used, water, hydrochloric acid, poly- (1,2- ethylidene glycol) is embedding
Poly- (propane diols) block of section is poly- (1,2- ethylidene glycol), the mass ratio of absolute ethyl alcohol and polystyrene colloid crystal for 1.5~
2.5g：0.9g：0.1~0.8g：1.0g：15g：0.4~1g.
In step 4, when preparing mixed liquid B, HOM@MPS concentration is 2.5~5mg/mL；It is used when preparing mixed liquor C
HOM@MPS, acrylamide, N, N '-methylene-bisacrylamide, bovine serum albumin(BSA), ammonium persulfate, N, N, N, N- tetramethyl second
The amount ratio of diamines is 0.05~0.10g：35.5~71.0mg：77mg：10~20mg：8~10mg：10μL.
In step 4, the acetic acid of the lauryl sodium sulfate (SDS) containing 100mg/L：In the aqueous solution, acetic acid and water
Volume ratio be 1：9.
In step 2, described TEOS, it act as silicon source.
In step 2, described P123, it act as mesoporous template.
In step 2, described polystyrene colloid crystal, it act as macropore template.
In step 4, described phosphate buffering liquid concentration is 20mmol/L, and pH is 7.0~8.0.
In step 4, described HOM@MPS, it act as host material.
In step 4, described AM, it act as function monomer.
In step 4, described MBA, it act as crosslinking agent.
In step 4, described BSA, it act as template molecule.
In step 4, described APS and TEMED, it act as initiator.
The technological merit of the present invention：
(1) the well-regulated macroscopic view of stratum's ordered big hole that the present invention synthesizes-mesoporous silica gel integral post tool and microstructure, lead to
Permeability is strong, and back pressure is low, and mass transfer dynamicses are fast.
(2) it is located near or at polymerizeing using surface imprinted technology synthesis bovine serum albumin(BSA) imprinted polymer, recognition site
Thing surface, facilitate bovine serum albumin(BSA) molecule to pass in and out imprinted cavity, improve the selectivity to bovine serum albumin(BSA).
(3) stratum's ordered big hole-mesoporous silica gel integral post bovine serum albumin(BSA) imprinted polymer has to bovine serum albumin(BSA)
Good separating property, adsorption capacity is up to 121.6mg/g.
Brief description of the drawings
Fig. 1 is that polystyrene colloid crystal (curve a), impregnates the polystyrene colloid after precursor solution in embodiment 1
Crystal (curve b), HOM (curve c) and BSA-MIP (curve d) infrared spectrums.
The macropore (figure a) that Fig. 2 is HOM in embodiment 1, mesoporous (figure b) structure and BSA-MIP (figure c) transmission electron microscope picture.
Embodiment
With reference to specific implementation example, the present invention will be further described.
Embodiment 1：
(1) 0.4g polystyrene is dissolved in 8mL water, by solution ultrasound 20min, natural subsidence one week, carefully removes upper strata
Liquid, 25 DEG C of vacuum drying 48h, obtains polystyrene colloid crystal.
(2) 2g TEOS, 0.9g water, 0.8g hydrochloric acid, 1.0g P123 and 15g absolute ethyl alcohol, which are prepared, turns into precursor solution,
0.4g polystyrene colloid crystal is added in solution, impregnates 1h, wherein concentration of hydrochloric acid is 2mol/L.Then in 30 DEG C of dippings
Evaporate 48h.The bulk material of synthesis is placed in tube furnace, in atmosphere with 1 DEG C of min-1Programming rate slowly heat up, extremely
Constant temperature 6h at 550 DEG C, obtain stratum's ordered big hole-mesoporous silica gel integral post (HOM).
(3) HOM of gained is dissolved in 40mL absolute ethyl alcohols in 0.4g steps (2).Then 2mL MPS are taken to be added to mixing molten
In liquid, 40 DEG C of reaction 12h.Product is washed 3 times with absolute ethyl alcohol after reaction, to elute unreacted MPS, 50 DEG C of vacuum drying
Obtain the HOM (HOM@MPS) of MPS modifications.
(4) the HOM@MPS that 0.05g steps (3) obtain are dissolved in the 20mmol/L phosphate buffers of 20mL pH 7.0, are surpassed
Sound 10min, 35.5mg AM, 77mg MBA, 20mg BSA are added in solution, 35 DEG C of stirring 1h, lead to N2, by 10mg APS
It is added to 10 μ L TEMED in solution, after 35 DEG C of reaction 24h reactions, washs 5 times repeatedly with deionized water to remove unreacted
AM, APS and BSA, 25 DEG C vacuum drying.Then with the second of lauryl sodium sulfate (SDS) of the 1000mL containing 100mg/L
Acid：Water (v:v,1:9) solution washes away BSA template molecules, and is washed with deionized water to neutrality, and 25 DEG C are dried in vacuo and produce stratum and have
The foramen magnum-mesoporous monolithic silica column bovine serum albumin(BSA) imprinted polymer (BSA-MIP) of sequence.
Fig. 1 is that polystyrene colloid crystal (curve a), impregnates the polystyrene colloid after precursor solution in the present embodiment
Crystal (curve b), HOM (curve c) and BSA-MIP (curve d) infrared spectrums.As can be seen from the figure after calcining,
The peak of polystyrene is not seen, illustrates that polystyrene colloid crystal template removes completely.After polymerisation, in 1730cm-1Place
There is a new spike to occur, this is the characteristic peak of carboxyl, shows that imprinted polymer successfully adheres to.
The macropore (figure a) that Fig. 2 is HOM in the present embodiment, mesoporous (figure b) structure and BSA-MIP (figure c) transmission electron microscope
Figure.As seen from the figure, stratum's ordered big hole-mesoporous silica gel integral post bovine serum albumin(BSA) imprinted polymer by silica skeleton and
Macropore is formed, and has on silica skeleton equally distributed mesoporous, and stratum's ordered big hole-mesoporous silica gel integral post tool is well-regulated
The macroporous structure to communicate with one another, bore dia are 1~1.2 μm.There are homogeneous meso-hole structure, aperture 6-7nm again on skeleton.It is logical
Cross comparison diagram c and figure b, it can be seen that come, after polymerisation, one layer of bovine serum albumin(BSA) trace is formd on the surface of integral post and is gathered
Compound layer.
Embodiment 2：
(1) 0.8g polystyrene is dissolved in 8mL water, by solution ultrasound 20min, natural subsidence one week, carefully removes upper strata
Liquid, 25 DEG C of vacuum drying 48h, obtains polystyrene colloid crystal.
(2) 1.5g TEOS, 0.9g water, 0.1g hydrochloric acid, 1.0g P123 and 15g absolute ethyl alcohol are prepared molten as presoma
Liquid, 0.6g polystyrene colloid crystal is added in solution, impregnates 1h, wherein concentration of hydrochloric acid is 2mol/L.Then at 30 DEG C
Dipping evaporation 48h.The bulk material of synthesis is placed in tube furnace, in atmosphere with 1 DEG C of min-1Programming rate slowly rise
Temperature, to constant temperature 6h at 550 DEG C, obtain stratum's ordered big hole-mesoporous silica gel integral post (HOM).
(3) HOM of gained is dissolved in 40mL absolute ethyl alcohols in 0.4g steps (2).Then 2mL MPS are taken to be added to mixing molten
In liquid, 40 DEG C of reaction 12h.Product is washed 3 times with absolute ethyl alcohol after reaction, to elute unreacted MPS, 50 DEG C of vacuum drying
Obtain the HOM (HOM@MPS) of MPS modifications.
(4) the HOM@MPS that 0.08g steps (3) obtain are dissolved in the 20mmol/L phosphate buffers of 20mL pH 7.5, are surpassed
Sound 10min, 71.0mg AM, 77mg MBA, 10mg BSA are added in solution, 35 DEG C of stirring 1h, lead to N2, by 8mg APS and
10 μ L TEMED are added in solution, 35 DEG C reaction 24h reaction after, washed repeatedly with deionized water 5 times it is unreacted to remove
AM, APS and BSA, in 25 DEG C of vacuum drying.Then with the acetic acid of lauryl sodium sulfate (SDS) of the 750mL containing 100mg/L：Water
(v:v,1:9) solution washes away BSA template molecules, and is washed with deionized water to neutrality, and 25 DEG C are dried in vacuo that to produce stratum big in order
Hole-mesoporous silica gel integral post bovine serum albumin(BSA) imprinted polymer (BSA-MIP).
