CN104624167B - The preparation of the integral material of immobilized aptamer and the application in albumen on-line checking - Google Patents
The preparation of the integral material of immobilized aptamer and the application in albumen on-line checking Download PDFInfo
- Publication number
- CN104624167B CN104624167B CN201310554485.4A CN201310554485A CN104624167B CN 104624167 B CN104624167 B CN 104624167B CN 201310554485 A CN201310554485 A CN 201310554485A CN 104624167 B CN104624167 B CN 104624167B
- Authority
- CN
- China
- Prior art keywords
- aptamer
- integral material
- preparation
- mixture
- sulfydryl
- 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
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
Abstract
The present invention relates to the preparation and its application in protein on-line checking of a kind of aptamer modified affine integral material, belong to analysis technical field.The integral material of described immobilized aptamer is hydrophilic polymer obtained in by way of in-situ polymerization.Then using the epoxide group of stromal surface, using chemical derivatization, stromal surface is made rich in sulfydryl, so as to modified nano gold granule；The aptamer of the last nanometer gold surface covalent bonding sulfydryl modification in substrate, and based on aptamer and the specific recognition feature detection target protein of target protein.This is affine, and integral material can be used for the on-line preconcentration and Sensitive Detection of protein.The method has high selectivity and the good response rate.
Description
Technical field
The present invention relates to aptamer is used for the enrichment and detection of albumen, specifically a kind of aptamer modified parent
With preparing and its for the efficient high-selectivity enrichment and online Sensitive Detection of target protein for integral material.
Background technology
Aptamer（Aptamer, also known as aptamers, aptamer）It is energy high-affinity, combines target with high specificity
Single stranded oligonucleotide（ssDNA or ssRNA）.Aptamer can pass through part index concentration phyletic evolution technology
（Systematic Evolution of Ligands by EXponential enrichment, SELEX）Screening is obtained, and is borrowed
Van der Waals force, hydrogen bond, pi-pi accumulation, hydrophobic interaction equimolecular intermolecular forces are helped to form special three dimensional structure, so as to specifically know
Other target.Screening obtains aptamer and has that affinity is good, selectivity is high, it is stable in properties, many advantages, such as be easy to chemical modification,
The aptamer target reported at present includes metal ion, small molecule, polypeptide, protein, cell, and application is wide.
Integral post is relative to open tubular column and packed column, little with back-pressure, and permeability is good, it is easy to accomplish the advantage such as online, its
Using widely.Reported success is had at present by aptamer modified on integral material, and realizing has to target protein
Effect enrichment（Anal.Chem.2001,73,5415-5421;J.Chromatogr.A2006,1111,115-119;
J.Chromatogr.B2012,903,112-117;Anal.Chem.2012,84,10186-10190）.So far it is suitable for nucleic acid
The integral material of body is mainly divided to two classes：Polymeric matrix and silica gel material substrate.Wherein, the integral material table based on silica matrix
, there is stronger non-specific adsorption to protein in the silicone hydroxyl in face, can affect the selectivity being enriched with, and the material of silica matrix
Tolerable pH narrow ranges；And can have preferable stability, and polyethyleneglycol diacrylate using polymeric matrix
Non-specific adsorption can be reduced with preferable hydrophilic；In addition, immobilized nanogold particle being capable of stably covalent bond mercapto
The aptamer of base modification, while also having hydrophilic, it is possible to increase the detection response rate to target protein.
The content of the invention
Aptamer is single-stranded oligonucleotide（ssDNA or ssRNA）, special three dimensional structure can be formed, to target
Mark is with high selectivity and high specific, and stability is high, it is easy to chemical modification.Aptamer is immobilized on into hydrophilic modifying
Non-specific adsorption can be reduced in integral material, the response rate of detection is improved.It is an object of the invention to provide a kind of good hydrophilic property
Integrated substrate carrier, the immobilized nanometer gold in surface, the further aptamer of immobilized sulfydryl modification, so as to prepare non-specific suction
The good affine integral post of attached primary school, the response rate, for the high power capacity of protein, high-selectivity enrichment and highly sensitive on-line checking.
For achieving the above object, the technical solution used in the present invention is：
Using glycidyl methacrylate and polyethyleneglycol diacrylate as monomer, enter in quartz capillary
Row in-situ thermo-polymerization, obtains hydrophilic polymer integrated substrate, and Jing epoxy ring opening reactions introduce sulfydryl on polymeric matrix surface,
Then in the immobilized nanogold particle in its surface, further in the aptamer of nanogold particle surface covalent bond sulfydryl modification.
