CN101236208B - Method for identifying blood albumen by chemically modified zinc oxide nanometer rod array film - Google Patents
Method for identifying blood albumen by chemically modified zinc oxide nanometer rod array film Download PDFInfo
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- CN101236208B CN101236208B CN2007100483740A CN200710048374A CN101236208B CN 101236208 B CN101236208 B CN 101236208B CN 2007100483740 A CN2007100483740 A CN 2007100483740A CN 200710048374 A CN200710048374 A CN 200710048374A CN 101236208 B CN101236208 B CN 101236208B
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Abstract
The invention discloses a serum protein identification method through chemical modified ZnO nanometer rod array thin film. The invention adopts a fluorescence spectrometer to detect the array thin film and determines the serum protein through luminous intensity of the biologically connected nanometer rod array. The invention has the following steps in order: firstly connecting a polypeptide condensing agent to the surface of chemically modified ZnO nanometer rod array thin film; then connecting the serum protein to the ZnO nanometer rod array thin film; finally analyzing the biological array thin film with the fluorescence spectrometer, and identifying the serum protein through the detected luminous intensity. The invention adopts the chemical modification method to graft the serum protein onto the surface of the ZnO nanometer rod array to form the biological array thin film, and adopts the fluorescence spectrometer to carryout biological signal detection to identify the serum protein, thereby enabling the ZnO nanometer rod to have biological detection practical significance. The invention has wide application value in the biological detection and the disease diagnosis fields, etc.
Description
Technical field
The invention relates to the method for fluoroscopic examination identification haemocyanin, especially adopt chemical modification ZnO nano-stick array thin film to discern the method for haemocyanin.
Background technology
Zinc paste (ZnO) is that zinc paste is as a kind of important semiconductor and piezoelectric nano material; Band gap width and exciton binding energy are respectively 3.37eV and 60meV; At room temperature can obtain excitonic luminescence efficiently, it is with a wide range of applications at aspects such as vacuum fluorescence demonstration, electroluminescence, low pressure FED, laser, optoelectronic device, sensor and energy converters.Compare with other semiconductor nano materials, nano zine oxide has unusual abundant special appearance, like: flower-shaped, ring-type, band shape, four needle-likes, on chip, tower shape, cage shape and propeller-like etc.These abundant special appearances possibly make it have some new performances and be expected to play a significant role at aspects such as nano-device and microelectronic devices, are considered to be only second to CNT most important nano structural material afterwards.
Because have rapidly and efficiently, in enormous quantities, characteristics such as high sensitivity, selectivity; The fluorescence of biomolecule is widely used in many analyses such as protein component and function; Connect immunosorbent detection, fluorescent colloid dyeing and protein detection like enzyme, and gene sequencing, proteomics, new drug development and medical diagnosis on disease etc.In these biologicals detected, improving detection sensitivity, increasing signal to noise ratio (S/N ratio) was the challenge of the current maximum that faces.For improving fluorescent detection capabilities and resolution, people have developed advanced base material.A collection of new, can stop the organic and inorganic and compound substance of fluorescence molecule cancellation property to be developed, they can produce the multi-fluorescence effect under single excitation source.The development of new detection technique of fluorescence is that current nano science is to one of maximum contribution in biomolecule fluoroscopic examination field.Characteristics such as nano-ZnO has the excitation wave length and width, emission spectrum is narrow, luminous intensity is high, no photobleaching, thus in detection, have be swift in response with the good characteristics of sensitivity.The broad-band gap characteristic of ZnO nano wire is also rare to be used in biology sensor.The more important thing is, adopt the ZnO nano-stick array thin film, can firmly be connected, make this film have more Practical significance widely, can be used for that biological information detects and during medical diagnosis on disease analyzes with biomolecule such as albumen.
Summary of the invention
The objective of the invention is the biologic array of albumen grafting and modifying at nanorod surfaces, formation albumen detected with fluorospectrophotometer in order to make the ZnO nanorod surfaces of forming array film can discern haemocyanin.Proposed to use the ZnO nano-stick array thin film for detecting the method that substrate detects the identification haemocyanin first.
1. the method with chemical modification ZnO nano-stick array thin film identification haemocyanin of the present invention in turn includes the following steps:
(1) the PBS low temperature jog of the ZnO array film of chemical modification being put into the polypeptide condensation agent soaks certain hour;
(2) base material taken out and clean five times with PBS after, put that the low temperature jog soaks certain hour in the PBS of haemocyanin into;
(3) once more base material is taken out and, dry up, obtain haemocyanin grafting ZnO nano-stick array thin film with warm braw with after the PBS cleaning five times;
(4) analyze this biologic array film with fluorospectrophotometer, through excitating light strength identification haemocyanin.
The invention has the beneficial effects as follows that grafting has formed the haemocyanin biologic array on chemical modification ZnO nano-stick array thin film, this biologic array can strengthen the ZnO photoluminescence intensity; Discern the detection haemocyanin according to the difference of different albumen luminous intensities.Realize the application of ZnO nano-stick array thin film on biological information detects, had very big Practical significance.
Description of drawings
Fig. 1: the stereoscan photograph of bovine serum albumin embodiment 1# in the grafting.
Embodiment
Below in conjunction with embodiment and comparative example the present invention is done detailed elaboration, but therefore do not limit the present invention within the described scope of embodiments.
The ZnO array film of chemical modification is put in the PBS (pH is 8.0) of the EDC of 0.08M and and soaked 48h in 8 ℃ of following jogs;
After base material taken out and clean five times with PBS, put in the PBS (pH is 8.0) of the bovine serum albumin of 0.08m/gmL and and soak 48h in 8 ℃ of following jogs;
Once more base material is taken out and, dry up, obtain haemocyanin grafting ZnO nano-stick array thin film with warm braw with after the PBS cleaning five times;
Analyze this biologic array film with fluorospectrophotometer, through excitating light strength identification haemocyanin.
The present invention compares with existing detection technique of fluorescence; The key distinction is that the ZnO nano-stick array thin film has the excitation wave length and width as detecting substrate, characteristics such as emission spectrum is narrow, luminous intensity is high, no photobleaching, thereby in detection, has and be swift in response and the good characteristics of sensitivity.Practical implementation of the present invention and comparative example are seen table 1 for details.
| The polypeptide condensation agent | Soak time | Soaking temperature | Haemocyanin | Soak time | Soaking temperature | Excitation wavelength | |
| 1# | EDC | 48h | 8℃ | Bovine serum albumin | 48h | 8℃ | 340nm |
| 2# | EDC | 24h | 8℃ | Bovine serum albumin | 48h | 8℃ | 340nm |
| 3# | EDG | 48h | 8℃ | The human albumin | 48h | 8℃ | 340nm |
| 4# | EDC | 48h | 8℃ | The human serum globulin | 48h | 4℃ | 340nm |
| 5# | EDC | 48h | 1℃ | The saccharification haemocyanin | 48h | 1℃ | 340nm |
The embodiment of ZnO nano-stick array thin film and comparative example's performance test results are seen table 2.
Adopt fluorospectrophotometer to detect the excitation wavelength and the light intensity of grafted protein.
| Wavelength of transmitted light | The emission spectrum half-peak breadth | Emitted luminescence intensity | |
| 1# | 365～400nm | 16nm | 2100 |
| 2# | 368～405nm | 14nm | 1900 |
| 3# | 362～415nm | 20nm | 3800 |
| 4# | 366～400nm | 13nm | 1600 |
| 5# | 368～400nm | 12nm | 1450 |
The testing result of table 2 shows that different haemocyanins is grafted on the excitation light irradiation of ZnO nano-stick array thin film with 320nm, has obtained distinct transmit spectrum and emitted luminescence intensity, and particularly the emitted luminescence intensity difference is obviously told different haemocyanins.
Claims (1)
1. method with chemical modification ZnO nano-stick array thin film identification haemocyanin, step is following successively for it:
(1) the ZnO array film of chemical modification is immersed the low temperature jog soaks a period of time in the PBS of polypeptide condensation agent;
(2) base material taken out and clean five times with PBS after, put then that the low temperature jog soaks a period of time in the PBS of haemocyanin into;
(3) dry up once more with the base material taking-up and with after the PBS cleaning five times, and with warm braw, obtain haemocyanin grafting ZnO nano-stick array thin film;
(4) analyze this biologic array film with fluorospectrophotometer, through excitating light strength identification haemocyanin;
Wherein: the described polypeptide condensation agent of step (1) is 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride (EDC), N; N '-DIC, N; N '-dicyclohexylcarbodiimide, N-hydroxyl 5-ENB-2,3-dihydroxy imines, 1-(3-dimethylamino-propyl)-3-ethyl carbodiimide; The polypeptide condensation agent is 0.001～0.1M in concentration of phosphate buffer; The pH value of step (1) and the described PBS of step (2) is 6～9, soaking temperature is that 0～10 ℃, soak time are 2h～72h; The described haemocyanin of step (2) is bovine serum albumin, human albumin, serum ferritin, saccharification haemocyanin; Its concentration at phosphate buffer solution is 0.1～500mg/mL; Used excitation wavelength is 200～500nm in the described spectral analysis of step (4).
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| CN2007100483740A CN101236208B (en) | 2007-02-01 | 2007-02-01 | Method for identifying blood albumen by chemically modified zinc oxide nanometer rod array film |
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| CN2007100483740A CN101236208B (en) | 2007-02-01 | 2007-02-01 | Method for identifying blood albumen by chemically modified zinc oxide nanometer rod array film |
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| CN101236208A CN101236208A (en) | 2008-08-06 |
| CN101236208B true CN101236208B (en) | 2012-07-04 |
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Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102650591A (en) * | 2012-04-06 | 2012-08-29 | 上海蓝怡科技有限公司 | Kit for determining glycated serum protein |
| CN104401955B (en) * | 2014-10-24 | 2016-11-30 | 西北工业大学 | A kind of BSA/Zn3(PO4)3the preparation method of hybridized nanometer flower |
| CN105203518A (en) * | 2015-10-08 | 2015-12-30 | 三峡大学 | Fluorescent reagent and preparation method and application thereof |
| EP3488228B1 (en) | 2016-07-21 | 2021-05-26 | Temasek Polytechnic | Biomarker detection system |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1763263A (en) * | 2005-09-27 | 2006-04-26 | 清华大学 | Oriented ZnO nanorod or nanowire film and preparation process thereof |
| CN1769545A (en) * | 2004-11-02 | 2006-05-10 | 清华大学 | Method for developping directionally aligning zinc oxide nanometer rod array on silicon substrate |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1769545A (en) * | 2004-11-02 | 2006-05-10 | 清华大学 | Method for developping directionally aligning zinc oxide nanometer rod array on silicon substrate |
| CN1763263A (en) * | 2005-09-27 | 2006-04-26 | 清华大学 | Oriented ZnO nanorod or nanowire film and preparation process thereof |
Non-Patent Citations (2)
| Title |
|---|
| 亓立峰等.纳米微粒材料生物学活性研究进展.《中国兽药杂志》.2004,第38卷(第8期),25-28. * |
| 司伟,翟玉春.纳米组装体系及其研究进展.《材料导报》.2005,第19卷(第11期),89-93. * |
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