CN104089842B - It is a kind of that to detect platelet-derivedization growth factor concentration with CNT micro-cantilever biology sensor be the method for 0.5-10 ug/ml - Google Patents
It is a kind of that to detect platelet-derivedization growth factor concentration with CNT micro-cantilever biology sensor be the method for 0.5-10 ug/ml Download PDFInfo
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- CN104089842B CN104089842B CN201410270034.2A CN201410270034A CN104089842B CN 104089842 B CN104089842 B CN 104089842B CN 201410270034 A CN201410270034 A CN 201410270034A CN 104089842 B CN104089842 B CN 104089842B
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Abstract
Detecting platelet-derivedization growth factor concentration with CNT micro-cantilever biology sensor is a method of 0.5-10 μ g/mL, realizes by building a kind of CNT micro-cantilever biology sensor. This biology sensor comprises support, base material, CNT, pick-up circuit, on CNT, is also modified with one deck aptamer by π-π superposition. First on CNT micro-cantilever, first make the detector probe that contains PDGF aptamer, when detection, detector probe is put into sample to be tested, and in sample to be tested, PDGF is formed compound and is attached on micro-cantilever by the aptamer in specific reaction and detector probe; The mass change of utilizing this compound to produce on micro-cantilever causes that the concentration of PDGF in the variation relation of micro-cantilever deflection displacement or resonant frequency and the quality of this compound size and sample to be tested is proportionate, thereby realizes the detection to PDGF.
Description
Technical field
The present invention relates to biomedical engineering field, relate in particular to a kind of method that detects PDGF with micro-cantilever biology sensor.
Technical background
Platelet derived growth factor (plateler-derivedgrowthfactor, PDGF), as one of angiogenesis factor, has substantial connection with the generation development of tumour, is considered to a kind of potential tumor markers with clinical meaning. PDGF detection method is a lot of at present, mainly contains enzyme-linked immunosorbent assay, immunohistochemical method etc. Its detection method complicated operation, sensitivity is not high, is difficult to realize at tumour commitment, the highly sensitive of PDGF detected, and cannot meets the demand of fast detecting. " nanomechanicalmicrocantileveropreratedinvibratinmodeswit huseofRNAaptamerasareceptoemoleculesforlabel-freedetecti onofHCVhelicase ", KyoSeonHwangetal., BiosensorsandBioelectrontics, the 23rd volume 459-465 page, 20070602, a kind of method that detects HCV unwindase with aptamer as acceptor molecule by the vibration mode of micro-cantilever is disclosed. Application for a patent for invention CN101935008A discloses a kind of method of utilizing the micro-cantilever beam sensor that functionalized carbon nano-tube is sensitive material. PDGF normal concentration is 0.5-10 μ g/mL, need to set up at present the detection method quick, sensitive, easy and simple to handle that a kind of PDGF concentration is 0.5-10 μ g/mL.
Summary of the invention
Technical problem to be solved by this invention is to provide one quick and precisely, and PDGF concentration detection range is the assay method of the PDGF of 0.5-10 μ g/mL.
In order to solve the problems of the technologies described above, the present invention realizes the detection of PDGF by building a kind of CNT micro-cantilever biology sensor. This CNT micro-cantilever biology sensor comprises support, base material, CNT, pick-up circuit; Wherein base material is fixed on support one side formation micro cantilever structure, adopt Low Pressure Chemical Vapor Deposition (LPCVD), by carbon nano tube growth on base material, pick-up circuit, below base material, is modified with one deck aptamer by π-π superposition on CNT.
The preparation process of the CNT micro-cantilever biology sensor that the present invention uses is as follows:
1, the manufacture of micro cantilever structure
Micro-cantilever is to be base material 2 by Semiconducting Silicon Materials, is processed into micro cantilever structure.
2, the making of pick-up circuit 4
Pick-up circuit is to utilize microelectronic technique to make silicon piezo-resistance at base material 2 lower surfaces, and four piezo-resistances are connected into wheatstone bridge form.
3, cantilever beam growth and coating carbon nanotube technique
Upper surface to the base material 2 in abovementioned steps carries out cleaning treatment, carries out Ultrasonic Cleaning respectively by acetone, absolute ethyl alcohol, deionized water, then uses Low Pressure Chemical Vapor Deposition (LPCVD) carbon nano-tube. With LPCVD method carbon nano-tube steady quality, be difficult for displacement, distortion, be conducive to operation below aptamer is modified on CNT by π-π superposition, form stable detector probe.