The preparation of the aptamer modified affine integral material, comprises the following steps that：
（1）The preparation of the hydrophilic integral material substrate containing epoxide group：With glycidyl methacrylate（Quality point
Number is 20-30%（w/w））And polyethyleneglycol diacrylate（Mass fraction is 10-20%（w/w））As monomer, pore is added
Agent normal propyl alcohol and cyclohexanol mixture（Mass fraction is 50-70%（w/w））, and initiator azodiisobutyronitrile（Mass fraction
Account for the 0.05-1% of monomer and porogen gross mass（w/w））.Concussion ultrasound is mixed, and is led to nitrogen and is removed dissolved oxygen, fills post, and heating is poly-
Unreacted reagent is gone out with pure methanol and pure water respectively after closing 10-24 hours；
（2）Stromal surface modified nano gold：Half is passed through in the prepared capillary monolithic column containing epoxide group continuously
Cystamine（CD）Solution, reacts 6 hours at 40-70 DEG C, using 1M Tris-HCl（pH=8）After buffer solution sealing 2-4 hours
With pure water rinsing pillar to neutrality, tricarboxylic methyl acid phosphate is continuously passed through at room temperature（TCEP）Solution reduction disulfide bond, Ran Houyong
Pure water rinsing pillar；Most concentration 2-10nM nano-Au solution 1.5-4mL is passed through in the integral material substrate rich in sulfydryl backward；
（3）The preparation of the integral material of the immobilized aptamer of core：It is continuous in the integral material substrate for modified nanometer gold
The Tris-HCl buffer solution of the aptamer containing sulfydryl modification is passed through, under room temperature, 1-48h is reacted.
The integral material of the immobilized aptamer for preparing can be used for optionally on-line preconcentration and detect that aptamer can be caught
Catch target protein.
The invention has the advantages that：
1. the present invention adopts aptamer to the affine enrichment of target, with high selectivity and high specific；
2. the present invention makees cross-linking agent using the hydrophilic monomer polyethyleneglycol diacrylate containing polyglycol chain, preparation
Host material good hydrophilic property, non-specific adsorption are little.
3. the present invention adopts nanometer gold integral material, can stably be bonded the aptamer of sulfydryl modification, excellent hydrophilic,
On-line preconcentration is easily achieved, the good response rate is obtained in that.
4. the material stiffness for preparing, mechanical strength are good, can tolerate the pressure limit of 0-40MPa, applicable pH range width（1-
14）.
Description of the drawings
Fig. 1 is the preparation flow figure of aptamer modified affine integral material；
Fig. 2 is the affine integral post of Thrombin aptamer modification to myoglobon（Myo）、BSA、transferrin
（TransFe）And thrombin（Thrombin）The liquid chromatograph stacking chart of detection；
Fig. 3 is the affine integral post for verifying Thrombin aptamer modification to myoglobon（Myo）、BSA、
transferrin（TransFe）And thrombin（Thrombin）The SDS-PAGE figures of reservation situation；
Fig. 4 is the liquid chromatograph stacking chart that the affine integral post of Thrombin aptamer modification is detected to thrombin；
Fig. 5 is linear equation of the affine integral post of Thrombin aptamer modification to thrombin detection by quantitative；
Fig. 6 is the standard protein sample liquid chromatograph stacking chart for detecting the thrombin response rate；
Fig. 7 is the linear equation for detecting the thrombin response rate；
Fig. 8 is result of calculation of the affine integral post of Thrombin aptamer modification to the thrombin response rate；
Fig. 9 is testing result of the affine integral post of Thrombin aptamer modification to thrombin in complex sample.
Specific embodiment
The method that the present invention is provided is described in detail below by embodiment, but the invention is not limited in any way.
Embodiment 1
As shown in figure 1, the affine integral material of Thrombin aptamer modification is prepared by following flow process：
1）The preparation of the substrate of integral material containing epoxide group：It is formulated as follows pre-gathering solutions：240mg Glycidyl methacrylates
Glyceride, 160mg polyethyleneglycol diacrylates, 50mg normal propyl alcohols, 550mg Hexalin and 4mg azodiisobutyronitriles.By pre-polymerization
Solution removes the dissolved oxygen in solution with nitrogen, then ultrasound 10min in ice-water bath；Import the capillary tube through double bond modification
It is interior, sealing, and at 50 DEG C cause polymerization, polymerization time after 12 hours respectively with pure methanol and pure water go out unreacted monomer and
Porogen；
2）Stromal surface modified nano gold：1M half is passed through in the prepared capillary monolithic column containing epoxide group continuously
Cystamine solution（Dissolved using 1M NaOH solutions）, react 6 hours at 50 DEG C, using 1M Tris-HCl（pH=8.0）Buffering is molten
0.25M tricarboxylic methyl acid phosphate solution is at room temperature continuously passed through to neutrality with pure water rinsing pillar after fluid-tight tail 2 is little（Use quality
Concentration 28-30% ammonia adjusts pH to neutrality）Reduction of Disulfide, uses pure water rinsing pillar；Most backward in the integral material rich in sulfydryl
It is passed through homemade concentration 8nM nano-Au solution（About 15nm）2mL, reddening to effluent, supported quantity about 16pmol；
3）The preparation of the affine integral material of Thrombin aptamer modification：Connect in the affine integral material of above-mentioned preparation
It is continuous to be passed through the Thrombin aptamer containing sulfydryl modification（Purchased from precious biological engineering (Dalian) company limited）Tris-HCl buffering
Solution（pH=7.4）, under room temperature, react 4h.By spacerarm between mercapto groups and aptamer（6 methylene）Connection.
Embodiment 2
As shown in figure 1, the affine integral material of Thrombin aptamer modification is prepared by following flow process：
1）The preparation of the substrate of integral material containing epoxide group：It is formulated as follows pre-gathering solutions：240mg Glycidyl methacrylates
Glyceride, 160mg polyethyleneglycol diacrylates, 400mg normal propyl alcohols, 200mg Hexalin and 4mg azodiisobutyronitriles.Will be pre-
Poly solution removes the dissolved oxygen in solution with nitrogen, then ultrasound 10min in ice-water bath；Import the capillary tube through double bond modification
It is interior, sealing, and at 50 DEG C cause polymerization, polymerization time after 12 hours respectively with pure methanol and pure water go out unreacted monomer and
Porogen；
2）Stromal surface modified nano gold：1M half is passed through in the prepared capillary monolithic column containing epoxide group continuously
Cystamine solution（Dissolved using 1M NaOH solutions）, react 6 hours at 50 DEG C, using 1M Tris-HCl（pH=8.0）Buffering is molten
0.25M tricarboxylic methyl acid phosphate solution is at room temperature continuously passed through to neutrality with pure water rinsing pillar after fluid-tight tail 2 is little（Use quality
Concentration 28-30% ammonia adjusts pH to neutrality）Reduction of Disulfide, uses pure water rinsing pillar；Most backward in the integral material rich in sulfydryl
It is passed through homemade concentration 8nM nano-Au solution（About 15nm）4mL, reddening to effluent, supported quantity about 32pmol；3）Blood coagulation
The preparation of the aptamer modified affine integral material of enzymatic nucleic acid：Continuously it is passed through in the affine integral material of above-mentioned preparation and repaiies containing sulfydryl
The Thrombin aptamer of decorations（Purchased from precious biological engineering (Dalian) company limited）Tris-HCl buffer solution（pH=7.4）, room
The lower reaction 4h of temperature.By spacerarm between mercapto groups and aptamer（18 methylene）Connection.
Embodiment 3
The selectivity of the affine integral post to thrombin of Thrombin aptamer modification, concrete determination step is：
1）On liquid chromatographic system（With the affine integral material that Thrombin aptamer prepared by embodiment 1 is modified it is
Separation material）, using in Tris-HCl buffer（Containing 20mM Tris-HCl, 140mM NaCl, 5mM KCl, 1mM
MgCl2,1mM CaCl2,pH=7.4）As sample-loading buffer, the myoglobin of loading 200ng/ μ L successively（Myo）、BSA、
transferrin（TransFe）And thrombin（Thrombin）Each 2 μ L, use 2M NaClO4As eluting solution, as a result such as Fig. 2
It is shown.Collect the not retained fraction and elution fraction of the not retained fraction and Thrombin of Myo, BSA, TransFe.
