CN104237181A - Detection method for interaction of proteins and quantum dots in capillary tube - Google Patents
Detection method for interaction of proteins and quantum dots in capillary tube Download PDFInfo
- Publication number
- CN104237181A CN104237181A CN201410444544.7A CN201410444544A CN104237181A CN 104237181 A CN104237181 A CN 104237181A CN 201410444544 A CN201410444544 A CN 201410444544A CN 104237181 A CN104237181 A CN 104237181A
- Authority
- CN
- China
- Prior art keywords
- albumen
- quantum dot
- detection method
- kapillary
- qds
- 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.)
- Pending
Links
Abstract
The invention discloses a detection method for the interaction of proteins and quantum dots in a capillary tube, belonging to the technical field of nano-biology. The detection method is characterized in that the proteins and the quantum dots are coupled in the capillary tube and are detected by virtue of fluorescence. The detection method is more sensitive and clearer compared with the external coupling of the capillary tube. The method is rapid, accurate and simple to operate, the application of the capillary electrophoresis in the biological analysis can be enlarged, and reference can be provided for the further application of a quantum-dot biological probe.
Description
Technical field
The present invention relates to field of nano biotechnology, be specifically related to the interactional detection method of albumen-quantum dot in a kind of kapillary.
Background technology
Quantum dot (QDs), as a kind of novel fluorescent material, has that exciting light spectrum width, emission spectrum are narrow and symmetrical, Color tunable, anti-light bleaching and an advantage such as fluorescence lifetime is long.These superior performances make QDs be widely used in cell imaging and biomarker, become very important a kind of probe tool in cell imaging research gradually.
At present, the most widely used fat-soluble CdSe/ZnS quantum dot being metal organic solvent method and synthesizing in biomarker.Therefore, by methods such as hydrophilic ligand exchange, the encapsulation of amphipathic part, silane coatings, fat-soluble quantum dot is converted to water-soluble quantum dot.Water-soluble quantum dot can prepare quantum dot fluorescence probe with albumen by the method for covalent coupling or Electrostatic Absorption.
On the other hand, the micro separation technique that Capillary Electrophoresis consumes as a kind of high-resolution, highly sensitive, high-speed, high flux and low sample, has broad application prospects at field of bioanalysis.It is a kind of strong, high-resolution method (Jianhao Wang, et al.J Nanopart Res, 2013,15,1914) for detection QDs and albumen coupling.Capillary Electrophoresis can be separated QDs and bioconjugate well.At present, the detection method of Capillary Electrophoresis mainly contains UV detect, Electrochemical Detection, chemiluminescence detection, Mass Spectrometer Method and fluoroscopic examination.Wherein, the detection limit 3 number magnitude less of UV detect of fluorescence detection are very sensitive to the research of QDs and protein-interacting.Thus, we study the interaction of albumen-quantum dot in kapillary with fluorescent capillary electrophoresis tube.Albumen and QDs interact in kapillary, are separated, and detect, rapid, the robotization that achieving interacts detects, microminiaturization.
Summary of the invention
The technical problem to be solved in the present invention is: in order to study the problem of albumen and the trickleer dynamic variation of QDs, improves its application in biological field.For solving the problems of the technologies described above, the invention provides the interactional detection method of albumen-quantum dot in a kind of kapillary, the method is sensitiveer than detecting after extracapillary coupling, and it is more clear to change, and can observe out the subtle change that albumen is combined with QDs more accurately.
The technical solution adopted for the present invention to solve the technical problems is: the interactional detection method of albumen-quantum dot in a kind of kapillary, is characterized in that: by albumen and QDs coupling in kapillary, to be separated and it changes by fluoroscopic examination.
The sample that albumen of the present invention and quantum dot Loading sequence answer advanced electrophoretic velocity slow, then enter the fast sample of electrophoretic velocity, the mistiming of sample introduction should control interact in effective length at two samples, and both sample injection times are consistent with sample size.
Quantum dot described in the present invention includes but not limited to the quantum dot containing Cd, as CdSe, CdTe, CdS or CdSe/ZnS.
Albumen described in the present invention includes but not limited to use reductive agent NaBH
4the albumen of sex change, as denatured bovine serum protein (dBSA), sex change ovalbumin or sex change transferrins.
After adopting above-mentioned technical scheme, the beneficial effect that the present invention obtains is, the interactional detection method of albumen-quantum dot in kapillary provided by the invention, simple to operate, repeatable high, albumen can be observed out accurately and QDs interacts, more accurate than detecting after extracapillary coupling, sensitiveer, more clear, expand quantum dot further as the application of namo fluorescence probe in biomarker field.
Accompanying drawing explanation
Below in conjunction with drawings and Examples, the present invention is further described.
Fig. 1 is dBSA and QDs (mol ratio is 32:1) interactional electrophoretogram in kapillary: the sampling interval time is respectively (a) 0s (b) 10s (c) 20s (d) 30s (e) 40s (f) 50s (g) 60s.