Embodiment 3：
(1) 1.0g polystyrene is dissolved in 8mL water, by solution ultrasound 20min, natural subsidence one week, carefully removes upper strata
Liquid, 25 DEG C of vacuum drying 48h, obtains polystyrene colloid crystal.
(2) 2.5g TEOS, 0.9g water, 0.4g hydrochloric acid, 1.0g P123 and 15g absolute ethyl alcohol are prepared molten as presoma
Liquid, 1.0g polystyrene colloid crystal is added in solution, impregnates 1h, wherein concentration of hydrochloric acid is 2mol/L.Then at 30 DEG C
Dipping evaporation 48h.The bulk material of synthesis is placed in tube furnace, in atmosphere with 1 DEG C of min-1Programming rate slowly rise
Temperature, to constant temperature 6h at 550 DEG C, obtain stratum's ordered big hole-mesoporous silica gel integral post (HOM).
(3) HOM of gained is dissolved in 40mL absolute ethyl alcohols in 0.4g steps (2).Then 2mL MPS are taken to be added to mixing molten
In liquid, 40 DEG C of reaction 12h.Product is washed 3 times with absolute ethyl alcohol after reaction, to elute unreacted MPS, 50 DEG C of vacuum drying
Obtain the HOM (HOM@MPS) of MPS modifications.
(4) the HOM@MPS that 0.10g steps (3) obtain are dissolved in the 20mmol/L phosphate buffers of 20mL pH 8.0, are surpassed
Sound 10min, 50.0mg AM, 77mg MBA, 15mg BSA are added in solution, 35 DEG C of stirring 1h, lead to N2, by 9mg APS and
10 μ L TEMED are added in solution, 35 DEG C reaction 24h reaction after, washed repeatedly with deionized water 5 times it is unreacted to remove
AM, APS and BSA, in 25 DEG C of vacuum drying.Then with the acetic acid of lauryl sodium sulfate (SDS) of the 500mL containing 100mg/L：Water
(v:v,1:9) solution washes away BSA template molecules, and is washed with deionized water to neutrality, and 25 DEG C are dried in vacuo that to produce stratum big in order
Hole-mesoporous silica gel integral post bovine serum albumin(BSA) imprinted polymer (BSA-MIP).
Claims (6)
- A kind of 1. stratum's ordered big hole-mesoporous silica gel integral post bovine serum albumin(BSA) imprinted polymer, it is characterised in that the stratum Ordered big hole-mesoporous silica gel integral post bovine serum albumin(BSA) imprinted polymer is made up of silica skeleton and macropore, in silica Have on skeleton equally distributed mesoporous；The macropore diameter is 1 ~ 1.2 μm, and the structure interconnected is formed between macropore, described Mesoporous pore size is 6 ~ 7nm；Stratum's ordered big hole-mesoporous silica gel integral post bovine serum albumin(BSA) imprinted polymer is to cow's serum The adsorption capacity of albumin is up to 121.6mg/g.