2）Above-mentioned sample is carried out into SDS-PAGE analyses：It is with freezer dryer by above-mentioned sample lyophilizing, each to add on 20 μ L
Sample buffer weight is molten.20 μ L Myo, BSA, TransFe and Thrombin standard protein samples are taken again respectively in addition.To all samples
The middle SDS-PAGE loading buffers for adding equal volume, after boiling 5min, are added in gel duct in boiling water.Adopt
With 5% concentration glue and 12% separation gel, separated under 120V constant-pressure conditions.As a result as shown in figure 3, target protein
The elution fraction of Thrombin is consistent with the band of Thrombin standard proteins, and retained fraction does not find any band；
It is for the not retained fraction of non-targeted albumen Myo, BSA and TransFe, consistent with the band of respective standard protein.This shows,
The affine integral post of Thrombin aptamer modification can exclusively recognize thrombin, with extraordinary selectivity.
Embodiment 4
Power of test of the affine integral post of Thrombin aptamer modification to thrombin（With blood coagulation prepared by embodiment 1
The aptamer modified affine integral material of enzymatic nucleic acid is separation material）, concretely comprise the following steps：
Loading 0.5 successively, each 2 μ L of 2.5,5,25,50,200,300ng/ μ L, same time 2M NaClO4Washed
It is de-.As a result it is as shown in Figure 4.The peak area for retaining peak is calculated, the meansigma methodss of 3 tests is taken, figure, linear side is done to applied sample amount
Journey, as a result as shown in Figure 5.As a result show, the affine integral post of Thrombin aptamer modification to the detection range of thrombin is
1-600ng, with the good range of linearity, and detection by quantitative is limited to 1ng（13.6nM）, qualitative detection is limited to 0.5ng
（6.8nM）.
Embodiment 5
The response rate that the affine integral post of Thrombin aptamer modification is detected to thrombin, concretely comprises the following steps：
With A phases（95% water+5%ACN+0.1%TFA, v/v/v）For loading mobile phase, B phases（5% water+95%ACN+0.1%TFA,
v/v/v）For elution flow phase, the anti-phase capillary packed columns of C4 are detected.Loading 25,50,100,150,200ng/ μ L successively
The each 2 μ L of thrombin standard albumen, carry out eluting with the ACN of final concentration 80%, as a result as shown in Figure 6.Calculate each and retain peak
Peak area, does figure to applied sample amount, linear equation, as a result as shown in Figure 7.It is affine whole by what is modified from Thrombin aptamer
The 3 parts of samples for eluting on scapus and collecting are sequentially entered in system, calculate respective eluting peak area, as a result such as Fig. 8 institutes
Show.This shows that the response rate that the affine integral post of Thrombin aptamer modification is detected to thrombin is 92.6 ± 4.5%（RSD=
4.8%）, the response rate is good.
Embodiment 6
Power of test of the affine integral post of Thrombin aptamer modification to thrombin in complex sample（With embodiment 1
The affine integral material of the Thrombin aptamer modification of preparation is separation material）, concretely comprise the following steps：
1. protein mixed solution is prepared
By thrombin（71ng/μL）With human serum sample（71μg/μL）By concentration ratio 1:1000 mixing, obtain albumen mixing
Solution（35.54μg/μL）.In addition, by human serum sample（71μg/μL）1 times is diluted with the sample-loading buffer of same volume, obtain dilute
The human serum sample for releasing（35.50μg/μL）.
2. selective enrichment
Loading dilutes successively human serum sample and each 2 μ L of protein mixed solution, same time 2M NaClO4Washed
It is de-, as a result as shown in Figure 9.Protein mixed solution in figure（1:1000）Eluting peak apparently higher than dilution human serum sample
（serum）, this shows 1:In 1000 complex sample system, the affine integral post of Thrombin aptamer modification still can be caught
Collect target protein thrombin.
Claims (6)
1. the integral material of immobilized aptamer, it is characterised in that：Aptamer using sulfydryl modification is common with integral material
The nanogold particle of valency modification is combined, and forms the integral material of immobilized aptamer；The integral material of preparation, adopts and contains epoxy
The methyl acrylic ester compound and diacrylate esters compound of group is monomer, using volume ratio 1:12-12:1 just
The mixture of propanol and Hexalin is used as porogen；
Wherein, count in mass ratio, the methyl acrylic ester compound containing epoxide group accounts for the 20%-30% of total mixture,
Diacrylate esters compound accounts for the 10%-20% of mixture, and porogen accounts for the 50%-70% of mixture；The addition of initiator
Measure the 0.05%-1% for total mixture；
Nanogold particle size is 2-50nm.
