CN103336112A - Method for detecting human immunoglobulin E by adopting carbon nano tube micro-cantilever biosensor - Google Patents

Method for detecting human immunoglobulin E by adopting carbon nano tube micro-cantilever biosensor Download PDF

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Publication number
CN103336112A
CN103336112A CN201310262183XA CN201310262183A CN103336112A CN 103336112 A CN103336112 A CN 103336112A CN 201310262183X A CN201310262183X A CN 201310262183XA CN 201310262183 A CN201310262183 A CN 201310262183A CN 103336112 A CN103336112 A CN 103336112A
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CN
China
Prior art keywords
cantilever
carbon nano
micro
tube
aptamer
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Pending
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CN201310262183XA
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Chinese (zh)
Inventor
黄勇
梁晋涛
李桂银
周治德
黄国银
管明源
马龙飞
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Guilin University of Electronic Technology
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Guilin University of Electronic Technology
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Application filed by Guilin University of Electronic Technology filed Critical Guilin University of Electronic Technology
Priority to CN201310262183XA priority Critical patent/CN103336112A/en
Publication of CN103336112A publication Critical patent/CN103336112A/en
Priority to CN201410270049.9A priority patent/CN104090113B/en
Pending legal-status Critical Current

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6854—Immunoglobulins
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N5/00—Analysing materials by weighing, e.g. weighing small particles separated from a gas or liquid

Abstract

The invention discloses a detection method which is high in sensitivity, fast and accurate and can rapidly measure human immunoglobulin E (hIg E). The method is realized by constructing a carbon nano tube micro-cantilever biosensor. The biosensor comprises a bracket, a substrate material, a carbon nano tube and a pick-up circuit, wherein an aptamer layer is modified on the carbon nano tube. The method comprises the following steps of: manufacturing a detection probe containing a hIg E aptamer on a carbon nano tube micro-cantilever, putting the detection probe into a sample to be detected during detection, forming a compound between the hIg E in the sample to be detected and the aptamer on the detection probe through a specific reaction and attaching the compound to the micro-cantilever; and detecting the hIg E by utilizing change relationship of deflection displacement or resonant frequency of the micro-cantilever caused by mass change of the compound generated on the micro-cantilever and positive correlation between the mass size of the compound and the concentration of the hIg E in the sample to be detected.

Description

Detect the method for human immunoglobulin(HIg) E with carbon nano-tube micro-cantilever biology sensor
Technical field
The present invention relates to biomedical engineering field, relate in particular to a kind of method that detects hIgE with the micro-cantilever biology sensor.
Technical background
Human immunoglobulin(HIg) E(Human immunoglobulin E, h IgE) be a kind of common tumor markers, h IgE detection method is a lot of at present, mainly contains radio immunoassay, enzyme-linked immunosorbent assay, fluoroimmunoassay etc.Wherein radio immunoassay has radiation, and is dangerous; Other detection method complicated operations, sensitivity is not high, is difficult to realize diagnosis and the research of early stage disease, and can't satisfy the demand of fast detecting.Need set up a kind of quick, sensitive, easy and simple to handle h IgE detection method.
Summary of the invention
Technical matters to be solved by this invention provides a kind of highly sensitive, simple in structure, quick and precisely, can realize measuring fast the detection method of h IgE.
In order to solve the problems of the technologies described above, the present invention realizes the detection of h IgE by making up 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, and carbon nano tube growth is on base material, and pick-up circuit also is modified with one deck aptamer on carbon nano-tube below base material.
The preparation process of the carbon nano-tube micro-cantilever biology sensor that the present invention uses is as follows:
1, the manufacturing of micro cantilever structure
Micro-cantilever is to be base material with Semiconducting Silicon Materials, is processed into micro cantilever structure.
2, the making of pick-up circuit
Pick-up circuit is to utilize microelectronic technique to make the silicon voltage dependent resistor (VDR) at the base material lower surface, and four voltage dependent resistor (VDR)s are connected into wheatstone bridge form.
3, semi-girder growth and coating carbon nano-tube technology
Upper surface to the base material in the abovementioned steps carries out cleaning treatment, carries out ultrasound wave with acetone, absolute ethyl alcohol, deionized water respectively and cleans, and uses Low Pressure Chemical Vapor Deposition (LPCVD) carbon nano-tube then.Also can use pyrolysismethod, cladding process or additive method carbon nano-tube coating on silica-based.
4, the modification of aptamer on the carbon nano-tube micro-cantilever
Aptamer is modified on the carbon nano-tube by hydrophobic effect, forms the detector probe of a kind of energy specific recognition hIgE, finish carbon nano-tube micro-cantilever biology sensor thereby make up; H IgE aptamer also can be to be modified on the carbon nano-tube by π-π superposition.
The present invention is as follows to the step that h IgE detects:
(1) on the carbon nano-tube micro-cantilever, makes the detector probe that contains h IgE aptamer earlier;
(2) detector probe is put into sample to be tested, hIgE forms compound by the aptamer on specific reaction and the detector probe and is attached on the micro-cantilever in the sample to be tested.
(3) concentration of h IgE is proportionate in the quality of formed compound size and the sample to be tested.
(4) mass change that produces at micro-cantilever of described compound causes micro-cantilever deflection displacement or change of resonance frequency, thereby realizes the detection to h IgE.
Description of drawings
Fig. 1 detects the carbon nano-tube micro-cantilever biology sensor synoptic diagram that h IgE uses.
Embodiment
Fig. 1 is the synoptic diagram that h IgE detects the carbon nano-tube micro-cantilever biology sensor of usefulness, 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 and constitutes micro cantilever structure, carbon nano-tube 3 be grown in base material 2 above, pick-up circuit 4 also is modified with one deck aptamer 5 on carbon nano-tube 3 below base material 2.
At first, modify the aptamer 5 that h IgE is had specific recognition in carbon nano-tube 3, form a kind of detector probe; Aptamer 5 can be to be modified on the carbon nano-tube 3 by hydrophobic effect, also can be modified on the carbon nano-tube 3 by π-π superposition.
Then, detector probe is put into sample to be tested, and the aptamer 5 in the probe reacts with the h IgE generation specific recognition in the sample, forms compound, this compound produces mass effect at the micro-cantilever biology sensor, utilizes this mass effect to realize detection to h IgE.
Embodiment 1
The step that the present invention detects h IgE is as follows:
(1) the carbon nano-tube micro-cantilever is placed contains the solution that h IgE is had the aptamer of specific recognition effect, method by ultrasonic processing, with aptamer modified on carbon nano-tube, its method of modifying can be that aptamer is modified on the carbon nano-tube by hydrophobic effect, also can be to be modified on the carbon nano-tube by π-π superposition, form a kind of detector probe that includes h IgE aptamer;
(2) sample to be tested is added drop-wise on the carbon nano-tube micro-cantilever that is modified with aptamer, at room temperature hatched 15 minutes, make aptamer and the h IgE in the sample to be tested on the bio-sensing interface that the specific recognition reaction take place, form compound;
(3) concentration of h IgE is proportionate in the quality of formed compound size and the sample to be tested.
(4) compound of Xing Chenging produces mass effect at micro-cantilever, utilizes this mass effect to realize detection to h IgE.
This experiment sampling 10 mcg/ml, 1 mcg/ml, 0.5 mcg/ml, it is respectively 47Hz, 4.2 Hz, 2.3 Hz that the compound that forms produces mass effect at the silicon micro-cantilever, and the testing result of h IgE is respectively 10.4 mcg/ml, 0.93 mcg/ml gram, 0.51 mcg/ml.

Claims (4)

1. detect the method for human immunoglobulin(HIg) E with carbon nano-tube micro-cantilever biology sensor, it detects to adopt carbon nano-tube micro-cantilever biology sensor, it is characterized in that: comprise the steps
(1) on the carbon nano-tube micro-cantilever, makes the detector probe that contains h IgE aptamer earlier;
(2) detector probe is put into sample to be tested, hIgE forms compound by the aptamer on specific reaction and the detector probe and is attached on the micro-cantilever in the sample to be tested;
(3) concentration of h IgE is proportionate in the quality of formed compound size and the sample to be tested;
(4) mass change that produces at micro-cantilever of described compound causes micro-cantilever deflection displacement or change of resonance frequency, thereby realizes the detection to h IgE.
2. according to the said method of claim 1, 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 constitutes micro cantilever structure, carbon nano-tube (3) be grown in base material (2) above, pick-up circuit (4) is below base material (2); It is characterized in that: on carbon nano-tube (3), also be modified with one deck aptamer (5); The method of described modification is that aptamer passes through hydrophobic effect.
3. according to the said method of claim 2, it is characterized in that: the method for described modification is by π-π superposition.
4. according to the said method of claim 2, it is characterized in that: the method for described growth is Low Pressure Chemical Vapor Deposition.
CN201310262183XA 2013-06-27 2013-06-27 Method for detecting human immunoglobulin E by adopting carbon nano tube micro-cantilever biosensor Pending CN103336112A (en)

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CN201410270049.9A CN104090113B (en) 2013-06-27 2014-06-18 A kind of concentration is the detection method of the IgE of 0.5-10 mcg/ml

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Publication number Priority date Publication date Assignee Title
CN104090113A (en) * 2013-06-27 2014-10-08 桂林电子科技大学 Method for detecting human immune globulin E with concentration of 0.5-10[mu]g/mL
CN105137062A (en) * 2015-06-03 2015-12-09 章丘维他力医疗器械有限公司 Immunoglobulin E immunoturbidimetry detection kit

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CN105115945B (en) * 2015-06-26 2018-01-19 安徽师范大学 The detection method of gamma Globulin
CN114348437B (en) * 2021-12-16 2024-04-19 管明 Geological information data acquisition device for shockproof disaster reduction

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CN103336112A (en) * 2013-06-27 2013-10-02 桂林电子科技大学 Method for detecting human immunoglobulin E by adopting carbon nano tube micro-cantilever biosensor

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Publication number Priority date Publication date Assignee Title
CN104090113A (en) * 2013-06-27 2014-10-08 桂林电子科技大学 Method for detecting human immune globulin E with concentration of 0.5-10[mu]g/mL
CN104090113B (en) * 2013-06-27 2016-08-24 桂林电子科技大学 A kind of concentration is the detection method of the IgE of 0.5-10 mcg/ml
CN105137062A (en) * 2015-06-03 2015-12-09 章丘维他力医疗器械有限公司 Immunoglobulin E immunoturbidimetry detection kit

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Application publication date: 20131002