CN108152483A - A kind of immune electrochemical analyser - Google Patents
A kind of immune electrochemical analyser Download PDFInfo
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- CN108152483A CN108152483A CN201711240404.8A CN201711240404A CN108152483A CN 108152483 A CN108152483 A CN 108152483A CN 201711240404 A CN201711240404 A CN 201711240404A CN 108152483 A CN108152483 A CN 108152483A
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- electrochemical analyser
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Classifications
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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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/28—Electrolytic cell components
- G01N27/30—Electrodes, e.g. test electrodes; Half-cells
- G01N27/327—Biochemical electrodes, e.g. electrical or mechanical details for in vitro measurements
- G01N27/3275—Sensing specific biomolecules, e.g. nucleic acid strands, based on an electrode surface reaction
Abstract
The invention belongs to biomedicine technical fields, and in particular to a kind of immune electrochemical analyser, preparation method and the usage.The saccharomycete with antigen-binding activity is connected on the working electrode of the electrochemical analyser, the electrochemical analyser can quickly, accurate, efficient detection blood, urine or the HER2 albumen on circulating tumor cell surface, this method high specificity, accuracy are high, and detectable concentration range is up to 10‑9mol/L.The present invention also provides a kind of method for detecting tumor marker HER2, there is high precision, the high and low cost of repeatability, easy to operate, suitable for all kinds of tumour coherent detections.
Description
Technical field
The present invention relates to field of immunodetection, more particularly, to a kind of immune electrochemical analyser and preparation method thereof and use
On the way.
Background technology
ErbB-2 (human epidermal growth factor receptor-2, HER2,
Also known as ErbB2) gene, on chromosome 17q12-21.32, encode the transmembrane receptor sample that relative molecular mass is 185000
Albumen has tyrosine kinase activity, is one of member of human epidermal growth factor acceptor family.It is with important signal
Transduction is important Prognosis in Breast Cancer and judges the factor, and HER2 is positive（It is overexpressed or expands）Breast cancer, clinical characters
There are Special Manifestations with biological characteristics, treatment mode also makes a big difference with other kinds of breast cancer, and accordingly, HER2's has
Effect detection is extremely important to efficient diagnosis and treatment breast cancer.
The detection of HER2 at present predominantly detects method and includes tissue HER2 detections and Serum HER2 detection, wherein, tissue
HER2 is detected and is mainly applied fluorescence in situ hybridization technique, chromogenic in situ hybridization and immunohistochemistry staining method, and serum
HER2 detects main applied chemistry luminescence method detection albumen extracellular fragment.
Immunohistochemistry staining method（immunohistochemistry, IHC）It is the specificity knot using antigen-antibody
Reaction is closed to detect a special kind of skill with certain chemical substances in position tissue and cell, most of Pathology Labs use
The method is detected HER2, and IHC methods have very fast, easier advantage, can realize automation mechanized operation, immunohistochemistry
Slice preservation is more long, but its testing result is easily influenced by prison step and generates bias, belongs to semi-quantitative detection method, detection knot
The reading of fruit is more subjective.
Fluorescence in situ hybridization technique（Fluorescence in situ hybridization, FISH）It is known to
The special DNA sequence dna of population in microorganism different classifications rank, using the specific oligos segment by the use of fluorescent marker as
Probe detects the amplification of HER2 genes.The method has higher precision relative to IHC methods, can automation mechanized operation and stability
It is higher, but its is expensive, and signal strength is weak at any time.
Pigment in situ hybridization (Chromogenicin Situ Hybridization, CISH) refers to HER2 gene probes
With digoxin or biotin labeling, using the reaction of peroxidase or alkaline phosphatase, tissue is detected under an optical microscope
The expression of mRNA has the advantages that dying-stable, but it belongs to more novel technology, experimenter is operated horizontal
More demanding, popularization degree is poor.
HER2 is important tumor biomarker.In blood HER2 segments detection can as breast cancer detection and
The important indicator of prognosis.HER2 can also be used as the tumor biomarker of carcinoma of urinary bladder in urine sample.Existing detection means is general
It is that tumor tissues is taken to carry out immunostaining or blood is taken to carry out ELISA inspections.The requirement of former approach technology is high, samples misdiagnosis rate
Height, and have certain wound to patient.Latter method has that time-consuming and can not monitor in real time.For lacking for the prior art
It falls into, the present invention provides a kind of method of quick, accurate detection HER2, is required to patient's hurtless measure, to experiment operator low, accurate
Exactness is high, misdiagnosis rate is low, suitable for quick detection demand.
Invention content
In order to solve the above-mentioned technical problem, realize that HER2 quickly, is accurately detected, the present invention provides one kind and electrification credit is immunized
Analyzer is connected with saccharomycete on the working electrode of the electrochemical analyser.
Further, the saccharomycete is the S. cervisiae of genetic engineering transformation.
Further, the saccharomycete includes surface display system.
Further, the surface display system includes surface polypeptide sequence, and the surface polypeptide sequence is with antigen
With reference to the amino acid sequence of activity, there is the surface polypeptide sequence electrode to combine activity, it is preferable that the surface polypeptide sequence
Activity is combined with metal electrode.
Further, the amino acid sequence is selected from SEQ ID NO. 6 or SEQ ID NO. 7, and the amino acid can wrap
Containing one or more polypeptides, the polypeptide includes antigen recognizing polypeptide and/or antibody, the antigen recognizing polypeptide and/or antibody
N-terminal, C positioned at the polypeptide section are intermediate.
Further, the preparation method of the yeast surface display system includes：
Polynucleotide sequence or coding corresponding to above-mentioned surface polypeptide sequence will be encoded corresponding to SEQ ID NO. 6 or SEQ
The polynucleotide sequence of ID NO. 7 is connected in cloning vector, obtains recombinant cloning vector pHE1；
PHE1 plasmids directly extract base with LiCl methods or electric robin transformed saccharomyces cerevisiae, the picking His+ transformants on MD tablets
Because group DNA is as template, design primer carries out PCR amplification, and obtained recombinant bacterium E3, positive colony bacterial strain E3 are stored in -80C ice
Case；
After specific density being grown into during the positive colony bacterial strain E3 expression in SD-His fluid nutrient mediums after bacterial strain is recovered
24 hours inducible proteins in the inducing culture containing Galatose are changed to express and be presented in cell surface.
Further, the preparation method of the working electrode includes：Yeast after expression is placed in containing the molten of PBS and BSA
In liquid, working electrode is immersed in above-mentioned solution 30 minutes to 24 hours, it is therefore preferable to 18 hours, the antibody on saccharomycete surface
It is adsorbed by GBP and His-Tag in working electrode surface, saccharomycete immunoelectrode is made and is impregnated 1 hour in 1% BSA solution
To close heterogenetic antigen binding site.
Preferably, the working electrode is metal electrode.
Further, the electrochemical analyser is used to detect albumen.
Further, the albumen includes but not limited to tumor marker.
The present invention provides a kind of method of quick, accurate detection HER2, to patient's hurtless measure, to experiment operator requirement
It is low, accuracy is high, misdiagnosis rate is low, suitable for quick detection demand.
Description of the drawings
Fig. 1 is the surface polypeptide sequential structure schematic diagram of electrochemical analyser provided by the invention；
Fig. 2 is the expression vector pHE1 structure diagrams of electrochemical analyser provided by the invention；
Fig. 3 is the yeast surface display system structure diagram of electrochemical analyser provided by the invention；
Fig. 4 is the structure diagram of electrochemical analyser provided by the invention.
Specific embodiment
Embodiment 1 obtains nucleotide sequence
The surface polypeptide sequence of yeast surface display system includes the Aga2, HER2, GBP of sequence link, His-Tag sequences, institute
Coding Aga2, HER2, GBP are stated, the amino acid sequence of the protein of His-Tag passes through GS amino acid sequences（Such as SEQ ID NO.
Shown in 5）Connection, Aga2 are located at 5 ' ends of surface polypeptide sequence（As shown in Figure 1a, surface polypeptide sequence 1, sequence such as SEQ ID
NO. shown in 6）Or positioned at 3 ' ends (shown in Fig. 1 b, surface polypeptide sequence 2, sequence is as shown in SEQ ID NO. 7), Aga2
（97 amino acid）The amino acid sequence of protein is as shown in SEQ ID NO.1 in sequence table；HER2 sections of amino acid sequence choosing
From can be with any antibody or antibody fragment of HER2 antigen bindings（128 amino acid）, sequence is as shown in SEQ ID NO. 2；
The amino acid sequence of GBP protein（12 amino acid）, as shown in SEQ ID NO. 3 in sequence table；His-Tag protein
Amino acid sequence（6-10 amino acid）, as shown in SEQ ID NO. 4 in sequence table, the above-mentioned corresponding nucleosides of amino acid sequence
Acid sequence is synthesized by ThermoFisher companies.
The structure of embodiment 2, yeast surface display system
The nucleotide sequence of the surface polypeptide sequence of synthesis is connected into cloning vector pRS313（ATCC buy) digestion position
Between point HindIII and BamHI, recombinant cloning vector pRS313 is obtained, carrier structure schematic diagram is as shown in Figure 2.Digestion and
Nucleotide sequence is correctly inserted into described in sequence verification recombinant cloning vector pRS313.
The present embodiment used yeast bacterial strain for saccharomyces cerevisiae (Saccharomyces cerevisiae) (MATa AGA1::
GAL1-AGA1::URA3 ura3-52 trp1 leu2-delta200 his3-delta200 pep4::HIS3 prbd1.6R
can1 GAL)（Opportunistic pathogen strain is purchased from ATCC companies）, pRS313 plasmids are carried out using restriction enzyme HindIII and BamHI
After linearisation, with LiCl method transformed saccharomyces cerevisiae INVSc1, the picking His+ transformants on MD tablets, extraction genomic DNA is made
For template, design primer carries out PCR amplification, as a result proves that the nucleotide sequence of surface peptide sequence has been integrated into saccharomyces cerevisiae
In genome, obtained recombinant bacterium is named as E3.
After above-mentioned carrier is transferred to saccharomycete, positive colony bacterial strain E3 is stored in -80C refrigerators.During expression bacterial strain recovery after
After certain density is grown into SD-His fluid nutrient mediums（OD600=1）It changes to 24 in the inducing culture containing Galatose
Hour inducible protein expresses and is presented in cell surface.
Embodiment 3, the preparation for detecting the sensor electrode of HER2 albumen
Yeast after expression is placed in the solution containing PBS and BSA, and working electrode, that is, gold electrode is immersed in above-mentioned solution 30 minutes
To 24 hours, it is therefore preferable to which 18 hours, the antibody on saccharomycete surface was adsorbed by GBP and His-Tag in working electrode surface（Such as
Shown in Fig. 3）, saccharomycete immunoelectrode is made, 1 hour is impregnated in 1% BSA solution to close heterogenetic antigen binding site.
The immunoelectrode production method need not purify antibody, simple and easy to do.The antibody for being illustrated in live body saccharomycete surface is steady
It spends calmly, it is not degradable, and can effectively show antigen antibody binding sites.Saccharomycete or the antibody of expression can also pass through it
Its albumen or microorganism fixation are fixed on electrode, and electrode is selected from the various forms of work electricity such as gold electrode or glass-carbon electrode
Pole.
The detection of HER2 albumen in embodiment 4, biological sample
Electrochemical analyser model CHI760E used in electrochemical sensor system（Purchased from Shanghai Chen Hua company）.Exempted from above-mentioned
The electrochemical sensor formed based on epidemic disease electrode can use cyclic voltammetry, and current method, impedance method, capacitance method is to anti-in sample
Original is quantitative determined.The high affinity of the immunosensor combination antigen-antibody and electrochemical methods it is highly sensitive
Degree, the corresponding time is fast, not by sample turbidity, the advantages that Color influences, and Her2 that can be in the detection sample of specificity, current method
Monitoring lower-cut is up to 10^-9 mol/L.The electrochemical analyser is as shown in Figure 4.
Embodiment 5, biological analyser are used and are resetted
It can be regenerated with reference to the saccharomycete immunoelectrode of Her2 antigens by 10mM pH2.0 Glycine solution, 4 DEG C are stored in and contain
In the PBS solution of 5-10% glycerine.The saccharomycete immunoelectrode modified can be preserved two weeks or more and be become without apparent activity
Change.It should know, the present invention covers the saccharomycete immunoelectrode of antibody known to all sequences, is related to the exhibition of saccharomycete surface
Show that the immunoelectrode of antibody is within the scope of the present invention.
The above is only the preferred embodiment of the present invention, protection scope of the present invention is not limited merely to above-described embodiment,
It is within the scope of the present invention with various process programs of the present inventive concept without substantial differences.
Sequence table
<110>Suzhou Feng Tong Photoelectric Co., Ltd.s
<120>A kind of immune electrochemical analyser
<160> 7
<170> SIPOSequenceListing 1.0
<210> 1
<211> 87
<212> PRT
<213>Artificial sequence (Artificial Sequence)
<220>
<221> PEPTIDE
<223>The amino acid sequence of Aga2 protein
<400> 1
Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val
1 5 10 15
Leu Ala Gln Glu Leu Thr Thr Ile Cys Glu Gln Ile Pro Ser Pro Thr
20 25 30
Leu Glu Ser Thr Pro Tyr Ser Leu Ser Thr Thr Thr Ile Leu Ala Asn
35 40 45
Gly Lys Ala Met Gln Gly Val Phe Glu Tyr Tyr Lys Ser Val Thr Phe
50 55 60
Val Ser Asn Cys Gly Ser His Pro Ser Thr Thr Ser Lys Gly Ser Pro
65 70 75 80
Ile Asn Thr Gln Tyr Val Phe
85
<210> 2
<211> 128
<212> PRT
<213>Artificial sequence (Artificial Sequence)
<220>
<221> PEPTIDE
<223>The amino acid sequence of HER2 antibody
<400> 2
Ala Ser Pro Val Thr Ser Ile Ile Ala Ala Val Val Gly Ile Leu Leu
1 5 10 15
Val Val Val Val Gly Leu Val Leu Gly Ile Leu Ile Lys Arg Arg Arg
20 25 30
Gln Lys Ile Arg Lys Tyr Thr Met Arg Arg Leu Leu Gln Glu Thr Glu
35 40 45
Leu Val Glu Pro Leu Thr Pro Ser Gly Ala Met Pro Asn Gln Ala Gln
50 55 60
Met Arg Ile Leu Lys Glu Thr Glu Leu Arg Lys Val Lys Val Leu Gly
65 70 75 80
Ser Gly Ala Phe Gly Thr Val Tyr Lys Gly Ile Trp Ile Pro Asp Gly
85 90 95
Glu Asn Val Lys Ile Pro Val Ala Ile Lys Val Leu Arg Glu Asn Thr
100 105 110
Ser Pro Lys Ala Asn Lys Glu Ile Leu Asp Glu Ala Tyr Val Met Ala
115 120 125
<210> 3
<211> 13
<212> PRT
<213>Artificial sequence (Artificial Sequence)
<220>
<221> PEPTIDE
<223>The amino acid sequence of GBP polypeptides
<400> 3
Leu Lys Ala His Leu Pro Pro Ser Ser Arg Leu Pro Ser
1 5 10
<210> 4
<211> 6
<212> PRT
<213>Artificial sequence (Artificial Sequence)
<220>
<221> PEPTIDE
<223>The amino acid sequence of His-Tag protein
<400> 4
His His His His His His
1 5
<210> 5
<211> 15
<212> PRT
<213>Artificial sequence (Artificial Sequence)
<220>
<221> PEPTIDE
<223>The amino acid sequence of GSlinker polypeptides
<400> 5
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
1 5 10 15
<210> 6
<211> 270
<212> PRT
<213>Artificial sequence (Artificial Sequence)
<220>
<221> PEPTIDE
<223>Surface polypeptide sequence 1
<400> 6
Met Gln Leu Leu Arg Cys Phe Ser Ile Phe Ser Val Ile Ala Ser Val
1 5 10 15
Leu Ala Gln Glu Leu Thr Thr Ile Cys Glu Gln Ile Pro Ser Pro Thr
20 25 30
Leu Glu Ser Thr Pro Tyr Ser Leu Ser Thr Thr Thr Ile Leu Ala Asn
35 40 45
Gly Lys Ala Met Gln Gly Val Phe Glu Tyr Tyr Lys Ser Val Thr Phe
50 55 60
Val Ser Asn Cys Gly Ser His Pro Ser Thr Thr Ser Lys Gly Ser Pro
65 70 75 80
Ile Asn Thr Gln Tyr Val Phe Gly Gly Gly Gly Ser Gly Gly Gly Gly
85 90 95
Ser Gly Gly Gly Gly Ser Ala Ser Pro Val Thr Ser Ile Ile Ala Ala
100 105 110
Val Val Gly Ile Leu Leu Val Val Val Val Gly Leu Val Leu Gly Ile
115 120 125
Leu Ile Lys Arg Arg Arg Gln Lys Ile Arg Lys Tyr Thr Met Arg Arg
130 135 140
Leu Leu Gln Glu Thr Glu Leu Val Glu Pro Leu Thr Pro Ser Gly Ala
145 150 155 160
Met Pro Asn Gln Ala Gln Met Arg Ile Leu Lys Glu Thr Glu Leu Arg
165 170 175
Lys Val Lys Val Leu Gly Ser Gly Ala Phe Gly Thr Val Tyr Lys Gly
180 185 190
Ile Trp Ile Pro Asp Gly Glu Asn Val Lys Ile Pro Val Ala Ile Lys
195 200 205
Val Leu Arg Glu Asn Thr Ser Pro Lys Ala Asn Lys Glu Ile Leu Asp
210 215 220
Glu Ala Tyr Val Met Ala Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser
225 230 235 240
Gly Gly Gly Gly Ser His His His His His His Leu Lys Ala His Leu
245 250 255
Pro Pro Ser Ser Arg Leu Pro Ser His His His His His His
260 265 270
<210> 7
<211> 255
<212> PRT
<213>Artificial sequence (Artificial Sequence)
<220>
<221> PEPTIDE
<223>Surface polypeptide sequence 2
<400> 7
Ala Ser Pro Val Thr Ser Ile Ile Ala Ala Val Val Gly Ile Leu Leu
1 5 10 15
Val Val Val Val Gly Leu Val Leu Gly Ile Leu Ile Lys Arg Arg Arg
20 25 30
Gln Lys Ile Arg Lys Tyr Thr Met Arg Arg Leu Leu Gln Glu Thr Glu
35 40 45
Leu Val Glu Pro Leu Thr Pro Ser Gly Ala Met Pro Asn Gln Ala Gln
50 55 60
Met Arg Ile Leu Lys Glu Thr Glu Leu Arg Lys Val Lys Val Leu Gly
65 70 75 80
Ser Gly Ala Phe Gly Thr Val Tyr Lys Gly Ile Trp Ile Pro Asp Gly
85 90 95
Glu Asn Val Lys Ile Pro Val Ala Ile Lys Val Leu Arg Glu Asn Thr
100 105 110
Ser Pro Lys Ala Asn Lys Glu Ile Leu Asp Glu Ala Tyr Val Met Ala
115 120 125
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser His
130 135 140
His His His His His Leu Lys Ala His Leu Pro Pro Ser Ser Arg Leu
145 150 155 160
Pro Ser His His His His His His Met Gln Leu Leu Arg Cys Phe Ser
165 170 175
Ile Phe Ser Val Ile Ala Ser Val Leu Ala Gln Glu Leu Thr Thr Ile
180 185 190
Cys Glu Gln Ile Pro Ser Pro Thr Leu Glu Ser Thr Pro Tyr Ser Leu
195 200 205
Ser Thr Thr Thr Ile Leu Ala Asn Gly Lys Ala Met Gln Gly Val Phe
210 215 220
Glu Tyr Tyr Lys Ser Val Thr Phe Val Ser Asn Cys Gly Ser His Pro
225 230 235 240
Ser Thr Thr Ser Lys Gly Ser Pro Ile Asn Thr Gln Tyr Val Phe
245 250 255
Claims (10)
1. a kind of immune electrochemical analyser, which is characterized in that be connected with yeast on the working electrode of the electrochemical analyser
Bacterium.
2. electrochemical analyser according to claim 1, which is characterized in that the saccharomycete is the wine of genetic engineering transformation
Brewer yeast bacterium.
3. electrochemical analyser according to claim 2, which is characterized in that the saccharomycete includes surface display system.
4. electrochemical analyser according to claim 3, which is characterized in that the surface display system includes surface polypeptide
Sequence, the surface polypeptide sequence are the amino acid sequence with antigen-binding activity, and the surface polypeptide sequence has electrode
With reference to activity, it is preferable that there is the surface polypeptide sequence metal electrode to combine activity.
5. electrochemical analyser according to claim 4, which is characterized in that the amino acid sequence is selected from SEQ ID NO.
6 or SEQ ID NO. 7, the amino acid may include one or more polypeptides, the polypeptide include antigen recognizing polypeptide and/or
Antibody, the antigen recognizing polypeptide and/or antibody are located at the N-terminal of the polypeptide, C sections or centre.
6. according to any electrochemical analysers of claim 3-5, which is characterized in that the yeast surface display system
Preparation method includes：
By coding corresponding to the polynucleotide sequence of surface polypeptide sequence described in claim 4 or coding corresponding to SEQ ID NO.
The polynucleotide sequence of 6 or SEQ ID NO. 7 is connected in cloning vector, obtains recombinant cloning vector pHE1；
PHE1 plasmids directly extract base with LiCl methods or electric robin transformed saccharomyces cerevisiae, the picking His+ transformants on MD tablets
Because group DNA is as template, design primer carries out PCR amplification, and obtained recombinant bacterium E3, positive colony bacterial strain E3 are stored in -80C ice
Case；
After specific density being grown into during the positive colony bacterial strain E3 expression in SD-His fluid nutrient mediums after bacterial strain is recovered
24 hours inducible proteins in the inducing culture containing Galatose are changed to express and be presented in cell surface.
7. electrochemical analyser according to claim 6, which is characterized in that the preparation method of the working electrode includes：
Yeast after expression is placed in the solution containing PBS and BSA, working electrode is immersed in 30 minutes to 24 small in above-mentioned solution
When, it is therefore preferable to 18 hours, the antibody on saccharomycete surface was adsorbed by GBP and His-Tag in working electrode surface, and yeast is made
Bacterial immunity electrode impregnates 1 hour to close heterogenetic antigen binding site in 1% BSA solution.
8. electrochemical analyser according to claim 6, which is characterized in that the working electrode is metal electrode.
9. according to any electrochemical analysers of claim 1-8, which is characterized in that the electrochemical analyser is used to examine
Survey albumen.
10. electrochemical analyser according to claim 9, which is characterized in that the albumen includes tumor marker.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711240404.8A CN108152483A (en) | 2017-11-30 | 2017-11-30 | A kind of immune electrochemical analyser |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711240404.8A CN108152483A (en) | 2017-11-30 | 2017-11-30 | A kind of immune electrochemical analyser |
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| CN108152483A true CN108152483A (en) | 2018-06-12 |
Family
ID=62469152
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| CN201711240404.8A Pending CN108152483A (en) | 2017-11-30 | 2017-11-30 | A kind of immune electrochemical analyser |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110907416A (en) * | 2019-11-05 | 2020-03-24 | 中山大学 | Circulating tumor cell detection device based on hollow nano needle tube electroporation system and detection method thereof |
| CN114113255A (en) * | 2021-11-19 | 2022-03-01 | 江南大学 | Electrochemical sensor and preparation and application thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040053425A1 (en) * | 2002-04-19 | 2004-03-18 | Baylor College Of Medicine | Quantitative measurement of proteins using genetically-engineeredglucose oxidase fusion molecules |
| CN101672814A (en) * | 2009-09-27 | 2010-03-17 | 上海大学 | Electrochemical receptor biosensor and application thereof |
| CN102253223A (en) * | 2011-04-30 | 2011-11-23 | 湖南农业大学 | Electrochemical immune sensing method capable of detecting phytohormone beta-3-indoleacetic acid |
| WO2012097081A3 (en) * | 2011-01-11 | 2012-11-15 | The Governing Council Of The University Of Toronto | Protein detection method |
| CN104870471A (en) * | 2013-01-03 | 2015-08-26 | 默克专利股份公司 | Method for producing secretable antibodies by expression in saccharomyces cerevisiae |
| CN107064258A (en) * | 2017-02-20 | 2017-08-18 | 中南大学 | The method that electric signal and its electrochemical aptamer sensor measure HER2 of self assembly amplified signal are produced based on DNA |
-
2017
- 2017-11-30 CN CN201711240404.8A patent/CN108152483A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040053425A1 (en) * | 2002-04-19 | 2004-03-18 | Baylor College Of Medicine | Quantitative measurement of proteins using genetically-engineeredglucose oxidase fusion molecules |
| CN101672814A (en) * | 2009-09-27 | 2010-03-17 | 上海大学 | Electrochemical receptor biosensor and application thereof |
| WO2012097081A3 (en) * | 2011-01-11 | 2012-11-15 | The Governing Council Of The University Of Toronto | Protein detection method |
| CN102253223A (en) * | 2011-04-30 | 2011-11-23 | 湖南农业大学 | Electrochemical immune sensing method capable of detecting phytohormone beta-3-indoleacetic acid |
| CN104870471A (en) * | 2013-01-03 | 2015-08-26 | 默克专利股份公司 | Method for producing secretable antibodies by expression in saccharomyces cerevisiae |
| CN107064258A (en) * | 2017-02-20 | 2017-08-18 | 中南大学 | The method that electric signal and its electrochemical aptamer sensor measure HER2 of self assembly amplified signal are produced based on DNA |
Non-Patent Citations (5)
| Title |
|---|
| LANG Q ET AL.,: "Co-immobilization of glucoamylase and glucose oxidase for electrochemical sequential enzyme electrode for starch biosensor and biofuel cell", 《BIOSENSORS ANDBIOELECTRONICS》 * |
| WANG H ET AL.,: "Electrochemical Glucose Biosensor Based on Glucose Oxidase Displayed on Yeast Surface", 《YEAST SURFACE DISPLAY》 * |
| WANG H ET AL.,: "Yeast surface displaying glucose oxidase as whole-cell biocatalyst: construction, characterization, and its electrochemical glucose sensing application", 《ANALYTICAL CHEMISTRY》 * |
| 宋燕 等: "利用a凝集素在酿酒酵母表面展示CALB 及工业化培养基发酵条件优化", 《安徽农业科学》 * |
| 沈桂芳，丁仁瑞主编: "《现代生物技术与21世纪农业》", 31 December 2000, 杭州：浙江科学技术出版社 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110907416A (en) * | 2019-11-05 | 2020-03-24 | 中山大学 | Circulating tumor cell detection device based on hollow nano needle tube electroporation system and detection method thereof |
| CN114113255A (en) * | 2021-11-19 | 2022-03-01 | 江南大学 | Electrochemical sensor and preparation and application thereof |
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Application publication date: 20180612 |