2. the preparation method of the integral material of the immobilized aptamer described in a kind of claim 1, it is characterised in that：Using containing
The methyl acrylic ester compound and diacrylate esters compound of epoxide group as reaction monomers, in quartz capillary
In-situ thermo-polymerization is carried out, the hydrophilic polymer integrated substrate containing epoxide group is obtained, Jing epoxy ring opening reactions are in polymer
Stromal surface introduces sulfydryl, then in its surface finish nano gold grain, further repaiies in nanometer gold surface covalent bond sulfydryl
The aptamer of decorations；
Using volume ratio 1:12-12:1 normal propyl alcohol and the mixture of Hexalin are used as porogen；
Wherein, count in mass ratio, the methyl acrylic ester compound containing epoxide group accounts for the 20%-30% of total mixture,
Diacrylate esters compound accounts for the 10%-20% of mixture, and porogen accounts for the 50%-70% of mixture；The addition of initiator
Measure the 0.05%-1% for total mixture；
Nanogold particle size is 2-50nm.
3. according to the preparation method described in claim 2, it is characterised in that：Hydrophilic polymer containing epoxide group entirety material
Material substrate adopts glycidyl methacrylate and polyethyleneglycol diacrylate for monomer, by methyl propenoic acid glycidyl
After ester, polyethyleneglycol diacrylate, porogen mix homogeneously total mixture, total mixture is in quartz capillary by causing
Agent causes in-situ thermo-polymerization to form.
4. according to the preparation method described in claim 2, it is characterised in that：The feature of described introducing sulfydryl is：To containing ring
Cysteamine (CD) solution is continuously passed through in the hydrophilic polymer integral material substrate of oxygen groups to be reacted；Use pH=7.5-
With pure water rinsing pillar to neutrality after 10.0 Tris-HCl buffer solution sealing 2-4h；Then continuously it is passed through tricarboxylic methyl acid phosphate
(TCEP) solution reduction disulfide bond；Again with concentration 2-10nM nano-Au solution, nanometer gold supported quantity are passed through after pure water rinsing pillar
For 3-40pmol；
Cysteamine (CD) solution is that cysteamine (CD) is dissolved using 1-2M NaOH solutions, and its concentration is 0.5-2M；Reaction temperature
For 40-70 DEG C, the response time is 1-12h；
Tricarboxylic methyl acid phosphate (TCEP) solution adjusts pH to neutrality with the ammonia of mass concentration 28-30%, and concentration is 0.1-1M, reduction
The temperature of disulfide bond is 15-50 DEG C, and the time is 1-12h.
5. according to the preparation method described in claim 2, it is characterised in that：
Continuously it is passed through the pH=7.2-8.0's of the aptamer containing sulfydryl modification in the integral material for modified nanometer gold
1-48h is reacted under Tris-HCl buffer solution, room temperature；
The aptamer of sulfydryl modification is dissolved in the Tris-HCl buffer of pH=7.2-8.0, the aptamer of sulfydryl modification
Molar concentration be 1-500 μM；Connected every arm by conduct between 2-18 methylene between mercapto groups and aptamer.