4, the modification of aptamer on CNT micro-cantilever
Aptamer is modified on CNT by π-π superposition, forms the detector probe of a kind of energy specific recognition PDGF, thereby built CNT micro-cantilever biology sensor. Modify by π-π superposition, make aptamer on CNT, be not easy to run off, work with LPCVD method carbon nano-tube one, be convenient to compound that in subsequent step, PDGF and aptamer the form mass effect on micro-cantilever stable, thereby it is stable to cause that micro-cantilever resonant frequency changes, and is that 0.5-10 μ g/mL lays the first stone for realizing the detectable concentration scope of PDGF.
The step that the present invention detects PDGF is as follows:
(1) on CNT micro-cantilever, first make the detector probe that contains PDGF aptamer;
(2) detector probe is put into sample to be tested, in sample to be tested, the concentration range of PDGF is: 0.5-10 μ g/mL, and PDGF is formed compound and is attached on micro-cantilever by the aptamer in specific reaction and detector probe;
(3) in the quality size of the compound forming and sample to be tested, the concentration of PDGF is proportionate;
(4) mass change that described compound produces on micro-cantilever causes the variation of micro-cantilever deflection displacement or resonant frequency, thereby realizes the detection to PDGF.
Brief description of the drawings
Fig. 1 detects the CNT micro-cantilever biology sensor schematic diagram that PDGF uses.
Advantage and disadvantage of the present invention
The present invention utilizes CNT micro-cantilever biology sensor to detect PDGF, this sensor adopts Low Pressure Chemical Vapor Deposition (LPCVD), by carbon nano tube growth on base material, on CNT, be modified with one deck aptamer by π-π superposition, utilize aptamer and PDGF that specific recognition occurs and react formation compound, on micro-cantilever biology sensor, produce mass effect, realize the detection to PDGF by mass effect. Above-mentioned technical characterictic is mutual support, and acting in conjunction has realized in the time that PDGF concentration is 0.5-10 μ g/mL and quick and precisely having detected, highly sensitive, easy and simple to handle.
Detailed description of the invention
Fig. 1 is the schematic diagram that PDGF detects the CNT micro-cantilever biology sensor of use, comprises support 1, base material 2, CNT 3 and pick-up circuit 4. Wherein base material 2 is fixed on support 1 one sides formation micro cantilever structures; Upper surface at base material 2 carries out Ultrasonic Cleaning by acetone, absolute ethyl alcohol, deionized water respectively, then uses Low Pressure Chemical Vapor Deposition (LPCVD) carbon nano-tube 3; Pick-up circuit 4 is below base material 2; On CNT 3, be also modified with one deck aptamer 5 by π-π superposition.
First, on CNT 3, modify by π-π superposition the aptamer 5 that PDGF is had to specific recognition, form a kind of detector probe.
Then, detector probe is put into sample to be tested, and there is specific recognition with the PDGF in sample and react in the aptamer 5 in probe, forms compound, this compound produces mass effect on micro-cantilever biology sensor, utilizes this mass effect to realize the detection to PDGF.
Embodiment 1
The step that the present invention detects PDGF is as follows:
(1) CNT micro-cantilever is placed in and contains the solution that PDGF is had to the aptamer of specific recognition effect, by the method for ultrasonic processing, aptamer is modified on CNT by π-π superposition, forms a kind of detector probe of the PDGF of including aptamer;
(2) when the concentration of sample to be tested is 0.5-10 μ g/mL, sample to be tested is added drop-wise on the CNT micro-cantilever that is modified with aptamer, at room temperature hatch 15 minutes, aptamer on bio-sensing interface is reacted with the PDGF generation specific recognition in sample to be tested, form compound;
(3) in the quality size of the compound forming and sample to be tested, the concentration of PDGF is proportionate.
(4) compound forming produces mass effect on micro-cantilever, utilizes this mass effect to realize the detection to PDGF.
This experiment sampling 10 ug/ml, 1 ug/ml, the each 1mL of 0.5 ug/ml sample, it is respectively 39.8Hz, 4.1Hz, 1.9Hz that the compound forming produces mass effect on silicon micro-cantilever, and the testing result of PDGF is respectively 9.95 ug/ml, 1.03 ug/ml, 0.48 ug/ml.
Claims (1)
1. be a method for platelet-derivedization of 0.5-10 μ g/mL growth factor with CNT micro-cantilever biology sensor detectable concentration, it is characterized in that: it detects to adopt CNT micro-cantilever biology sensor, comprises the steps
(1) aptamer is modified on CNT by hydrophobic effect, forms a kind of detector probe of platelet-derivedization of energy specific recognition growth factor;
(2) in sample to be tested, the concentration of platelet-derivedization growth factor is 0.5-10 μ g/mL, detector probe is put into sample to be tested, and in sample to be tested, platelet-derivedization growth factor is formed compound and is attached on micro-cantilever by the aptamer in specific reaction and detector probe;
(3) in the quality size of the compound forming and sample to be tested, the concentration of platelet-derivedization growth factor is proportionate;
(4) mass change that described compound produces on micro-cantilever causes the variation of micro-cantilever deflection displacement or resonant frequency, thereby realizes the detection to platelet-derivedization growth factor;
Described CNT micro-cantilever biology sensor comprises support (1), base material (2), CNT (3), pick-up circuit (4); Described base material (2) is fixed on support (1) one side and forms micro cantilever structure, CNT (3) by Low Pressure Chemical Vapor Deposition be grown in base material (2) above, pick-up circuit (4) is below base material (2); On CNT (3), be modified with one deck aptamer (5) by π-π superposition.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410270034.2A CN104089842B (en) | 2013-06-27 | 2014-06-18 | It is a kind of that to detect platelet-derivedization growth factor concentration with CNT micro-cantilever biology sensor be the method for 0.5-10 ug/ml |
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| Application Number | Priority Date | Filing Date | Title |
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| CN2013102621651 | 2013-06-27 | ||
| CN2013102621651A CN103293294A (en) | 2013-06-27 | 2013-06-27 | Method for detecting blood platelet derivatization growth factors by using carbon nano-tube micro-cantilever biosensor |
| CN201310262165.1 | 2013-06-27 | ||
| CN201410270034.2A CN104089842B (en) | 2013-06-27 | 2014-06-18 | It is a kind of that to detect platelet-derivedization growth factor concentration with CNT micro-cantilever biology sensor be the method for 0.5-10 ug/ml |
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| Publication Number | Publication Date |
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| CN104089842A CN104089842A (en) | 2014-10-08 |
| CN104089842B true CN104089842B (en) | 2016-05-11 |
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| CN2013102621651A Pending CN103293294A (en) | 2013-06-27 | 2013-06-27 | Method for detecting blood platelet derivatization growth factors by using carbon nano-tube micro-cantilever biosensor |
| CN201410270034.2A Active CN104089842B (en) | 2013-06-27 | 2014-06-18 | It is a kind of that to detect platelet-derivedization growth factor concentration with CNT micro-cantilever biology sensor be the method for 0.5-10 ug/ml |
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| CN106770553B (en) * | 2016-12-20 | 2018-11-30 | 中国科学院苏州生物医学工程技术研究所 | The detection method of platelet derived growth factor |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1978315A (en) * | 2005-12-09 | 2007-06-13 | 清华大学 | Method for preparing carbon nano tube array |
| CN101935008A (en) * | 2010-07-30 | 2011-01-05 | 中国科学院上海微系统与信息技术研究所 | Method of micro cantilever beam sensor using functional carbon nano tubes as sensitive materials |
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| US20080160638A1 (en) * | 2006-12-27 | 2008-07-03 | David Lederman | Functionalized Microcantilever Sensor and Associated Method For Detection of Targeted Analytes |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1978315A (en) * | 2005-12-09 | 2007-06-13 | 清华大学 | Method for preparing carbon nano tube array |
| CN101935008A (en) * | 2010-07-30 | 2011-01-05 | 中国科学院上海微系统与信息技术研究所 | Method of micro cantilever beam sensor using functional carbon nano tubes as sensitive materials |
Non-Patent Citations (1)
| Title |
|---|
| nanomechanical microcantilever oprerated in vibratin modes with use of RNA aptamer as a receptor molecules for label-free detection of HCV helicase;Kyo Seon Hwang et al.;《Biosensors and Bioelectronics》;20070630;第23卷;第459-465页 * |
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| CN104089842A (en) | 2014-10-08 |
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