Fig. 2 is interaction S in kapillary
qDs/ S
totalratio is with the change of sampling interval time.S
qDsthe integral area at QDs electrophoresis peak, S
totalit is the electrophoresis peak integral area of QDs and compound.
Fig. 3 is ovalbumin and QDs (mol ratio is 32:1) interactional electrophoretogram in kapillary: the sampling interval time is respectively (a) 0s (b) 10s (c) 20s (d) 40s.
Embodiment
Embodiment
The present invention will be described further with regard to following examples, but it is to be understood that these embodiments are only the use illustrated, and should not be interpreted as restriction of the invention process.
Embodiment 1
Interact in dBSA and QDs (mol ratio is 32:1) kapillary and detect
1, fat-soluble QDs is converted to water-soluble QDs through GSH
By 18mg GSH, 5mg KOH, 250 μ L methyl alcohol mixings, get 40 μ L mixed solutions and add in the fat-soluble quantum dot of 200 μ L, vibration 30min.After vibration terminates, add 200 μ L 1 mM NaOH, namely fat-soluble quantum dot is transferred in aqueous phase.Taking-up upper strata quantum dot, adds 1mL methyl alcohol and 30 μ L NaCl solution (30mg/mL) precipitate, and is dissolved in pH 7.4 borate buffer.Repeated precipitation twice, is finally dissolved in 200 μ L pH 7.4 borate buffers.After conversion, the concentration of QDs is constant.
2, the preparation of dBSA solution
DBSA solution passes through NaBH
4react obtained with BSA.Main step is as follows: 0.33g BSA is dissolved in 100mL pH 7.4 borate buffer, then adds 0.008g NaBH while stirring
4, stirred at ambient temperature 1h, then rises to 70 DEG C until do not have gas (H
2) produce.The concentration of last dBSA solution is 5 × 10
- 5mol/L.
3, the analysis of fluorescent capillary electrophoresis tube detects
When detecting in kapillary, first sample introduction QDs (height 10cm, sample introduction 10s), sample introduction dBSA (height 10cm, sample introduction 10s) again after the different time of interval, the mol ratio of dBSA and QDs is 32:1.
When we detect in kapillary, QDs and the dBSA sampling interval time is respectively 0s, 10s, 20s, 30s, 40s, 50s, 60s, and when finding to detect in kapillary, the electrophoresis peak of QDs reduces (Fig. 1, P rapidly
1), the electrophoresis peak of compound increases (Fig. 1, P
2), the electrophoresis peak (Fig. 1) of QDs has almost been can't see when interval 30s.According to S
qDs/ S
totalratio and the relation of sampling interval time, obtain the curve in Fig. 2, we find when the sampling interval time is greater than 40s, reaction completely, QDs participate in completely react.
Embodiment 2
Ovalbumin and interior interaction of QDs (mol ratio is 32:1) kapillary are detected
Ovalbumin is through NaBH
4sex change, all the other steps are with embodiment 1.When we detect in kapillary, QDs and the dBSA sampling interval time is respectively 0s, 10s, 20s, 40s, when finding to detect in kapillary, the electrophoresis peak of QDs reduces rapidly, and the electrophoresis peak of compound increases, and can't see the electrophoresis peak (Fig. 3) of QDs when interval 40s completely.
With above-mentioned according to desirable embodiment of the present invention for enlightenment, by above-mentioned description, relevant staff in the scope not departing from this invention technological thought, can carry out various change and amendment completely.The technical scope of this invention is not limited to the content on instructions, must determine its technical scope according to right.
Claims (7)
1. albumen in kapillary--the interactional detection method of-quantum dot, is characterized in that, by albumen with quantum dot coupling in kapillary, be separated and pass through fluoroscopic examination.
2. albumen in a kind of kapillary according to claim 1--the interactional detection method of-quantum dot, is characterized in that the sample that described albumen and quantum dot Loading sequence answer advanced electrophoretic velocity slow, then enters the fast sample of electrophoretic velocity; The mistiming of albumen and quantum dot sample introduction should ensure that both can interact in effective length.
3. the interactional detection method of albumen-quantum dot in a kind of kapillary according to claim 1 and 2, it is characterized in that the time of described albumen and quantum dot sample introduction and sample size consistent.
4. the interactional detection method of albumen-quantum dot in a kind of kapillary according to claim 3, is characterized in that described quantum dot is the quantum dot containing Cd.
5. the interactional detection method of albumen-quantum dot in a kind of kapillary according to claim 4, is characterized in that the described quantum dot containing Cd is CdSe, CdTe, CdS or CdSe/ZnS.
6. the interactional detection method of albumen-quantum dot in a kind of kapillary according to claim 3, is characterized in that described albumen is for use reductive agent NaBH
4the albumen of sex change.