- 2. a kind of preparation method of stratum's ordered big hole-mesoporous silica gel integral post bovine serum albumin(BSA) imprinted polymer, its feature It is, comprises the following steps：The synthesis of step 1, polystyrene colloid crystal：Polystyrene is dissolved in a certain amount of water, the dosage of polystyrene and water Than for 0.4 ~ 1g：8mL；By solution ultrasound 20min, then natural subsidence one week, carefully removes supernatant liquid, 25 DEG C of vacuum drying 48h, obtain polystyrene colloid crystal；The synthesis of step 2, stratum's ordered big hole-mesoporous silica gel integral post：A certain amount of tetraethyl orthosilicate, water, hydrochloric acid, gather Poly- (propane diols) block of (1,2- ethylidene glycol) block poly- (1,2- ethylidene glycol) and absolute ethyl alcohol, which are prepared, turns into presoma Solution, polystyrene colloid crystal is added in solution, impregnates 1h, obtain mixed liquor A, wherein concentration of hydrochloric acid is 2mol/L； Then by mixed liquor A in 30 DEG C of dipping evaporation 48h；The bulk material of synthesis is placed in tube furnace, in atmosphere with 1 DEG C min-1Programming rate slowly heat up, to constant temperature 6h at 550 DEG C, obtain stratum's ordered big hole-mesoporous silica gel integral post, referred to as HOM；Step 3, stratum's ordered big hole-mesoporous silica gel integral post are modified with γ-(methacryloxypropyl) propyl trimethoxy silicane It is prepared by material：The HOM of gained is dissolved in 40mL absolute ethyl alcohols in 0.4g steps 2, then takes 2mL γ-(methacryloxypropyl) third Base trimethoxy silane (MPS) is added in mixed solution, 40 DEG C of reaction 12h；Product is washed 3 with absolute ethyl alcohol after reaction Secondary, to elute unreacted MPS, 50 DEG C of vacuum drying obtain the HOM, referred to as HOM@MPS of MPS modifications；The preparation of step 4, stratum's ordered big hole-mesoporous silica gel integral post bovine serum albumin(BSA) imprinted polymer (BSA-MIP)：Step Rapid 3 obtained HOM@MPS are scattered in phosphate buffer, ultrasonic 10min, mixed liquid B are obtained, by acrylamide, N, N '-sub- Bisacrylamide, bovine serum albumin(BSA) are added in mixed liquid B, obtain mixed liquor C, and 1h is stirred at 35 DEG C to mixed liquor C, Logical N2, by ammonium persulfate and N, N, N, N- tetramethylethylenediamines are added in solution, 35 DEG C of reaction 24h；After reaction, deionization is used Water washs 5 times to remove unreacted AM, APS and BSA repeatedly, in 25 DEG C of vacuum drying；Then contained with 500 ~ 1000mL The acetic acid of 100mg/L lauryl sodium sulfate：The aqueous solution washes away BSA template molecules, and is washed with deionized water to neutrality, 25 DEG C It is dried in vacuo and produces stratum's ordered big hole-mesoporous silica gel integral post bovine serum albumin(BSA) imprinted polymer.
- A kind of 3. stratum's ordered big hole according to claim 2-mesoporous silica gel integral post bovine serum albumin(BSA) trace polymerization The preparation method of thing, it is characterised in that in step 2, when preparing mixed liquor A, tetraethyl orthosilicate used, water, hydrochloric acid, gather Poly- (propane diols) block of (1,2- ethylidene glycol) block is poly- (1,2- ethylidene glycol), and absolute ethyl alcohol and polystyrene colloid are brilliant The mass ratio of body is 1.5 ~ 2.5g：0.9g：0.1~0.8g：1.0g：15g：0.4~1g.
- A kind of 4. stratum's ordered big hole according to claim 2-mesoporous silica gel integral post bovine serum albumin(BSA) trace polymerization The preparation method of thing, it is characterised in that in step 4, when preparing mixed liquid B, HOM@MPS concentration is 2.5 ~ 5mg/mL；Prepare During mixed liquor C, HOM@MPS used, acrylamide, N, N '-methylene-bisacrylamide, bovine serum albumin(BSA), ammonium persulfate, The amount ratio of N, N, N, N- tetramethylethylenediamine is 0.05 ~ 0.10g：35.5~71.0mg：77mg：10~20mg：8~10mg：10μ L。
- A kind of 5. stratum's ordered big hole according to claim 2-mesoporous silica gel integral post bovine serum albumin(BSA) trace polymerization The preparation method of thing, it is characterised in that in step 4, the acetic acid of the lauryl sodium sulfate containing 100mg/L：The aqueous solution In, the volume ratio of acetic acid and water is 1：9.
- A kind of 6. stratum's ordered big hole according to claim 2-mesoporous silica gel integral post bovine serum albumin(BSA) trace polymerization The preparation method of thing, it is characterised in that the phosphate buffering liquid concentration described in step 4 is 20mmol/L, and pH is 7.0 ~ 8.0.
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