CN104089842A - Method for detecting platelet-derived growth factor with concentration of 0.5-10[mu]g/mL by using carbon nanotube micro-cantilever biosensor - Google Patents
Method for detecting platelet-derived growth factor with concentration of 0.5-10[mu]g/mL by using carbon nanotube micro-cantilever biosensor Download PDFInfo
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- CN104089842A CN104089842A CN201410270034.2A CN201410270034A CN104089842A CN 104089842 A CN104089842 A CN 104089842A CN 201410270034 A CN201410270034 A CN 201410270034A CN 104089842 A CN104089842 A CN 104089842A
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- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 title claims abstract description 43
- 239000002041 carbon nanotube Substances 0.000 title claims abstract description 42
- 229910021393 carbon nanotube Inorganic materials 0.000 title claims abstract description 42
- 238000000034 method Methods 0.000 title claims abstract description 13
- 102000010780 Platelet-Derived Growth Factor Human genes 0.000 title abstract description 3
- 108010038512 Platelet-Derived Growth Factor Proteins 0.000 title abstract description 3
- 239000000523 sample Substances 0.000 claims abstract description 32
- 239000000463 material Substances 0.000 claims abstract description 18
- 150000001875 compounds Chemical class 0.000 claims abstract description 16
- 238000001514 detection method Methods 0.000 claims abstract description 14
- 238000006073 displacement reaction Methods 0.000 claims abstract description 4
- 238000006243 chemical reaction Methods 0.000 claims abstract description 3
- 108091023037 Aptamer Proteins 0.000 claims description 22
- 238000004518 low pressure chemical vapour deposition Methods 0.000 claims description 11
- 239000003102 growth factor Substances 0.000 claims description 8
- 230000002209 hydrophobic effect Effects 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 10
- 108091008104 nucleic acid aptamers Proteins 0.000 abstract 3
- 239000000758 substrate Substances 0.000 abstract 1
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 4
- 206010028980 Neoplasm Diseases 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 2
- 239000008367 deionised water Substances 0.000 description 2
- 229910021641 deionized water Inorganic materials 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 229910052710 silicon Inorganic materials 0.000 description 2
- 239000010703 silicon Substances 0.000 description 2
- 238000004506 ultrasonic cleaning Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 1
- 108060004795 Methyltransferase Proteins 0.000 description 1
- 108091008103 RNA aptamers Proteins 0.000 description 1
- 239000002870 angiogenesis inducing agent Substances 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000012151 immunohistochemical method Methods 0.000 description 1
- 238000011898 label-free detection Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000004377 microelectronic Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 239000002210 silicon-based material Substances 0.000 description 1
- 239000004575 stone Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
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Abstract
A method for detecting a platelet-derived growth factor with the concentration of 0.5-10[mu]g/mL is realized by constructing a carbon nanotube micro-cantilever biosensor. The biosensor comprises a support, a substrate material, a carbon nanotube and a pick-up circuit, and the carbon nanotube is modified with a layer of nucleic acid aptamers through a phi-phi superposing effect. The method comprises the following steps: a detection probe containing a PDGF nucleic acid aptamer is made on a carbon nanotube micro-cantilever, the detection probe is put in a sample to be detected during detection, and PDGF in the sample to be detected and the nucleic acid aptamer on the detection probe form a compound and are attached to the micro-cantilever through a specific reaction; and a micro-cantilever deflection displacement or resonant frequency change caused by the mass change of the compound on the micro-cantilever is positively related to the concentration of PDGF in the sample to be detected in order to realize PDGF detection.
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-derived growth factor, 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." nanomechanical microcantilever oprerated in vibratin modes with use of RNA aptamer as a receptoe molecules for label-free detection of HCV helicase ", Kyo Seon Hwang et al., Biosensors and Bioelectrontics, 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 matters to be solved by this invention is to provide one quick and precisely, and PDGF concentration sensing 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 carbon nano-tube micro-cantilever biology sensor.This carbon nano-tube micro-cantilever biology sensor comprises support, base material, carbon nano-tube, 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 carbon nano-tube.