7. the interactional detection method of albumen-quantum dot in a kind of kapillary according to claim 6, is characterized in that described reductive agent NaBH
4the albumen of sex change is denatured bovine serum protein, sex change ovalbumin or sex change transferrins.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410444544.7A CN104237181A (en) | 2014-09-02 | 2014-09-02 | Detection method for interaction of proteins and quantum dots in capillary tube |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410444544.7A CN104237181A (en) | 2014-09-02 | 2014-09-02 | Detection method for interaction of proteins and quantum dots in capillary tube |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104237181A true CN104237181A (en) | 2014-12-24 |
Family
ID=52225688
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410444544.7A Pending CN104237181A (en) | 2014-09-02 | 2014-09-02 | Detection method for interaction of proteins and quantum dots in capillary tube |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104237181A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104764776A (en) * | 2015-03-23 | 2015-07-08 | 常州大学 | A method of detecting quantum dot-protein binding kinetics |
| CN105136760A (en) * | 2015-09-06 | 2015-12-09 | 常州大学 | Rapid detection method for interaction of quantum dots and HAT labels |
| CN105181664A (en) * | 2015-09-06 | 2015-12-23 | 常州大学 | Method for rapidly detecting combination rate of protease Jmjd6 and quantum dots |
-
2014
- 2014-09-02 CN CN201410444544.7A patent/CN104237181A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104764776A (en) * | 2015-03-23 | 2015-07-08 | 常州大学 | A method of detecting quantum dot-protein binding kinetics |
| CN105136760A (en) * | 2015-09-06 | 2015-12-09 | 常州大学 | Rapid detection method for interaction of quantum dots and HAT labels |
| CN105181664A (en) * | 2015-09-06 | 2015-12-23 | 常州大学 | Method for rapidly detecting combination rate of protease Jmjd6 and quantum dots |
| CN105136760B (en) * | 2015-09-06 | 2018-08-14 | 常州大学 | A kind of method of quick detection quantum dot and the interaction of HAT labels |
| CN105181664B (en) * | 2015-09-06 | 2018-12-04 | 常州大学 | A kind of method of quick detection protease Jmjd6 and quantum dot association rate |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zrazhevskiy et al. | Quantum dot imaging platform for single-cell molecular profiling | |
| Zrazhevskiy et al. | Multicolor multicycle molecular profiling with quantum dots for single-cell analysis | |
| CN104597019B (en) | A kind of In-situ reaction system based on carbon quantum dot/manganese dioxide nano-plates layer and its application method for detecting glutathione content | |
| US20060223197A1 (en) | Method and apparatus for the detection of biological molecules | |
| EP2960651B1 (en) | Bioanalysis device and biomolecule analyser | |
| CN102866139B (en) | Establishment method based on surface plasma reinforcing energy transferring biosensor | |
| CN106753352A (en) | A kind of fluorescent carbon quantum dot of N doping and its preparation method and application | |
| CN107253961B (en) | It is a kind of can ratio test cysteine water soluble fluorescence sensor preparation and application | |
| CN112986197A (en) | Ratiometric fluorescent probe for detecting mercury ions, fluorescent paper chip and detection method | |
| CN104237181A (en) | Detection method for interaction of proteins and quantum dots in capillary tube | |
| Huang et al. | Electrochemiluminescent sensor based on Ru (bpy) 32+-doped silica nanoprobe by incorporating a new co-reactant NBD-amine for selective detection of hydrogen sulfide | |
| Tao et al. | Microfluidic devices with simplified signal readout | |
| CN104777316B (en) | A kind of preparation method of Western blotting paper chip based on quantum dot | |
| CN113189181A (en) | Single-cell protein quantitative analysis method based on electrophoresis technology | |
| Wang et al. | Cu 2+ modulated DNA-templated silver nanoclusters as a turn-on fluorescence probe for the detection of quinolones | |
| CN109825284B (en) | Ethylenediamine-beta-cyclodextrin modified Mn doped ZnS quantum dot and preparation method and application thereof | |
| CN105928917B (en) | A kind of silver nanoclusters sensor and its preparation method and application | |
| CN102565020A (en) | Method for quantitatively detecting protein through quantum dot resonant scattering | |
| De Silva et al. | True one cell chemical analysis: a review | |
| CN104764776A (en) | A method of detecting quantum dot-protein binding kinetics | |
| Ji et al. | Supersensitive sensing based on upconversion nanoparticles through cascade photon amplification at single-particle level | |
| CN108827921B (en) | Room-temperature phosphorescence detection method for lysozyme and application | |
| CN105241942A (en) | Method of quickly detecting concentration of glutathione on the basis of capillary electrophoresis | |
| CN103555334B (en) | CdTe/ZnS core-shell quantum dots, preparation method and application of CdTe/ZnS core-shell quantum dots | |
| CN109971478A (en) | The method that the nano particle of terbium ion doping is used for fluorescence double UV check dopamine |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20141224 |
|
| WD01 | Invention patent application deemed withdrawn after publication |