6. a kind of integral material of the immobilized aptamer described in claim 1 is used for efficient, online sensitive inspection with high selectivity
Survey the seizable target protein of aptamer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310554485.4A CN104624167B (en) | 2013-11-08 | 2013-11-08 | The preparation of the integral material of immobilized aptamer and the application in albumen on-line checking |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310554485.4A CN104624167B (en) | 2013-11-08 | 2013-11-08 | The preparation of the integral material of immobilized aptamer and the application in albumen on-line checking |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104624167A CN104624167A (en) | 2015-05-20 |
| CN104624167B true CN104624167B (en) | 2017-04-05 |
Family
ID=53203691
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310554485.4A Active CN104624167B (en) | 2013-11-08 | 2013-11-08 | The preparation of the integral material of immobilized aptamer and the application in albumen on-line checking |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104624167B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106442796B (en) * | 2016-10-26 | 2019-12-03 | 复旦大学 | A kind of liquid phase open tubular column of multi-layer nano gold goal and preparation method thereof and application |
| CN110314673B (en) * | 2019-08-08 | 2021-06-01 | 福州大学 | Aptamer functionalized affinity monolithic column based on photo-initiated hybrid polymerization and preparation method thereof |
| EP4023682A4 (en) * | 2019-08-30 | 2023-11-08 | Canon Kabushiki Kaisha | Particles, affinity particles having ligand for target substance, in vitro diagnostic reagent and kit that include same, and method for detecting target substance |
| CN113333042B (en) * | 2021-06-21 | 2022-04-22 | 太原理工大学 | Micro-fluidic chip for nucleic acid detection and manufacturing method thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102114414A (en) * | 2009-12-30 | 2011-07-06 | 中国科学院大连化学物理研究所 | Method for preparing immobilized metal ion affinity chromatographic monolithic column |
| CN102766005A (en) * | 2012-08-01 | 2012-11-07 | 福州大学 | Chiral compound separation method based on nano gold modified by aptamer |
-
2013
- 2013-11-08 CN CN201310554485.4A patent/CN104624167B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102114414A (en) * | 2009-12-30 | 2011-07-06 | 中国科学院大连化学物理研究所 | Method for preparing immobilized metal ion affinity chromatographic monolithic column |
| CN102766005A (en) * | 2012-08-01 | 2012-11-07 | 福州大学 | Chiral compound separation method based on nano gold modified by aptamer |
Non-Patent Citations (2)
| Title |
|---|
| Aptamer-Functionalized Au Nanoparticles for the Amplified Optical Detection of Thrombin;Valeri Pavlov et al.;《J. AM. CHEM. SOC.》;20040309;第126卷;第11768-11769页 * |
| Preparation of porous polymer monoliths featuring enhanced surface coverage with gold nanoparticles;Yongqin Lv et al.;《Journal of Chromatography A》;20120411;第1261卷;第121-128页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104624167A (en) | 2015-05-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Masini et al. | Porous monoliths for on-line sample preparation: a review | |
| Du et al. | Recent advances in aptamer-functionalized materials in sample preparation | |
| Li et al. | Boronate affinity materials for separation and molecular recognition: structure, properties and applications | |
| Wilson et al. | Nano liquid chromatography columns | |
| Safdar et al. | Microscale immobilized enzyme reactors in proteomics: latest developments | |
| Yue et al. | Ultratrace LC/MS proteomic analysis using 10-μm-id porous layer open tubular poly (styrene− divinylbenzene) capillary columns | |
| CN104624167B (en) | The preparation of the integral material of immobilized aptamer and the application in albumen on-line checking | |
| Andaç et al. | Affinity-recognition-based polymeric cryogels for protein depletion studies | |
| US20040126890A1 (en) | Biomolecule open channel solid phase extraction systems and methods | |
| US8399055B2 (en) | Open channel solid phase extraction systems and methods | |
| Gunasena et al. | Organic monoliths for hydrophilic interaction electrochromatography/chromatography and immunoaffinity chromatography | |
| JP2015512621A5 (en) | ||
| Krenkova et al. | Less common applications of monoliths: V. Monolithic scaffolds modified with nanostructures for chromatographic separations and tissue engineering | |
| Nevídalová et al. | Capillary electrophoresis–based immunoassay and aptamer assay: A review | |
| Wang et al. | Hydrophilic polymeric monoliths containing choline phosphate for separation science applications | |
| Yao et al. | Protein adsorption in supermacroporous cryogels with embedded nanoparticles | |
| Ying et al. | Poly (glycidyl methacrylate) nanoparticle-coated capillary with oriented antibody immobilization for immunoaffinity in-tube solid phase microextraction: Preparation and characterization | |
| Pichon et al. | Immunosorbents in microextraction | |
| Pichon | Aptamer-based and immunosorbents | |
| CN114740055B (en) | Linear range adjustable accurate immunosensory method and portable biological resistance sensing measurement device | |
| CN113976195A (en) | Microfluidic chip for exosome separation and enrichment and method for analyzing exosome surface protein | |
| CN103333299B (en) | Glycidyl methacrylate and PEG alkylmethacrylate polymer microballoon and its preparation method and application | |
| CN104606919B (en) | Preparation method of nucleic acid aptamer/nano-gold modified solid phase microextraction quartz capillary tube | |
| Bie et al. | Selective analysis of interferon-alpha in human serum with boronate affinity oriented imprinting based plastic antibody | |
| Suo et al. | Monolithic capillary microextraction on-line combined with ICP-MS for determining Ni, Cu and Cd in biological samples |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |