CN110511935A - A kind of aptamers truncation optimization method cut based on S1 digestion - Google Patents
A kind of aptamers truncation optimization method cut based on S1 digestion Download PDFInfo
- Publication number
- CN110511935A CN110511935A CN201910835321.6A CN201910835321A CN110511935A CN 110511935 A CN110511935 A CN 110511935A CN 201910835321 A CN201910835321 A CN 201910835321A CN 110511935 A CN110511935 A CN 110511935A
- Authority
- CN
- China
- Prior art keywords
- aptamers
- minutes
- optimization
- incubated
- truncation
- 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.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/111—General methods applicable to biologically active non-coding nucleic acids
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/115—Aptamers, i.e. nucleic acids binding a target molecule specifically and with high affinity without hybridising therewith ; Nucleic acids binding to non-nucleic acids, e.g. aptamers
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/10—Type of nucleic acid
- C12N2310/16—Aptamers
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2330/00—Production
- C12N2330/30—Production chemically synthesised
Landscapes
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Genetics & Genomics (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Microbiology (AREA)
- Plant Pathology (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biophysics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
A kind of aptamers cut based on S1 digestion are truncated optimization method and belong to medical diagnostic field.This method includes magnetic bead and albumen coupling (M&P), aptamers and M&P are incubated for (M&P&A), S1 nuclease truncates optimization, mass spectral analysis.Aptamers realize that the truncation of sequence length optimizes by the method.Based on steric hindrance, nuclease cutting aptamers are tightly combined that sector sequence efficiency is lower, which is largely saved and show biggish abundance of ions in Mass Spectrometer Method with target.Sequence and the KD value of target PD-L1 albumen demonstrate the feasibility of this method than former sequence decline 54.67% after truncation.The present invention can optimize for the truncation of aptamers provides the method based on experiment, and can be used for determining the bond area of aptamers and target.
Description
Technical field
The invention belongs to medical diagnostic field, it is related specifically to screen the side of aptamers using nuclease digestion optimization
Method.The truncation optimization of aptamers is mainly software calculating simulation at present, is seldom related to the direct truncation optimization based on experiment.The party
Method can optimize for the truncation of aptamers provide experimental method, and for illustrate the interaction mechanism of aptamers and target provide according to
According to.
Background technique
As Chinese society rapid economic development, aging of population process accelerate, cancer has become to have become and be only second to
The high mortality disease of cardiovascular disease.Cancer is chronic disease, and the early early treatment that diagnoses is for improving survival with great
Meaning.Tumor-marker analyte detection is the important means of cancer early detection, identification, prognosis, regardless of in patient or healthy person body
Tumor markers content be micro, it is therefore desirable to highly sensitive detection method.Currently, the detection method of tumor marker
Mainly detected using the specificity interaction between albumen.Programmed death ligand 1 (PD-L1) is a kind of important swells
Tumor marker, it is a kind of transmembrane protein, and the overexpression on a variety of cancer cells is apoptosis receptor 1 (PD-1)
Major ligand.PD-L1 albumen plays an important role in adjusting antitumor host immune, and tumour cell is made to escape host immune
System simultaneously promotes cancer metastasis.
The expression and purification of antibody protein is with high costs and easy in inactivation, and difference is big between batch, to tumor markers testing result
Generate adverse effect.Aptamer be from it is being screened in the library DNA or RNA, can be with the list in conjunction with target substance high specific
Chain oligonucleotide sequence.Aptamer passes through base pair stacking, hydrogen bond, electrostatic and hydrophobic effect etc. and target substance specificity
In conjunction with.Compared with protein, aptamer have affinity height, high specificity, molecular weight is small, be easily-synthesized easily modification, screen
The advantages that period is short is the good substitution of protein antibody.
The aptamer directly screened from library possibly can not directly apply to the system to be checked of complicated component,
It needs further to truncate optimization.As the length of nucleic acid increases, due to unexpectedly interacting, they are there is a greater chance that by locking
Determine into the nonactive conformation of metastable state.Nucleotide base accumulation force allows nonactive conformation and activity conformation to separate, this shows shorter
Aptamer have the larger tendency for becoming activity conformation.Aptamers truncate main by software progress molecular docking mould
It is quasi-, sequence truncation is carried out according to simulation aptamers-target action site.The present invention is using PD-L1 as target molecule, with S1 nuclease
Enzyme is truncated, is had an adverse effect based on aptamers and target bond area to digesting efficiency, the bond area piece in Mass Spectrometer Method
Section shows biggish abundance of ions, and truncates the higher sequence of optimization affinity.
Summary of the invention
It is an object of the invention to overcome the shortcomings of that existing aptamers truncate optimization, novel aptamers are provided and truncate optimization
Method has more the fact compared to software-based molecular dynamics simulation, and to illustrate phase interaction of the aptamers with target
Foundation is provided with mechanism.The present invention is to truncate enzyme with S1 nuclease, truncates to known aptamers using PD-L1 as target molecule
The higher sequence of optimization affinity.
Above-mentioned purpose is achieved through the following technical solutions:
1. synthesizing PD-L1 aptamer shown in following sequence:
5’-GCT GTG TGA CTC CTG CAA GACGGACCA GCC TTG CCG CAA GAC GGA CCA GGG
ATT CAAACG AGC AGC TGT ATC TTG TCT CC-3’。
2. truncating optimization aptamers using nucleic acid digestion
Digestion truncates optimization process agents useful for same: 2 × protein binding liquid concentration composition are as follows: 100mM sodium phosphate, 600mM
NaCl, 0.02% Tween-20, pH8.0 (2 × protein binding liquid is diluted 1 times by 1 × protein binding liquid).
SELEX combination liquid: 150mM NaCl, 5mM KCl, 1mM MgCl2, 1mM CaCl2, 40mM 4- hydroxyethyl piperazine second
Sulfonic acid, pH 7.4.
Eluent: 20mM trishydroxymethylaminomethane, 500mM imidazoles, pH 7.5.
5 × reaction solution: 200mM sodium acetate, 1.5M NaCl, 10mM ZnSO4, pH 4.5.
Human PD-L 1/B7-H1 albumen, His Tag (HPLC-verified) are bought from Acrobiosystems
(Bethesda, MD).
2.1 PD-L1 albumen are rebuild
The PD-L1 of freeze-drying is reconfigured to the stoste of 400 μ g/mL with 250 μ L aseptic deionized waters, it is solubilized at room temperature
30-60 minutes.10 parts are divided equally into, every part of 25 μ L are placed in -80 DEG C of refrigerators.
2.2 PD-L1 albumen are connected with magnetic bead
2.2.1 5 μ L PD-L1 protein solutions is taken to be added in 700 μ L 1 × protein binding liquid.
2.2.2 magnetic bead (be vortexed > 30 seconds or tilt, rotate 5 minutes) is mixed in the vial.
2.2.3 50 μ L magnetic beads are transferred on micro-centrifuge tube, and be placed on Magneto separate frame 2 minutes, aspirated and remove
Clear liquid.Sample (making in 1 × protein binding liquid) is added into magnetic bead, mixes.
2.2.4 it is incubated for 5 minutes on roller at 37 DEG C, being incubated for terminates to be placed in Magneto separate frame 2 minutes, abandons supernatant.
2.2.5 it takes 1 × protein binding liquid to wash magnetic bead-protein complexes 3 times (300 μ L every time), abandons within Magneto separate 2 minutes
Clear liquid.
2.3 aptamers are incubated for
2.3.1 aptamers are dissolved in sterile water, 95 DEG C thermal denaturation 5 minutes, ice bath 10 minutes, the liquid in conjunction with isometric SELEX
It mixes.
2.3.2 it is added in magnetic bead-protein complex, mixes, be incubated for 30 minutes at room temperature.
2.3.3 being incubated for, which terminates Magneto separate, removes supernatant, with PBS-T buffer solution for cleaning 2 times (200 μ L every time).
2.4 S1 nucleases are tested conscientiously
2.4.1 the bead complexes and appropriate S1 nuclease for taking 2.3.3 to obtain are added in 1 × reaction solution system, and 37 DEG C
Reaction 30 minutes, Magneto separate is washed 2 times (500 μ L every time) with PBS-T, removes supernatant.
2.4.2 200 μ L eluents are added, are uniformly mixed, (attention is twined 100 DEG C of boiling water baths with adhesive tape when boiling within 10 minutes
It is centrifuged nozzle, is inserted among foam or sponge and is put into boiling water), Magneto separate takes supernatant.
3. mass spectral analysis
3.1 analyze supernatant obtained by 2.4.2 with the silent winged LTQXL Linear ion trap mass spectrometer of match, as a result such as Fig. 1.
3.2 comparison mass spectrograms and truncation presequence obtain truncation optimization postorder and are classified as:
5'-AAGACGGACCAGCCTTGCCGCAAGACGGACCAGGGATT-3'.Fig. 2 is to truncate aptamer in original
The secondary structure figure of position in aptamers.
4 affinity analysis:
4.1 is suitable by PD-L1 after the former PD-L1 aptamer of 5 ' end mark fluorescent groups and truncation with SELEX combination liquid
Ligand is configured to a series of concentration respectively.
4.2 each concentration gradients take 100 μ L, 95 DEG C water-bath 5 minutes, ice bath 10 minutes, be added into PD-L1-His label
In bead complexes, 37 DEG C are incubated for 30 minutes.
4.3 clean the complex of magnetic bead-PD-L1- aptamers in 4.2 2 times (200 μ L every time) with 1 × protein binding liquid,
200 μ L are finally resuspended in, survey fluorescence intensity with microplate reader.
4.4 using aptamer concentration as abscissa, and fluorescence intensity is ordinate, draws and combines saturation curve, acquires original
PD-L1 aptamers Kd value is 115.15nM, and PD-L1 aptamers Kd value is 52.20nM after truncation.As shown in figure 3, being fitted after truncating
Ligand Kd value reduces by 54.67%, illustrates that aptamers have carried out effective truncation optimization.
Advantages of the present invention:
(1) compared with tradition truncates optimisation technique (software prediction truncation), this method has based on the straight of experiment the present invention
Connect the advantage for truncating optimization, and strong operability.
(2) present invention provides fact basis to illustrate the interaction mechanism of aptamers and target.
(3) present invention truncates the aptamer that optimization obtains, can be in conjunction with PD-L1, and affinity is more than former aptamers
It is high.
(4) the aptamer modified different report molecule is utilized in the present invention, can construct various biosensors,
For detecting the excretion body for carrying PD-L1 in human plasma, the cancer cell of the high table PD-L1 of separation detection can be used for.
Detailed description of the invention
Fig. 1: mass spectral results figure after aptamer digestion
Fig. 2: the secondary structure figure of aptamer position in former aptamers is truncated
Fig. 3: kd value nonlinear fitting (the A. original sequence of optimization front and back aptamer is truncated；B. truncated sequence)
Specific embodiment
Experimental method used in following embodiments is conventional method unless otherwise specified.
The materials, reagents and the like used in the following examples is commercially available unless otherwise specified.
Embodiment 1: PD-L1 aptamer shown in following sequence is synthesized
1.PD-L1 aptamer:
5’-GCT GTG TGA CTC CTGCAA GAC GGA CCA GCC TTG CCG CAA GAC GGA CCA GGG
ATT CAAACG AGC AGC TGT ATC TTG TCT CC-3’。
Embodiment 2: optimization aptamers are truncated using nucleic acid digestion
2. digestion truncates optimization
Digestion truncates optimization process agents useful for same: 2 × protein binding liquid final concentration composition are as follows: 100mM sodium phosphate, 600mM
NaCl, 0.02% Tween-20, pH8.0 (2 × protein binding liquid is diluted 1 times by 2 × protein binding liquid).
SELEX combination liquid: 150mM NaCl, 5mM KCl, 1mM MgCl2, 1mM CaCl2, 40mM 4- hydroxyethyl piperazine second
Sulfonic acid, pH 7.4.
Eluent: 20mM trishydroxymethylaminomethane, 500mM imidazoles, pH 7.5.
Human PD-L 1/B7-H1 albumen, His Tag (HPLC-verified) are bought from Acrobiosystems
(Bethesda, MD).
2 × Reaction Buffer:200mM sodium acetate, 1.5M NaCl, 10mM ZnSO4, pH 4.5.
2.1 PD-L1 albumen are rebuild
The PD-L1 of freeze-drying is reconfigured to the stoste of 400 μ g/mL with 250 μ L aseptic deionized waters, it is solubilized at room temperature
30-60 minutes.10 parts are divided equally into, every part of 25 μ L are placed in -80 DEG C of refrigerators.
2.2 PD-L1 albumen are connected with magnetic bead
2.2.1 5 μ L PD-L1 protein solutions is taken to be added in 700 μ L 1 × protein binding liquid.
2.2.2 magnetic bead (be vortexed > 30 seconds or tilt, rotate 5 minutes) is mixed in the vial.
2.2.3 50 μ L magnetic beads are transferred on micro-centrifuge tube, and be placed on Magneto separate frame 2 minutes, aspirated and remove
Clear liquid.Sample (making in 1 × protein binding liquid buffer) is added into magnetic bead, mixes.
2.2.4 it is incubated for 5 minutes on roller at 37 DEG C, being incubated for terminates to be placed in Magneto separate frame 2 minutes, abandons supernatant.
2.2.5 it takes 1 × protein binding liquid to wash magnetic bead-protein complexes 3 times (300 μ L every time), abandons within Magneto separate 2 minutes
Clear liquid.
2.3 aptamers are incubated for
2.3.1 aptamers are dissolved in sterile water, 95 DEG C thermal denaturation 5 minutes, ice bath 10 minutes, the liquid in conjunction with isometric SELEX
It mixes.
2.3.2 it is added in magnetic bead-protein complex, mixes, be incubated for 30 minutes at room temperature.
2.3.3 being incubated for, which terminates Magneto separate, removes supernatant, with PBS-T buffer solution for cleaning 2 times (200 μ L every time).
2.4S1 nuclease is tested conscientiously
2.4.1 the bead complexes and appropriate S1 nuclease for taking 2.3.3 to obtain are added in 1 × reaction solution system, and 37 DEG C
Reaction 30 minutes, Magneto separate is washed 2 times (500 μ L every time) with PBS-T, removes supernatant.
2.4.2 200 μ L eluents are added, are uniformly mixed, (attention is twined 100 DEG C of boiling water baths with adhesive tape when boiling within 10 minutes
It is centrifuged nozzle, is inserted among foam or sponge and is put into boiling water), Magneto separate takes supernatant.
3. mass spectral analysis of embodiment
3.1 analyze supernatant obtained by 2.4.2 with the silent winged LTQXL Linear ion trap mass spectrometer of match, as a result such as Fig. 1.
3.2 comparison mass spectrograms and truncation presequence obtain truncation optimization postorder and are classified as: 5 '-AAG ACGGAC CAG CCT
TGC CGC AAG ACG GAC CAG GGA TT-3'.Fig. 2 is the second level knot for truncating aptamer position in former aptamers
Composition.
4 affinity analysis of embodiment:
4.1 is suitable by PD-L1 after the former PD-L1 aptamer of 5 ' end mark fluorescent groups and truncation with SELEX combination liquid
Ligand is configured to a series of concentration respectively.
4.2 each concentration gradients take 100 μ L, 95 DEG C water-bath 5 minutes, ice bath 10 minutes, be added into PD-L1-His label
In bead complexes, 37 DEG C are incubated for 30 minutes.
4.3 clean the complex of magnetic bead-PD-L1- aptamers in 4.2 2 times (200 μ L every time) with 1 × protein binding liquid,
200 μ L are finally resuspended in, survey fluorescence intensity with microplate reader.
4.4 using aptamer concentration as abscissa, and fluorescence intensity is ordinate, draws and combines saturation curve, acquires original
PD-L1 aptamers Kd value is 115.15nM, and PD-L1 aptamers Kd value is 52.20nM after truncation.As shown in figure 3, being fitted after truncating
Ligand Kd value reduces by 54.67%, illustrates that aptamers have carried out effective truncation optimization.
Sequence table
<110>Beijing University of Chemical Technology
<120>a kind of aptamers cut based on S1 digestion truncate optimization method
<141> 2019-01-28
<160> 2
<170> SIPOSequenceListing 1.0
<210> 2
<211> 80
<212> DNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 2
gctgtgtgac tcctgcaaga cggaccagcc ttgccgcaag acggaccagg gattcaaacg 60
agcagctgta tcttgtctcc 80
<210> 2
<211> 38
<212> DNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 2
aagacggacc agccttgccg caagacggac cagggatt 38
Claims (2)
1. a kind of aptamers cut based on S1 digestion truncate optimization method, which is characterized in that
Using PD-L1 as target molecule, it is to truncate enzyme with S1 nuclease, is achieved through the following technical solutions:
1) PD-L1 aptamer shown in following sequence is synthesized:
5’-GCT GTG TGA CTC CTG CAA GAC GGA CCA GCC TTG CCG CAA GAC GGA CCA GGG
ATT CAA ACG AGC AGC TGT ATC TTG TCT CC-3’。
2) optimization aptamers are truncated using nucleic acid digestion, specific as follows:
Digestion truncates optimization process agents useful for same: 2 × protein binding liquid concentration composition are as follows: 100mM sodium phosphate, 600mM NaCl,
0.02% Tween-20, pH8.0；2 × protein binding liquid is diluted 1 times by 1 × protein binding liquid；
SELEX combination liquid: 150mM NaCl, 5mM KCl, 1mM MgCl2, 1mM CaCl2, 40mM 4- hydroxyethyl piperazine second sulphur
Acid, pH 7.4；
Eluent: 20mM trishydroxymethylaminomethane, 500mM imidazoles, pH 7.5；
5 × reaction solution: 200mM sodium acetate, 1.5M NaCl, 10mM ZnSO4, pH 4.5；
2.1 PD-L1 albumen is rebuild
2.2 PD-L1 albumen are connected with magnetic bead
2.2.1 5 μ L PD-L1 protein solutions is taken to be added in 700 μ L 1 × protein binding liquid；
2.2.2 mixing magnetic bead in the vial
2.2.3 50 μ L magnetic beads are transferred on micro-centrifuge tube, and be placed on Magneto separate frame 2 minutes, aspirated and remove supernatant
Liquid；Sample is added into magnetic bead, mixes；
2.2.4 it is incubated for 5 minutes on roller at 37 DEG C, being incubated for terminates to be placed in Magneto separate frame 2 minutes, abandons supernatant；
2.2.5 1 × protein binding liquid is taken to wash magnetic bead-protein complexes 3 times, 2 minutes abandoning supernatants of Magneto separate；
2.3 aptamers are incubated for
2.3.1 aptamers are dissolved in sterile water, 95 DEG C thermal denaturation 5 minutes, ice bath 10 minutes, liquid was mixed in conjunction with isometric SELEX
It is even；
2.3.2 it is added in magnetic bead-protein complex, mixes, be incubated for 30 minutes at room temperature；
2.3.3 being incubated for, which terminates Magneto separate, removes supernatant, with PBS-T buffer solution for cleaning 2 times；
2.4S1 nuclease is tested conscientiously
2.4.1 the bead complexes and S1 nuclease for taking 2.3.3 to obtain are added in 1 × reaction solution system, and 37 DEG C are reacted 30 points
Clock, Magneto separate are washed 2 times with PBS-T, remove supernatant；
2.4.2 200 μ L eluents are added, be uniformly mixed, 100 DEG C boiling water bath 10 minutes, Magneto separate takes supernatant.
2. sequence after a kind of truncation optimization, sequence are as follows:
5’-AAGACGGACCAGCCTTGCCGCAAGACGGACCAGGGATT-3’。
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2019100781507 | 2019-01-28 | ||
| CN201910078150 | 2019-01-28 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110511935A true CN110511935A (en) | 2019-11-29 |
| CN110511935B CN110511935B (en) | 2021-06-08 |
Family
ID=68631160
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910835321.6A Active CN110511935B (en) | 2019-01-28 | 2019-09-05 | Aptamer truncation optimization method based on S1 enzyme cleavage |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110511935B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111321204A (en) * | 2020-02-09 | 2020-06-23 | 北京化工大学 | Aptamer sensor-based high-sensitivity method for detecting PD-L1 in serum |
| CN113129996A (en) * | 2021-03-22 | 2021-07-16 | 复旦大学 | Aptamer optimization design method based on molecular dynamics simulation |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104630230A (en) * | 2015-01-06 | 2015-05-20 | 江南大学 | Group of nucleic acid aptamers for specifically recognizing okadaic acid |
| CN104894135A (en) * | 2015-04-28 | 2015-09-09 | 中国人民解放军第二军医大学 | High affinity adapter body capable of specifically binding with saxitoxin acetate and application thereof |
| CN108977448A (en) * | 2018-08-03 | 2018-12-11 | 湖北师范大学 | For detecting aptamer and its screening technique and the application of clenobuterol hydrochloride |
-
2019
- 2019-09-05 CN CN201910835321.6A patent/CN110511935B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104630230A (en) * | 2015-01-06 | 2015-05-20 | 江南大学 | Group of nucleic acid aptamers for specifically recognizing okadaic acid |
| CN104894135A (en) * | 2015-04-28 | 2015-09-09 | 中国人民解放军第二军医大学 | High affinity adapter body capable of specifically binding with saxitoxin acetate and application thereof |
| CN108977448A (en) * | 2018-08-03 | 2018-12-11 | 湖北师范大学 | For detecting aptamer and its screening technique and the application of clenobuterol hydrochloride |
Non-Patent Citations (5)
| Title |
|---|
| ABHIJEET DHIMAN等: "Rational truncation of aptamer for cross-species application to detect krait envenomation", 《SCI REP.》 * |
| HONGWEI HU等: "S1 nuclease digestion-based rational truncation of PD-L1 aptamer and establishment of a signal dual amplification aptasensor", 《SENSORS AND ACTUATORS: B. CHEMICAL》 * |
| WEI-YUN LAI等: "A Novel PD-L1-targeting Antagonistic DNA Aptamer With Antitumor Effects", 《MOLECULAR THERAPY—NUCLEIC ACIDS》 * |
| ZONGWEN WANG等: "Introducing structure-switching functionality into small-molecule-binding aptamers via nuclease-directed truncation", 《NUCLEIC ACIDS RES.》 * |
| 孙玉琼等: "核酸适配体序列优化策略的研究进展", 《化学传感器》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111321204A (en) * | 2020-02-09 | 2020-06-23 | 北京化工大学 | Aptamer sensor-based high-sensitivity method for detecting PD-L1 in serum |
| CN111321204B (en) * | 2020-02-09 | 2021-10-01 | 北京化工大学 | Aptamer sensor-based high-sensitivity method for detecting PD-L1 in serum |
| CN113129996A (en) * | 2021-03-22 | 2021-07-16 | 复旦大学 | Aptamer optimization design method based on molecular dynamics simulation |
| CN113129996B (en) * | 2021-03-22 | 2022-04-12 | 复旦大学 | Aptamer optimization design method based on molecular dynamics simulation |
Also Published As
| Publication number | Publication date |
|---|---|
| CN110511935B (en) | 2021-06-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Tang et al. | Application of magnetic nanoparticles in nucleic acid detection | |
| Zhang et al. | Rapid capture and nondestructive release of extracellular vesicles using aptamer-based magnetic isolation | |
| ES2617226T3 (en) | Multiplexed trials based on aptamers | |
| Wang et al. | Fabrication of an aptamer-coated liposome complex for the detection and profiling of exosomes based on terminal deoxynucleotidyl transferase-mediated signal amplification | |
| JP2021129577A (en) | Methods for generating aptamers with improved off-rates | |
| ES2424269T3 (en) | Immuno-PCR sandwich sandwich | |
| Wu et al. | Aptamer-based liquid biopsy | |
| KR20160080815A (en) | Oligonucleotide probes and uses thereof | |
| CN105452466A (en) | Aptamers and uses thereof | |
| Jin et al. | Multivalence-actuated DNA nanomachines enable bicolor exosomal phenotyping and PD-L1-guided therapy monitoring | |
| Wang et al. | Immunomagnetic antibody plus aptamer pseudo-DNA nanocatenane followed by rolling circle amplication for highly-sensitive CTC detection | |
| CN110511935A (en) | A kind of aptamers truncation optimization method cut based on S1 digestion | |
| Ahn et al. | Aptamer microarray mediated capture and mass spectrometry identification of biomarker in serum samples | |
| Nurwidya et al. | Circulating tumor cell and cell-free circulating tumor DNA in lung cancer | |
| EP3369820B1 (en) | Dna aptamer capable of bonding to vwf | |
| CN109337909A (en) | It is a kind of detect liver cancer drug-resistant cell strain aptamer and its application | |
| US20200362349A1 (en) | Application of aptamer in recognition and binding of alkaline phosphatase heterodimer or tumor detection | |
| Sun et al. | Combination of immunomagnetic separation with aptamer-mediated double rolling circle amplification for highly sensitive circulating tumor cell detection | |
| Shang et al. | Extracellular vesicles: a brief overview and its role in precision medicine | |
| CN104988154A (en) | Application of nucleic acid aptamers in identifying and being bound with integrin alpha4 | |
| WO2017094733A1 (en) | Dna aptamer binding to molecular targeting drug and method for detecting molecular targeting drug using same | |
| Feng et al. | Multi-parameter inputted logic-gating on aptamer-encoded extracellular vesicles for colorectal cancer diagnosis | |
| Xue et al. | Stimuli-induced upgrade of nuclease-resistant DNA nanostructure composed of a single molecular beacon for detecting mutant genes | |
| TW201643249A (en) | Dna aptamer capable of binding to non-small cell lung cancer cell (h1975) | |
| US20220073910A1 (en) | A Cross-Linking Approach to Map Small Molecule-RNA Binding Sites in Cells |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |