CN105301242A - Preparation method of breast cancer susceptibility gene immune sensor based on flaky strontium ferrite double amplification - Google Patents

Preparation method of breast cancer susceptibility gene immune sensor based on flaky strontium ferrite double amplification Download PDF

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CN105301242A
CN105301242A CN201510794397.0A CN201510794397A CN105301242A CN 105301242 A CN105301242 A CN 105301242A CN 201510794397 A CN201510794397 A CN 201510794397A CN 105301242 A CN105301242 A CN 105301242A
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breast cancer
preparation
susceptibility gene
cancer susceptibility
agsrfe
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任祥
魏琴
吴丹
马洪敏
庞雪辉
闫涛
张勇
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University of Jinan
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    • 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
    • G01N33/57407—Specifically defined cancers
    • G01N33/57415—Specifically defined cancers of breast
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    • 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

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Abstract

The present invention relates to a preparation method of a breast cancer susceptibility gene immune sensor based on flaky strontium ferrite double amplification, and belongs to the technical fields of novel functional materials and biological sensors. According to the present invention, based on the characteristics of large effective specific surface area, excellent catalysis activity and the like of the flaky strontium ferrite, the sensitivity and stability of the immune sensor are significantly improved, and the important significance is provided for early diagnosis of breast cancer.

Description

A kind of preparation method of the breast cancer susceptibility gene immunosensor based on the dual amplification of sheet strontium ferrite
Technical field
The present invention relates to a kind of preparation of the breast cancer susceptibility gene immunosensor based on the dual amplification of sheet strontium ferrite, adopt Ab-AgSrFe 12o 19simultaneously as the label detecting antibody and capture antibody, prepare a kind of electrochemical immunosensor detecting breast cancer susceptibility gene, belong to new function material and bio-sensing detection technique field.
Background technology
Cancer is the general designation of a large class malignant tumour.Tumour is in generation evolution, and tumour cell unconventionality expression or host cell, to the Cucumber produced after tumor response, can be used as tumor markers, for diagnosis and the auxiliary diagnosis of some tumour clinical.Therefore, in clinical research, it is very important for developing a kind of quick, easy, sensitive detection tumor markers method.
Electrochemical immunosensor has that structure is simple, selectivity good, easy and simple to handle, highly sensitive, be easy to miniaturization, can the advantage such as continuous, rapid automatized detection analysis, therefore the present invention has prepared a kind of electrochemical immunosensor based on the dual amplification of sheet strontium ferrite, achieves the detection to breast cancer susceptibility gene.
The clinical testing procedure of current existing tumor markers is a lot, but is all mostly the operating personnel needing main equipment and specialty.Immunosensor is a kind of biology sensor combined with analytical chemistry method by immunological method, by the functionality combination between antigen and antibody, and it the is had advantage such as high sensitivity, high selectivity, analysis be quick and easy and simple to handle.
The present invention adopts N-GSHPCS to be base material, Ab-AgSrFe 12o 19simultaneously as the label detecting antibody and capture antibody, hydrogen peroxide produces electrochemical signals, enhances the sensitivity of sensor, has widened the range of linearity, significantly reduce the detection limit of sensor, achieve the hypersensitive analysis to breast cancer susceptibility gene.The method has that cost is low, highly sensitive, specificity is good, detect the advantages such as quick, and preparation process is comparatively simple, effectively overcomes the deficiency of current tumor-marker object detecting method.
Summary of the invention
1. the preparation based on the breast cancer susceptibility gene immunosensor of the dual amplification of sheet strontium ferrite
(1) by diameter be the glass-carbon electrode Al of 4mm 2o 3burnishing powder is polished, and ultrapure water cleans up, and adds at the electrode surface, dry film forming under room temperature by 6 μ L, 0.1 ~ 3mg/mL nitrogen-doped graphene hydroxypropyl chitosan N-GSHPCS dispersant liquid drop;
(2) the antibody hatching thing Ab-AgSrFe of 6 μ L is dripped 12o 19solution is in electrode surface, and ultrapure water, dries in 4 DEG C of refrigerators;
(3) drip 6 μ L, massfraction be the BSA solution of 0.5 ~ 3.5% in electrode surface, ultrapure water, dries in 4 DEG C of refrigerators;
(4) drip 6 μ L, 0.001 ~ 35ng/mL the breast cancer susceptibility gene antigen standard solution of variable concentrations to electrode surface, ultrapure water, dries in 4 DEG C of refrigerators;
(5) 6 μ L antibody hatching thing Ab-AgSrFe are dripped 12o 19solution is in electrode surface, and ultrapure water, dries in 4 DEG C of refrigerators.
2. the preparation of nitrogen-doped graphene hydroxypropyl chitosan N-GSHPCS dispersion liquid
(1) preparation of nitrogen-doped graphene N-GS
0.5mg graphene oxide is scattered in the DMF of 1 ~ 5mL, after ultrasonic 30min, by the N of graphene oxide, dinethylformamide dispersion liquid heats 0.5 ~ 5h at 153 DEG C, by products obtained therefrom at the centrifugal 10min of 7500 ~ 10000r/min, is separated and obtains N-GS;
(2) preparation of nitrogen-doped graphene hydroxypropyl chitosan N-GSHPCS dispersion liquid
1 ~ 10mgN-GS and 1mL, 0.1 ~ 10mg/mL hydroxypropyl sugar juice are mixed, ultrasonic 1 ~ 10h, obtains N-GSHPCS.
3. antibody hatching thing Ab-AgSrFe 12o 19preparation
(1) AgSrFe 12o 19synthesis
By 0.1gSrFe 12o 19join in 8mL absolute ethyl alcohol, add 0.1 ~ 3mL3-aminopropyl triethoxysilane, 50 ~ 85 DEG C of backflow 2h, centrifuging, vacuum drying, obtained amination SrFe 12o 19; By amidized for 1mg SrFe 12o 19joining 50mL massfraction is in the Ag Nano sol of 1%, and vibration 30h, centrifugal 10min under 7500r/min, be re-dispersed into gained solid in 1mL ultrapure water, obtained AgSrFe 12o 19dispersion liquid;
(2) antibody hatching thing Ab-AgSrFe 12o 19preparation
By 1 ~ 10mgAgSrFe 12o 19join in the breast cancer susceptibility gene antibody A b solution of 1mL, 10 ~ 500 μ g/mL, fully mix, concussion 20h, obtained Ab-AgSrFe 12o 19antibody incubates compound solution.
4. the detection of breast cancer susceptibility gene
(1) use electrochemical workstation to test, contrast electrode is saturated calomel electrode, is platinum electrode to electrode, and prepared immunosensor is working electrode, tests in the PBS buffer solution of 10mL, 10 ~ 200mmoL/L, pH7.00 ~ 7.40;
(2) used time m-current method detects breast cancer susceptibility gene antigen standard solution, and input voltage is-0.6 ~ 0.2V, working time 50 ~ 5000s, sample interval 0.1 ~ 0.6s;
(3) after background current tends towards stability, in the PBS of 10mL, 10 ~ 200mmoL/L, the hydrogen peroxide solution of 10 μ L, 5 ~ 20mol/L is injected every 50s, then record current change, drawing curve.
useful achievement of the present invention
(1) use of N-GSHPCS, increases the specific surface area of electrode surface, can be good at the electron transmission promoting electrode surface simultaneously, and HPCS is can be good at fix N-GS, connecting antibody, improves the stability of sensor.
(2) sheet SrFe 12o 19use, can more multi-link effective breast cancer susceptibility gene antibody, realize the detection to more antigen.
(3) based on Ab-AgSrFe 12o 19for Stationary liquid, construct the electrochemical immunosensor of dual amplification, the detection antibody of this sensor and capture antibody use AgSrFe simultaneously 12o 19hatch, the method is the novel sensor construction method built first.
(4) electrochemical immunosensor prepared of the present invention is for the detection of breast cancer susceptibility gene, response time is short, detectability is low, the range of linearity is wide, simple, quick, highly sensitive and specific detection can be realized, 0.33pg/mL is reached to breast cancer susceptibility gene detection limit, the early warning to breast cancer can be realized.
Embodiment
embodiment 1a kind of preparation of the breast cancer susceptibility gene immunosensor based on the dual amplification of sheet strontium ferrite
(1) by diameter be the glass-carbon electrode Al of 4mm 2o 3burnishing powder is polished, and ultrapure water cleans up, and adds at the electrode surface, dry film forming under room temperature by 6 μ L, 0.1mg/mL nitrogen-doped graphene hydroxypropyl chitosan N-GSHPCS dispersant liquid drops;
(2) the antibody hatching thing Ab-AgSrFe of 6 μ L is dripped 12o 19solution is in electrode surface, and ultrapure water, dries in 4 DEG C of refrigerators;
(3) drip 6 μ L, massfraction be the BSA solution of 0.5% in electrode surface, ultrapure water, dries in 4 DEG C of refrigerators;
(4) drip 6 μ L, 0.001 ~ 35ng/mL the breast cancer susceptibility gene antigen standard solution of variable concentrations to electrode surface, ultrapure water, dries in 4 DEG C of refrigerators;
(5) 6 μ L antibody hatching thing Ab-AgSrFe are dripped 12o 19solution is in electrode surface, and ultrapure water, dries in 4 DEG C of refrigerators.
embodiment 2a kind of preparation of the breast cancer susceptibility gene immunosensor based on the dual amplification of sheet strontium ferrite
(1) by diameter be the glass-carbon electrode Al of 4mm 2o 3burnishing powder is polished, and ultrapure water cleans up, and adds at the electrode surface, dry film forming under room temperature by 6 μ L, 1mg/mL nitrogen-doped graphene hydroxypropyl chitosan N-GSHPCS dispersant liquid drops;
(2) the antibody hatching thing Ab-AgSrFe of 6 μ L is dripped 12o 19solution is in electrode surface, and ultrapure water, dries in 4 DEG C of refrigerators;
(3) drip 6 μ L, massfraction be the BSA solution of 2% in electrode surface, ultrapure water, dries in 4 DEG C of refrigerators;
(4) drip 6 μ L, 0.001 ~ 35ng/mL the breast cancer susceptibility gene antigen standard solution of variable concentrations to electrode surface, ultrapure water, dries in 4 DEG C of refrigerators;
(5) 6 μ L antibody hatching thing Ab-AgSrFe are dripped 12o 19solution is in electrode surface, and ultrapure water, dries in 4 DEG C of refrigerators.
embodiment 3a kind of preparation of the breast cancer susceptibility gene immunosensor based on the dual amplification of sheet strontium ferrite
(1) by diameter be the glass-carbon electrode Al of 4mm 2o 3burnishing powder is polished, and ultrapure water cleans up, and adds at the electrode surface, dry film forming under room temperature by 6 μ L, 0.1 ~ 3mg/mL nitrogen-doped graphene hydroxypropyl chitosan N-GSHPCS dispersant liquid drop;
(2) the antibody hatching thing Ab-AgSrFe of 6 μ L is dripped 12o 19solution is in electrode surface, and ultrapure water, dries in 4 DEG C of refrigerators;
(3) drip 6 μ L, massfraction be the BSA solution of 0.5 ~ 3.5% in electrode surface, ultrapure water, dries in 4 DEG C of refrigerators;
(4) drip 6 μ L, 0.001 ~ 35ng/mL the breast cancer susceptibility gene antigen standard solution of variable concentrations to electrode surface, ultrapure water, dries in 4 DEG C of refrigerators;
(5) 6 μ L antibody hatching thing Ab-AgSrFe are dripped 12o 19solution is in electrode surface, and ultrapure water, dries in 4 DEG C of refrigerators.
embodiment 4the preparation of nitrogen-doped graphene hydroxypropyl chitosan N-GSHPCS dispersion liquid
(1) preparation of nitrogen-doped graphene N-GS
0.5mg graphene oxide is scattered in the DMF of 1mL, after ultrasonic 30min, the DMF dispersion liquid of graphene oxide is heated 0.5h at 153 DEG C, by products obtained therefrom at the centrifugal 10min of 7500r/min, be separated and obtain N-GS;
(2) preparation of nitrogen-doped graphene hydroxypropyl chitosan N-GSHPCS dispersion liquid
1mgN-GS and 1mL, 0.1mg/mL hydroxypropyl sugar juice are mixed, ultrasonic 1h, obtains N-GSHPCS.
embodiment 5the preparation of nitrogen-doped graphene hydroxypropyl chitosan N-GSHPCS dispersion liquid
(1) preparation of nitrogen-doped graphene N-GS
0.5mg graphene oxide is scattered in the DMF of 2mL, after ultrasonic 30min, the DMF dispersion liquid of graphene oxide is heated 2h at 153 DEG C, by products obtained therefrom at the centrifugal 10min of 8500r/min, be separated and obtain N-GS;
(2) preparation of nitrogen-doped graphene hydroxypropyl chitosan N-GSHPCS dispersion liquid
5mgN-GS and 1mL, 5mg/mL hydroxypropyl sugar juice are mixed, ultrasonic 3h, obtains N-GSHPCS.
embodiment 6the preparation of nitrogen-doped graphene hydroxypropyl chitosan N-GSHPCS dispersion liquid
(1) preparation of nitrogen-doped graphene N-GS
0.5mg graphene oxide is scattered in the DMF of 5mL, after ultrasonic 30min, the DMF dispersion liquid of graphene oxide is heated 5h at 153 DEG C, by products obtained therefrom at the centrifugal 10min of 10000r/min, be separated and obtain N-GS;
(2) preparation of nitrogen-doped graphene hydroxypropyl chitosan N-GSHPCS dispersion liquid
10mgN-GS and 1mL, 10mg/mL hydroxypropyl sugar juice are mixed, ultrasonic 10h, obtains N-GSHPCS.
embodiment 7antibody hatching thing Ab-AgSrFe 12o 19preparation
(1) AgSrFe 12o 19synthesis
By 0.1gSrFe 12o 19join in 8mL absolute ethyl alcohol, add 0.1mL3-aminopropyl triethoxysilane, 50 DEG C of backflow 2h, centrifuging, vacuum drying, obtained amination SrFe 12o 19; By amidized for 1mg SrFe 12o 19joining 50mL massfraction is in the Ag Nano sol of 1%, and vibration 30h, centrifugal 10min under 7500r/min, be re-dispersed into gained solid in 1mL ultrapure water, obtained AgSrFe 12o 19dispersion liquid;
(2) antibody hatching thing Ab-AgSrFe 12o 19preparation
By 1mgAgSrFe 12o 19join in the breast cancer susceptibility gene antibody A b solution of 1mL, 10 μ g/mL, fully mix, concussion 20h, obtained Ab-AgSrFe 12o 19antibody incubates compound solution.
embodiment 8antibody hatching thing Ab-AgSrFe 12o 19preparation
(1) AgSrFe 12o 19synthesis
By 0.1gSrFe 12o 19join in 8mL absolute ethyl alcohol, add 2mL3-aminopropyl triethoxysilane, 55 DEG C of backflow 2h, centrifuging, vacuum drying, obtained amination SrFe 12o 19; By amidized for 1mg SrFe 12o 19joining 50mL massfraction is in the Ag Nano sol of 1%, and vibration 30h, centrifugal 10min under 7500r/min, be re-dispersed into gained solid in 1mL ultrapure water, obtained AgSrFe 12o 19dispersion liquid;
(2) antibody hatching thing Ab-AgSrFe 12o 19preparation
By 6mgAgSrFe 12o 19join in the breast cancer susceptibility gene antibody A b solution of 1mL, 50 μ g/mL, fully mix, concussion 20h, obtained Ab-AgSrFe 12o 19antibody incubates compound solution.
embodiment 9antibody hatching thing Ab-AgSrFe 12o 19preparation
(1) AgSrFe 12o 19synthesis
By 0.1gSrFe 12o 19join in 8mL absolute ethyl alcohol, add 3mL3-aminopropyl triethoxysilane, 85 DEG C of backflow 2h, centrifuging, vacuum drying, obtained amination SrFe 12o 19; By amidized for 1mg SrFe 12o 19joining 50mL massfraction is in the Ag Nano sol of 1%, and vibration 30h, centrifugal 10min under 7500r/min, be re-dispersed into gained solid in 1mL ultrapure water, obtained AgSrFe 12o 19dispersion liquid;
(2) antibody hatching thing Ab-AgSrFe 12o 19preparation
By 10mgAgSrFe 12o 19join in the breast cancer susceptibility gene antibody A b solution of 1mL, 500 μ g/mL, fully mix, concussion 20h, obtained Ab-AgSrFe 12o 19antibody incubates compound solution.
embodiment 10the detection of breast cancer susceptibility gene
(1) use electrochemical workstation to test, contrast electrode is saturated calomel electrode, is platinum electrode to electrode, and prepared immunosensor is working electrode, tests in the PBS buffer solution of 10mL, 10mmoL/L, pH7.00;
(2) used time m-current method detects breast cancer susceptibility gene antigen standard solution, and input voltage is-0.6V, working time 50s, sample interval 0.1s;
(3) after background current tends towards stability, in the PBS of 10mL, 10mmoL/L, the hydrogen peroxide solution of 10 μ L, 5mol/L is injected every 50s, then record current change, drawing curve.
embodiment 11the detection of breast cancer susceptibility gene
(1) use electrochemical workstation to test, contrast electrode is saturated calomel electrode, is platinum electrode to electrode, and prepared immunosensor is working electrode, tests in the PBS buffer solution of 10mL, 40mmoL/L, pH7.20;
(2) used time m-current method detects breast cancer susceptibility gene antigen standard solution, and input voltage is-0.2V, working time 500s, sample interval 0.4s;
(3) after background current tends towards stability, in the PBS of 10mL, 50mmoL/L, the hydrogen peroxide solution of 10 μ L, 10mol/L is injected every 50s, then record current change, drawing curve.
embodiment 12the detection of breast cancer susceptibility gene
(1) use electrochemical workstation to test, contrast electrode is saturated calomel electrode, is platinum electrode to electrode, and prepared immunosensor is working electrode, tests in the PBS buffer solution of 10mL, 200mmoL/L, pH7.40;
(2) used time m-current method detects breast cancer susceptibility gene antigen standard solution, and input voltage is 0.2V, working time 5000s, sample interval 0.6s;
(3) after background current tends towards stability, in the PBS of 10mL, 200mmoL/L, the hydrogen peroxide solution of 10 μ L, 20mol/L is injected every 50s, then record current change, drawing curve.

Claims (4)

1., based on a preparation for the breast cancer susceptibility gene immunosensor of the dual amplification of sheet strontium ferrite, it is characterized in that, comprise the following steps:
(1) by diameter be the glass-carbon electrode Al of 4mm 2o 3burnishing powder is polished, and ultrapure water cleans up, and adds at the electrode surface, dry film forming under room temperature by 6 μ L, 0.1 ~ 3mg/mL nitrogen-doped graphene hydroxypropyl chitosan N-GSHPCS dispersant liquid drop;
(2) the antibody hatching thing Ab-AgSrFe of 6 μ L is dripped 12o 19solution is in electrode surface, and ultrapure water, dries in 4 DEG C of refrigerators;
(3) drip 6 μ L, massfraction be the BSA solution of 0.5 ~ 3.5% in electrode surface, ultrapure water, dries in 4 DEG C of refrigerators;
(4) drip 6 μ L, 0.001 ~ 35ng/mL the breast cancer susceptibility gene antigen standard solution of variable concentrations to electrode surface, ultrapure water, dries in 4 DEG C of refrigerators;
(5) 6 μ L antibody hatching thing Ab-AgSrFe are dripped 12o 19solution is in electrode surface, and ultrapure water, dries in 4 DEG C of refrigerators.
2. the preparation method of a kind of breast cancer susceptibility gene immunosensor based on the dual amplification of sheet strontium ferrite as claimed in claim 1, described nitrogen-doped graphene hydroxypropyl chitosan N-GSHPCS dispersion liquid, it is characterized in that, preparation process is as follows:
(1) preparation of nitrogen-doped graphene N-GS
0.5mg graphene oxide is scattered in the DMF of 1 ~ 5mL, after ultrasonic 30min, by the N of graphene oxide, dinethylformamide dispersion liquid heats 0.5 ~ 5h at 153 DEG C, by products obtained therefrom at the centrifugal 10min of 7500 ~ 10000r/min, is separated and obtains N-GS;
(2) preparation of nitrogen-doped graphene hydroxypropyl chitosan N-GSHPCS dispersion liquid
1 ~ 10mgN-GS and 1mL, 0.1 ~ 10mg/mL hydroxypropyl sugar juice are mixed, ultrasonic 1 ~ 10h, obtains N-GSHPCS.
3. the preparation method of a kind of breast cancer susceptibility gene immunosensor based on the dual amplification of sheet strontium ferrite as claimed in claim 1, described antibody hatching thing Ab-AgSrFe 12o 19, it is characterized in that, preparation process is as follows:
(1) AgSrFe 12o 19synthesis
By 0.1gSrFe 12o 19join in 8mL absolute ethyl alcohol, add 0.1 ~ 3mL3-aminopropyl triethoxysilane, 50 ~ 85 DEG C of backflow 2h, centrifuging, vacuum drying, obtained amination SrFe 12o 19; By amidized for 1mg SrFe 12o 19joining 50mL massfraction is in the Ag Nano sol of 1%, and vibration 30h, centrifugal 10min under 7500r/min, be re-dispersed into gained solid in 1mL ultrapure water, obtained AgSrFe 12o 19dispersion liquid;
(2) antibody hatching thing Ab-AgSrFe 12o 19preparation
By 1 ~ 10mgAgSrFe 12o 19join in the breast cancer susceptibility gene antibody A b solution of 1mL, 10 ~ 500 μ g/mL, fully mix, concussion 20h, obtained Ab-AgSrFe 12o 19antibody incubates compound solution.
4. the preparation method of a kind of breast cancer susceptibility gene immunosensor based on the dual amplification of sheet strontium ferrite as claimed in claim 1, for the detection of breast cancer susceptibility gene, detecting step is as follows:
(1) use electrochemical workstation to test, contrast electrode is saturated calomel electrode, is platinum electrode to electrode, and prepared immunosensor is working electrode, tests in the PBS buffer solution of 10mL, 10 ~ 200mmoL/L, pH7.00 ~ 7.40;
(2) used time m-current method detects breast cancer susceptibility gene antigen standard solution, and input voltage is-0.6 ~ 0.2V, working time 50 ~ 5000s, sample interval 0.1 ~ 0.6s;
(3) after background current tends towards stability, in the PBS of 10mL, 10 ~ 200mmoL/L, the hydrogen peroxide solution of 10 μ L, 5 ~ 20mol/L is injected every 50s, then record current change, drawing curve.
CN201510794397.0A 2015-11-18 2015-11-18 Preparation method of breast cancer susceptibility gene immune sensor based on flaky strontium ferrite double amplification Pending CN105301242A (en)

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CN106442675A (en) * 2016-11-01 2017-02-22 济南大学 Preparation and application of carcino-embryonic antigen electrochemical immunosensor based on Au@Ag@Au marker
CN108226252A (en) * 2018-01-19 2018-06-29 山东理工大学 A kind of preparation method and application for the Amperometric Immunosensor for detecting breast cancer

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CN101913855A (en) * 2010-08-24 2010-12-15 中北大学 Preparation method of iron strontium oxide magnetic nanoparticles and magnetic damping rubber thereof
CN103235021A (en) * 2013-04-19 2013-08-07 济南大学 Manufacturing method and application of sensor for simultaneously detecting three breast-cancer tumor markers
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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106442675A (en) * 2016-11-01 2017-02-22 济南大学 Preparation and application of carcino-embryonic antigen electrochemical immunosensor based on Au@Ag@Au marker
CN108226252A (en) * 2018-01-19 2018-06-29 山东理工大学 A kind of preparation method and application for the Amperometric Immunosensor for detecting breast cancer
CN108226252B (en) * 2018-01-19 2020-01-07 山东理工大学 Preparation method and application of current type immunosensor for detecting breast cancer

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