The preparation process of the carbon nano-tube 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 voltage dependent resistor (VDR) at base material 2 lower surfaces, and four voltage dependent resistor (VDR)s are connected into wheatstone bridge form.
3, semi-girder 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 with 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 in carbon nano-tube by π-π superposition, form stable detector probe.
4, the modification of aptamer on carbon nano-tube micro-cantilever
Aptamer is modified in carbon nano-tube by π-π superposition, forms the detector probe of a kind of energy specific recognition PDGF, thereby built carbon nano-tube micro-cantilever biology sensor.Modify by π-π superposition, make aptamer in carbon nano-tube, 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 resonance 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 carbon nano-tube 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 resonance frequency, thereby realizes the detection to PDGF.
Brief description of the drawings
Fig. 1 detects the carbon nano-tube micro-cantilever biology sensor schematic diagram that PDGF uses.
Advantage and disadvantage of the present invention
The present invention utilizes carbon nano-tube 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 carbon nano-tube, 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.
Embodiment
Fig. 1 is the schematic diagram that PDGF detects the carbon nano-tube micro-cantilever biology sensor of use, comprises support 1, base material 2, carbon nano-tube 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 with 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 carbon nano-tube 3, be also modified with one deck aptamer 5 by π-π superposition.
First, in carbon nano-tube 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) carbon nano-tube 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 in carbon nano-tube 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 carbon nano-tube 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, 4.1 Hz, 1.9 Hz 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 carbon nano-tube micro-cantilever biology sensor detectable concentration, it is characterized in that: it detects to adopt carbon nano-tube micro-cantilever biology sensor, comprises the steps
(1) aptamer is modified in carbon nano-tube 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 resonance frequency, thereby realizes the detection to platelet-derivedization growth factor;
Described carbon nano-tube micro-cantilever biology sensor comprises support (1), base material (2), carbon nano-tube (3), pick-up circuit (4); Described base material (2) is fixed on support (1) one side and forms micro cantilever structure, carbon nano-tube (3) by Low Pressure Chemical Vapor Deposition be grown in base material (2) above, pick-up circuit (4) is below base material (2); On carbon nano-tube (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 |
|---|---|---|---|
| CN201310262165.1 | 2013-06-27 | ||
| 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 |
| 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 true CN104089842A (en) | 2014-10-08 |
| CN104089842B 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 Expired - Fee Related 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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| 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 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106770553A (en) * | 2016-12-20 | 2017-05-31 | 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 (3)
| 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 |
| US20080160638A1 (en) * | 2006-12-27 | 2008-07-03 | David Lederman | Functionalized Microcantilever Sensor and Associated Method For Detection of Targeted Analytes |
| 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 |
-
2013
- 2013-06-27 CN CN2013102621651A patent/CN103293294A/en active Pending
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- 2014-06-18 CN CN201410270034.2A patent/CN104089842B/en not_active Expired - Fee Related
Patent Citations (3)
| 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 |
| US20080160638A1 (en) * | 2006-12-27 | 2008-07-03 | David Lederman | Functionalized Microcantilever Sensor and Associated Method For Detection of Targeted Analytes |
| 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 (2)
| Title |
|---|
| KYO SEON HWANG ET AL.: "nanomechanical microcantilever oprerated in vibratin modes with use of RNA aptamer as a receptor molecules for label-free detection of HCV helicase", 《BIOSENSORS AND BIOELECTRONICS》 * |
| 贾光 等: "《纳米碳管生物效应与安全应用》", 30 April 2010 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106770553A (en) * | 2016-12-20 | 2017-05-31 | 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 |
| 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 |
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| Publication number | Publication date |
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| CN104089842B (en) | 2016-05-11 |
| CN103293294A (en) | 2013-09-11 |
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