CN102253223A - Electrochemical immune sensing method capable of detecting phytohormone beta-3-indoleacetic acid - Google Patents
Electrochemical immune sensing method capable of detecting phytohormone beta-3-indoleacetic acid Download PDFInfo
- Publication number
- CN102253223A CN102253223A CN2011101110518A CN201110111051A CN102253223A CN 102253223 A CN102253223 A CN 102253223A CN 2011101110518 A CN2011101110518 A CN 2011101110518A CN 201110111051 A CN201110111051 A CN 201110111051A CN 102253223 A CN102253223 A CN 102253223A
- Authority
- CN
- China
- Prior art keywords
- iaa
- solution
- immunosensor
- electrochemical
- reagent type
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention relates to an electrochemical immune sensing method capable of detecting phytohormone beta-3-indoleacetic acid. The electrochemical immune sensing method mainly comprises the following steps of: 1) modifying an electrode and preparing an immune sensor; 2) preparing a standard curve; and 3) detecting a sample. The electrochemical immune sensing method has the advantages that: during construction of a sensor interface, an indole-3-acetic acid (IAA) antibody living body is fixed directionally by using yeast cells and the detection lower limit and the sensitivity are improved, so high-sensitivity quantitative determination of IAA is realized. In the method, a linear relation is provided when the concentration of IAA is 1 to 125ng/mL and the detection lower limit is 0.05ng/mL, so the requirement of IAA content determination in a plant sample can be met better; furthermore, the method has the advantages of simpleness, reliable result, lower cost and the like.
Description
Technical field
The present invention relates to biological detecting method, specifically is a kind of electrochemistry immuno-sensing method that detects plant hormone β-3-indolyl acetic acid.
Background technology
IAA is the abbreviation of Indole-3-acetic acid, the chinesization formal name used at school is called β-3-indolyl acetic acid, the biology title is an auxin, is found the earliest a kind of in the tradition five big class plant hormones, also is a kind of plant growth substance with considerable biological significance and using value.β-3-indolyl acetic acid extensively exists in mitogenetic tissues such as the leaf bud of plant and tender leaf.Vegetable cell often utilizes tryptophane to synthesize β-3-indolyl acetic acid.It is prevalent in the plant, and high concentration suppresses the plant growth, and low concentration promotes the plant growth, and is safe and harmless to non-plant, also do not have effect.Just because of IAA important effect is arranged all in biological physiology and agricultural production, and its content is lower, the chemical constitution instability is easily decomposed, and just has crucial meaning so how IAA content is measured accurately.
Summary of the invention
The technical problem to be solved in the present invention is to overcome the deficiencies in the prior art, and IAA content detecting method in a kind of plant sample is provided.The present invention can satisfy the needs that IAA measures preferably, and method is simple, and is highly sensitive fast, provides a kind of new approach for detecting IAA.In order to achieve the above object, the method for IAA content may further comprise the steps in the detection plant sample provided by the invention: at the no reagent type IAA electrochemical immunosensor of gold electrode surfaces preparation; The drawing standard working curve; IAA content in the actual plant sample is detected.The technical solution adopted for the present invention to solve the technical problems can also be further perfect.Described no reagent type IAA electrochemical immunosensor can be prepared by following steps: (1) will handle clean gold electrode and at first immerse in the 0.02 M cysteamine solution 4 ℃ of reaction 6h down, make its surface form fine and close self-assembled monolayer.(2) after washing is dried, at ambient temperature it is immersed in the yeast cells solution of 10 mg/mL and react 4h, take out, washing is dried.(3) the 0.5 mg/mL IAA antibody-solutions (PBS, pH 7.5) that drips 40ml then is in gold electrode surfaces, 37 ℃ of following incubation 1h.Take out washing, dry.(4) last, the nonspecific reaction site on the 10mg/mLBSA solution sealing crystal oscillator surface of employing 30ml.Promptly obtain no reagent type IAA electrochemical immunosensor in this way, modify that preservation is standby down in 4 ℃ in the PBS solution that good immunosensor electrode catheter can immerse the appropriate pH value.Described production standard working curve method is: the working electrode (the no reagent type IAA electrochemical immunosensor that promptly prepares) that will be modified with antibody places antigen (IAA) solution of variable concentrations to hatch 60min (37 ℃), measure its cyclic voltammetry curve under the same conditions, the response current difference is directly proportional with IAA concentration.For obtaining a stable experimental result, the electric current continuous detecting of biosensor surface up to stable state, is obtained stationary value.Change the concentration of IAA standard solution, measure the IAA standard solution of a series of variable concentrations by above-mentioned steps, with the cyclic voltammetric electric current changing value and the IAA concentration drawing standard working curve on immunosensor surface.Described sample determination method is: measure the no reagent type IAA electrochemical immunosensor surface cyclic voltammetric current value for preparing earlier; Get the sample solution of handling well then and get convention amount and drip to the no reagent type IAA electrochemical immunosensor that has prepared, 37 ℃ of environment are reaction 60min down; The cyclic voltammetric electric current continuous detecting on immunosensor surface up to stable state, is obtained stability number.With response current difference and the numerical value of drawing IAA content in the good standard working curve contrast acquisition plant sample solution.The detection principle of IAA electrochemical immunosensor: the present invention is based on and utilizes electrochemical immunosensor to have high sensitivity, characteristics such as easy, quick, reliable, a kind of automated analysis detection system that forms in conjunction with biological recognition system (antigen and antibody specific combination), the fast detecting of IAA content in the realization plant sample.The present invention utilizes self-assembling technique and yeast cells live body immobilization technology, IAA antibody is fixed on the gold electrode surfaces of modifying by by self-assembled monolayer half Guang ammonia film and Electrostatic Absorption live body yeast cells, solved the fixing difficult problem of antibody, on electrode, drip testing sample then, because the immune response of high specific can take place with anti-IAA in IAA, if IAA content height in the sample, the amount of IAA that is attached to gold electrode surfaces is just many, uses cyclic voltammetry then.In fact in response signal and the sample 2,4-D content is directly proportional in the present invention, and promptly IAA is many more in the sample, and the antigen that is connected on the gold electrode is just many more, and immune response electric current difference is just big more, and signal is obvious more.Effect of the present invention is: linear in IAA concentration is the 1ng/mL-125ng/mL scope, calibration curve is y=0.0395x+0.6499, and relative coefficient R is 0.9976, is limited to 0.05ng/mL under detecting.Confirm that through using the method can satisfy the needs of IAA assay in the plant sample preferably.
Claims (4)
1. electrochemistry immuno-sensing method that detects plant hormone β-3-indolyl acetic acid (IAA), it is characterized in that may further comprise the steps: (1) is at the no reagent type IAA electrochemical immunosensor of gold electrode surfaces preparation; (2) production standard working curve; (3) IAA content in the actual plant sample is detected.
2. no reagent type IAA electrochemical immunosensor according to claim 1 is prepared by following steps: will handle clean gold electrode and at first immerse in the 0.02 M cysteamine solution 4 ℃ of reaction 6h down, and make its surface form fine and close self-assembled monolayer; After washing is dried, at ambient temperature it is immersed in the yeast cells solution of 10 mg/mL and react 4h, take out, washing is dried; The 0.5 mg/mL IAA antibody-solutions (PBS, pH 7.5) that drips 40ml then is in gold electrode surfaces, 37 ℃ of following incubation 1h; Take out washing, dry; At last, adopt the nonspecific reaction site on the 10mg/mLBSA solution sealing crystal oscillator surface of 30ml; Promptly obtain no reagent type IAA electrochemical immunosensor in this way, modify that preservation is standby down in 4 ℃ in the PBS solution that good immunosensor electrode catheter can immerse the appropriate pH value.
3. the making of standard working curve according to claim 1, it is characterized in that: the working electrode (the no reagent type IAA electrochemical immunosensor that promptly prepares) that will be modified with antibody places antigen (IAA) solution of variable concentrations to hatch 60min (37 ℃), measure its cyclic voltammetry curve under the same conditions, the response current difference is directly proportional with IAA concentration; For obtaining a stable experimental result, the electric current continuous detecting of biosensor surface up to stable state, is obtained stationary value; Change the concentration of IAA standard solution, measure the IAA standard solution of a series of variable concentrations by above-mentioned steps, with the cyclic voltammetric electric current changing value and the IAA concentration drawing standard working curve on immunosensor surface.
4. the method for IAA content in the detection plant sample according to claim 1 is characterized in that: measure the no reagent type IAA electrochemical immunosensor surface cyclic voltammetric current value for preparing earlier; Get the sample solution of handling well then and get convention amount and drip to the no reagent type IAA electrochemical immunosensor that has prepared, 37 ℃ of environment are reaction 60min down; The cyclic voltammetric electric current continuous detecting on immunosensor surface up to stable state, is obtained stability number; With response current difference and the numerical value of drawing IAA content in the good standard working curve contrast acquisition plant sample solution.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011101110518A CN102253223A (en) | 2011-04-30 | 2011-04-30 | Electrochemical immune sensing method capable of detecting phytohormone beta-3-indoleacetic acid |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2011101110518A CN102253223A (en) | 2011-04-30 | 2011-04-30 | Electrochemical immune sensing method capable of detecting phytohormone beta-3-indoleacetic acid |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102253223A true CN102253223A (en) | 2011-11-23 |
Family
ID=44980600
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2011101110518A Pending CN102253223A (en) | 2011-04-30 | 2011-04-30 | Electrochemical immune sensing method capable of detecting phytohormone beta-3-indoleacetic acid |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102253223A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108152483A (en) * | 2017-11-30 | 2018-06-12 | 苏州峰通光电有限公司 | A kind of immune electrochemical analyser |
| CN111060700A (en) * | 2019-12-31 | 2020-04-24 | 湖南农业大学 | Plant auxin receptor TIR1 modified biosensor and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2646714Y (en) * | 2003-08-05 | 2004-10-06 | 湖南农业大学 | Phytohormone immunity sensor |
| US20070196233A1 (en) * | 2006-02-23 | 2007-08-23 | Fujifilm Corporation | Biosensor and method for immobilizing a physiologically active substance |
| CN101788519A (en) * | 2010-01-26 | 2010-07-28 | 湖南农业大学 | Enzyme sensing determination method for indoleacetic acid content in plant sample |
-
2011
- 2011-04-30 CN CN2011101110518A patent/CN102253223A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN2646714Y (en) * | 2003-08-05 | 2004-10-06 | 湖南农业大学 | Phytohormone immunity sensor |
| US20070196233A1 (en) * | 2006-02-23 | 2007-08-23 | Fujifilm Corporation | Biosensor and method for immobilizing a physiologically active substance |
| CN101788519A (en) * | 2010-01-26 | 2010-07-28 | 湖南农业大学 | Enzyme sensing determination method for indoleacetic acid content in plant sample |
Non-Patent Citations (2)
| Title |
|---|
| 卢海霞: "基于纳米金半网状及复合结构的新型生物传感器研究", 《中国优秀硕士学位论文全文数据库》 * |
| 李春香 等: "基于巯基自组装单层膜的植物生长激素吲哚乙酸电化学免疫传感器的研究", 《化学学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108152483A (en) * | 2017-11-30 | 2018-06-12 | 苏州峰通光电有限公司 | A kind of immune electrochemical analyser |
| CN111060700A (en) * | 2019-12-31 | 2020-04-24 | 湖南农业大学 | Plant auxin receptor TIR1 modified biosensor and preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US10895552B1 (en) | Method for preparing ratiometric electrochemical aptasensor for Vaniline based on nanocomposite modified electrode | |
| CN103940867B (en) | A kind of preparation method of the photoelectricity aptamer sensor for detecting 17 beta estradiols | |
| Ahmad et al. | Highly stable urea sensor based on ZnO nanorods directly grown on Ag/glass electrodes | |
| CN101458223B (en) | Preparation of quantitative rapid detecting sensor of microcapsule algae toxin and applications | |
| CN101655473B (en) | Preparation method of nanogold immunoelectrode | |
| CN103575913B (en) | The urinalysis test card of microdose urine protein/UCr | |
| CN201060198Y (en) | