CN104880558A - InP-based HEMT tumor marker sensor and manufacturing method thereof - Google Patents
InP-based HEMT tumor marker sensor and manufacturing method thereof Download PDFInfo
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- CN104880558A CN104880558A CN201410073112.XA CN201410073112A CN104880558A CN 104880558 A CN104880558 A CN 104880558A CN 201410073112 A CN201410073112 A CN 201410073112A CN 104880558 A CN104880558 A CN 104880558A
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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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
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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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
Abstract
The invention discloses an InP-based HEMT tumor marker sensor and a manufacturing method thereof. The InP-based HEMT tumor marker sensor comprises an InP-based HEMT energy transducer, a biomolecule carrier and a biosensitive membrane, the energy transducer comprises a source electrode, a drain electrode and a grid electrode, the biomolecule carrier is formed on the grid electrode of the energy transducer, the biosensitive membrane is attached to the biomolecule carrier and can specifically identifies tumor markers, and the distribution of charges on the surface of the grid electrode changes due to influences of charges of the tumor markers, so the concentration of two dimensional electron gases in the channel of the energy transducer is influenced, thereby the source-drain current of the energy transducer changes. The InP-based HEMT tumor marker sensor improves the detection limit of the tumor markers to a new degree, and offers opportunities for prevention and treatment of malignant tumors.
Description
Technical field
The present invention relates to semiconductor devices and biochemical field, detection tumor marker sensor particularly utilizing InP-base High Electron Mobility Transistor (HEMT) to develop and preparation method thereof.
Background technology
The display of " 2012 Chinese tumour registration annual report " data, recent two decades carrys out China's tumor incidence to be continued to increase, and the whole nation is per minute just has 6 people to be diagnosed as malignant tumour, and the probability of China resident suffers from cancer is in life 22%.Lung cancer, liver cancer, cancer of the stomach, the cancer of the esophagus, colorectal cancer, breast cancer do not threaten the life of people all the time.Early detection, early diagnosis, early treatment can improve the chances of survival of patient, offer an opportunity for curing cancer patient.
Tumor markers is that a class is at the material of patient body intensive amount far above health adult's in-vivo content, their existence and quantitative change can provide scientific basis for the type of diagnosing tumour and character, have become the important supplementary means in diagnosing tumor and clinical treatment at present.By detecting oncofetal protein (CEA), neuronspecific enolase (NSE), cytokeratin (CK), squamous cell carcinoma antigen (SCCA) can effective diagnosing, thus carries out prophylactic treatment; Can play an important role at diagnosing gastric cancer and treatment by detecting alpha-fetoprotein (AFP), oncofetal protein (CEA), CA199, CA125, CA242, ferritin, CA50 etc.; In addition by detecting tumor markers, in the early diagnosis and treatment of liver cancer, the cancer of the esophagus, breast cancer etc., important effect has also been played.
Existing tumor marker sensor detects tumor markers at present clinically and generally adopts labelling immunoassay, and their ultimate principle is identical, and the signal just finally measured according to the difference of label is different.The detection method be widely used mainly contains: radiommunoassay (RIA), EIA enzyme immunoassay (EIA), time resolved fluoro-immunoassay (TFIA) and chemiluminescence immune assay (CLIA).RIA was come out the sixties in 20th century, although along with the progress of technology, adopt solid phase test tube method, simplify step, stability improves, and its radioactive contamination can not be ignored.EIA has mark preparation simply, and the term of validity is long, the advantage of environmentally safe, and after later stage nineties introducing amplification system, detection sensitivity catches up with and surpasses RIA technology, but the detection limit of first two technology is only in mol/L magnitude.1981, chemiluminescence principle combines with immune response by Pannagli, establish chemiluminescence immunoassay (CLIA), although this method sensitivity can reach the level of putting and exempting from, but affect the many factors of testing result, less stable, and after generation chemical reaction, the luminescence of sample cannot be reproduced.Time resolved fluoro-immunoassay (TFIA) and chemiluminescence immune assay (CLIA) are all detections that the fluorescence utilizing tumor markers and sensitive membrane chemical reaction to produce realizes to sample, common fluorescent marker fluorescence lifetime is very short, and this is all the key element affecting detection sensitivity.
Summary of the invention
(1) technical matters that will solve
Technical matters to be solved by this invention is the sensitivity improving tumor markers detection, makes it reach pg/ml magnitude, improves detection speed simultaneously.
The shortcoming of existing tumor markers detection technique is that detection limit can only reach mg/ml, in addition the synthesis relative difficulty of specific marker thing, although adopt the detection sensitivity of fluorescent characteristic to increase, response speed is relatively slow.
(2) technical scheme
For solving the problems of the technologies described above, the present invention proposes a kind of InP-base HEMT tumor marker sensor, for detecting tumor markers, comprising: InP-base HEMT transducer, and it comprises source electrode, drain and gate; Biomolecule carrier, is formed at the grid of described transducer; Bio-sensitive film, be attached to described biomolecule carrier, this bio-sensitive film energy specific recognition tumor markers, be subject to the impact of the electric charge of tumor markers own, described gate surface CHARGE DISTRIBUTION is changed, and then affect two-dimensional electron gas in described transducer raceway groove, the source-drain current of transducer is changed.
According to one embodiment of the present invention, described biomolecule carrier is metal, metal nanoparticle, gold size, metal oxide nano-material etc.
According to one embodiment of the present invention, described biomolecule carrier has grid voltage element, and described grid voltage element is for applying voltage.
According to one embodiment of the present invention, area of grid area distributions is between 1000 ~ 1500 μm 2.
Meanwhile, the present invention also proposes a kind of method making InP-base HEMT tumor marker sensor, comprises the steps: step S1: preparation InP-base HEMT; Step S2: make source, drain electrode on described InP-base HEMT; Step S3: the biomolecule carrier making bio-sensitive film at the area of grid of described InP-base HEMT; Step S4: prepare bio-sensitive film on the bio-sensitive film of area of grid, this bio-sensitive film energy specific recognition tumor markers, be subject to the impact of the electric charge of tumor markers own, described gate surface CHARGE DISTRIBUTION is changed, and then affect two-dimensional electron gas in described transducer raceway groove, the source-drain current of transducer is changed.
According to one embodiment of the present invention, described biomolecule carrier is Au nano particle, described step S4 is: react by using cysteamine solution and described biomolecule carrier, form S-Au key, re-use glutaraldehyde solution, making itself and described cysteamine react bonding, by using antibody to form bio-sensitive film, blocking the remaining glutaraldehyde of passivation by using sodium borohydride solution.
According to one embodiment of the present invention, described step S4 is: by using sulfydryl butanediamine aqueous solution, RuCl
3solution and gold size are fixed corresponding antibodies at described grid and are obtained bio-sensitive film.
According to one embodiment of the present invention, described biomolecule carrier is Au nano particle, and described step S4 is: form self-assembled molecule layer by biomolecule carrier being dipped in sulfydryl butanediamine aqueous solution, then by dripping RuCl to described grid
3after solution reaction a period of time, then soak in gold size, then drip antibody-solutions to grid, carry out under room temperature reacting thus fixing this antibody.
(3) beneficial effect
The invention has the advantages that the tumor marker sensor that the advantage in conjunction with InP-base HEMT high mobility, low noise is developed, the detection limit of tumor markers can be risen to a new height, expectation can reach pg/mL even fg/mL magnitude, and the prevention for malignant tumour is cured and provided first chance.
Accompanying drawing explanation
Fig. 1 is the structural representation of an embodiment of InP-base HEMT tumour mark mark thing sensor of the present invention;
Fig. 2 is the InP-base HEMT tumour mark mark thing sensor construction schematic diagram having grid voltage element for biology sensor in the present invention;
Fig. 3 is the InP-base HEMT tumour mark mark thing sensor construction schematic diagram without grid voltage element for biology sensor in the present invention.
Embodiment
InP-base HEMT has higher electron mobility, electron drift velocity and saturation drift velocity, two-dimensional electron gas etc., and there is low noise, the better characteristic of heat conductivility, the present inventor is applied in tumor marker sensor, to greatly improving the detection sensitivity, the speed of response etc. of device.
Of the present inventionly comprise InP-base HEMT transducer, biomolecule carrier and bio-sensitive film for InP-base HEMT tumor marker sensor.Described transducer comprises source electrode, drain and gate, the carrier of described biomolecule carrier and bioactive molecule, and being formed at is the grid of described transducer, and bio-sensitive film is attached to described biomolecule carrier.Bio-sensitive film energy specific recognition tumor markers, be subject to the impact of the electric charge of tumor markers own, described gate surface CHARGE DISTRIBUTION is changed, and then affects two-dimensional electron gas in described transducer raceway groove, the source-drain current of transducer is changed.
For making the object, technical solutions and advantages of the present invention clearly understand, below in conjunction with specific embodiment, and with reference to accompanying drawing, the present invention is described in further detail.
Fig. 1 is the structural representation of an embodiment of InP-base HEMT tumor marker sensor of the present invention.As shown in Figure 1, the InP-base HEMT tumor marker sensor of this embodiment has rhythmo structure, comprises InP-base HEMT transducer 10, biomolecule carrier 20 and bio-sensitive film 30.
The transducer 10 of this embodiment comprises stacked substrate 101, cushion 102, delta doping layer 103, first separation layer 104 of a Si, channel layer 105, second separation layer 106, the delta doping layer 107 of the 2nd Si, barrier layer 108, cap layers 109, source-drain electrode 110, device protecting layer 111 and contact conductor 112 from bottom to up successively.
Specifically, described substrate 101 is InP-base substrate.The material of cushion 102 is InAlAs, and improve channel carrier concentration for reducing dislocation, growth thickness is between 500nm ~ 1 μm.The dopant material of the delta doping layer 103 of the one Si is InAlAs, and for providing channel carrier, growth thickness is 0 ~ 4nm.The material of the first separation layer 104 is InAlAs, and for providing channel carrier, growth thickness is 0 ~ 4nm.The material of channel layer 105 is InGaAs, for active channel, and growth thickness 10 ~ 20nm.The material of the second separation layer 106 is InAlAs, for channel carrier and doped layer spatially being isolated, improves channel carrier mobility, growth thickness 0 ~ 4nm.The dopant material of the delta doping layer 107 of the 2nd Si is InAlAs, and for providing channel carrier, growth thickness is 0 ~ 4nm.The material of barrier layer 108 is InAlAs, and for providing the dielectric substance between Schottky contacts and two-dimensional electron gas, growth thickness is 10 ~ 20nm.The material of cap layers 109 is heavy doping InGaAs, and for providing Ohmic contact to form low-resistivity electrode, growth thickness is 10 ~ 30nm.
Source-drain electrode 110 is two, is source electrode and drain electrode, is germanium gold nickel alloy, with transducer Ohmic contact.The material of device protecting layer 111 is photoresists, for passivation and the protection of device, prevents aqueous solution etc. from having influence on units test performance.Contact conductor 112 is two, and material is Au, is connected to source-drain electrode 110, for being connected with external circuit, is convenient to sensed current signal change.
The area of grid of described biomolecule carrier 20 on the active area of described transducer 10 between source-drain electrode electrode 110, area of grid area distributions is at 1000 ~ 1500 μm
2between.Described biomolecule carrier 20 can be metal, metal nanoparticle, gold size, metal oxide nano-material etc.
The biomolecule carrier 20 of described InP-base HEMT sensor can be designed to grid voltage element and not have grid voltage element two class.By grid voltage element, a grid voltage can be applied to the biomolecule carrier 20 being positioned at grid, thus condition of work can be changed by additional grid voltage selector.
The device layout having grid voltage element as shown in Figure 2, applies bias voltage by grid voltage element 203, changes the condition of work of device.Without grid voltage element device layout as shown in Figure 3.Fig. 2 and Fig. 3 is the vertical view of sensor shown in Fig. 1.
Described bio-sensitive film 30 is for specific recognition tumor markers, and its fixing means can be chemical bond method or physisorphtion.Chemical bond method is by covalent bonding method or Non-covalent binding method fixed biologically sensitive molecule in biomolecule carrier 20, and the chemical substance of use can be chemical substance (mercaptoacetic acid etc.) simultaneously containing sulfydryl and carboxyl, antibody after sulfydryl modification, have the chemical substance (cysteamine etc.) of sulfydryl and other function bases (amino etc.) simultaneously.Physisorption rule is physisorption fixed biologically sensitive molecule in biomolecule carrier 20 directly, and wherein said bio-sensing molecule is the antibody protein etc. corresponding with the tumor markers detected.
Introduce the method for making of the InP-base HEMT tumor marker sensor manufacturing above-described embodiment below, it is as follows that it specifically comprises step:
Step S1: preparation InP-base HEMT.
It comprises stacked substrate 101, cushion 102, delta doping layer 103, first separation layer 104 of a Si, channel layer 105, second separation layer 106, the delta doping layer 107 of the 2nd Si, barrier layer 108, cap layers 109 from bottom to up successively.
This step is prepared by using Veeco GEN-II type MBE system.
Step S2: make source, drain electrode on described InP-base HEMT.
First, carve outside the active area of described InP-base HEMT, etching obtains table top.This step obtains table top by dry etching, the material structure layer that dry etching is removed comprises the delta doping layer 103 of a Si, first separation layer 104, channel layer 105, the delta doping layer 107 of the second separation layer the 106, two Si, barrier layer 108, cap layers 109, until etch into cushion 102, forms the active region mesa at device place.
Then, source, the drain electrode 110 of Ohmic contact is prepared.The Ohmic contact of this step by using the method for electron beam evaporation deposition to prepare nickel germanium gold system, uses acetone soln to carry out metal-stripping, under nitrogen atmosphere protection, and short annealing 20 ~ 50s at 380 DEG C ~ 400 DEG C.
Then, the highly doped cap layers 109 of area of grid between wet etching leakage, drain electrode exposes barrier layer 108.This step is by using Succinic Acid Polyester Polyols corrosive liquid corrosion grid groove, and preparation succinic acid solution, by using between ammoniacal liquor adjust ph to 5.0 ~ 5.6, adds H in the succinic acid solution modulated
2o
2solution.
Finally, the device protecting layer 111 of sex change photoresist is prepared.This step by even glue, front baking, exposure, development, rear baking, beat glue and high temperature and dry the protective seam obtaining photoresist, obtain the InP-base HEMT transducer for tumor marker sensor.
Step S3: the biomolecule carrier making bio-sensitive film at the area of grid of described InP-base HEMT.
The barrier layer 108 that described bio-sensitive film 30 exposes at area of grid for this embodiment of specific recognition tumor markers prepares Au nano particle biomolecule carrier 20.By using spin-coating method in the area of grid spin coating layer of Au nanoparticles solution of InP-base HEMT, and under certain condition, Au nano particle is made to be fixed on area of grid.After this step, draw Au line, for follow-up test by pressure welding in the pressure welding of device electrode place.
Step S4: prepare bio-sensitive film on the bio-sensitive film of area of grid.
A kind of mode of this step be by use cysteamine solution and grid Au nano particle under certain condition (as constant temperature wets in box) react, form S-Au key, secondly by using glutaraldehyde solution, itself and grid cysteamine is made to react bonding, (as constant temperature wets in box) is by using CEA antibody formation bio-sensitive film under certain condition, by using sodium borohydride solution to block the remaining glutaraldehyde of passivation, obtain the CEA biology sensor based on InP-base HEMT.
Another kind of mode is: by using sulfydryl butanediamine aqueous solution, RuCl
3solution and gold size etc. are fixed corresponding antibodies at grid and are obtained bio-sensitive film.Particularly, by device grids Au nano particle is dipped in sulfydryl butanediamine aqueous solution, (as under room temperature) formation self-assembled molecule layer under certain condition; By dripping RuCl to grid
3after solution reaction a period of time, then soak in gold size; Carry out reacting thus fixation of C A125 antibody by drip anti-CA 125 solution room temperature to grid under; Obtain based on InP-base High Electron Mobility Transistor CA125 biology sensor.
The present invention is applicable to alpha-fetoprotein (AFP), oncofetal protein (CEA), pancreas embryo albumen (POA), CA125, CA199, CA153, CA724, CA242, squamous cell carcinoma antigen (SCCA), human chorionic's glandular hormone (HCG), prostate specific antigen (PSA), neuronspecific enolase (NSE), cytokeratin (CK), tissue polypeptide antigen (TPA), ferritin, ras gene protein, the development of the tumor markers biology sensors such as myc gene protein.
As from the foregoing, InP-base HEMT is utilized can to make the biology sensor that can detect various tumor markers.The present invention devises transducer for the InP-base HEMT of tumor marker sensor and corresponding manufacture method, and the tumor markers biology sensor for the high sensitivity of development and the speed of response that is exceedingly fast has important meaning.With the detection tumor marker sensor that InP-base HEMT develops for substrate, can the raising detection limit of high degree, expectation can detect the pg/mL even alpha-fetoprotein of fg/mL in solution, this will provide foundation for the various malignant tumour of preventive assessment, grasp best occasion for the treatment, reduce the mortality ratio of malignant tumour.The present invention can be used for detecting various tumor markers, has important effect for various malignant tumours such as diagnosing, prevent and treat lung cancer, liver cancer, cancer of the stomach, breast cancer clinically.
Above-described specific embodiment; object of the present invention, technical scheme and beneficial effect are further described; be understood that; the foregoing is only specific embodiments of the invention; be not limited to the present invention; within the spirit and principles in the present invention all, any amendment made, equivalent replacement, improvement etc., all should be included within protection scope of the present invention.
Claims (8)
1. an InP-base HEMT tumor marker sensor, for detecting tumor markers, is characterized in that, comprising:
InP-base HEMT transducer, it comprises source electrode, drain and gate;
Biomolecule carrier, is formed at the grid of described transducer;
Bio-sensitive film, be attached to described biomolecule carrier, this bio-sensitive film energy specific recognition tumor markers, be subject to the impact of the electric charge of tumor markers own, described gate surface CHARGE DISTRIBUTION is changed, and then affect two-dimensional electron gas in described transducer raceway groove, the source-drain current of transducer is changed.
2. InP-base HEMT tumor marker sensor as claimed in claim 1, it is characterized in that, described biomolecule carrier is metal, metal nanoparticle, gold size, metal oxide nano-material.
3. InP-base HEMT tumor marker sensor as claimed in claim 1, it is characterized in that, described biomolecule carrier has grid voltage element, and described grid voltage element is for applying voltage.
4. InP-base HEMT tumor marker sensor as claimed in claim 1, it is characterized in that, area of grid area distributions is at 1000 ~ 1500 μm
2between.
5. make a method for InP-base HEMT tumor marker sensor, it is characterized in that, comprise the steps:
Step S1: preparation InP-base HEMT;
Step S2: make source, drain electrode on described InP-base HEMT;
Step S3: the biomolecule carrier making bio-sensitive film at the area of grid of described InP-base HEMT;
Step S4: prepare bio-sensitive film on the bio-sensitive film of area of grid, this bio-sensitive film energy specific recognition tumor markers, be subject to the impact of the electric charge of tumor markers own, described gate surface CHARGE DISTRIBUTION is changed, and then affect two-dimensional electron gas in described transducer raceway groove, the source-drain current of transducer is changed.
6. the method making InP-base HEMT tumor marker sensor as claimed in claim 5, it is characterized in that, described biomolecule carrier is Au nano particle, described step S4 is: react by using cysteamine solution and described biomolecule carrier, form S-Au key, re-use glutaraldehyde solution, make itself and described cysteamine react bonding, by using antibody to form bio-sensitive film, block the remaining glutaraldehyde of passivation by using sodium borohydride solution.
7. the method making InP-base HEMT tumor marker sensor as claimed in claim 6, it is characterized in that, described step S4 is: by using sulfydryl butanediamine aqueous solution, RuCl
3solution and gold size are fixed corresponding antibodies at described grid and are obtained bio-sensitive film.
8. the method making InP-base HEMT tumor marker sensor as claimed in claim 5, it is characterized in that, described biomolecule carrier is Au nano particle, described step S4 is: form self-assembled molecule layer by biomolecule carrier being dipped in sulfydryl butanediamine aqueous solution, then by dripping RuCl to described grid
3after solution reaction a period of time, then soak in gold size, then drip antibody-solutions to grid, carry out under room temperature reacting thus fixing this antibody.
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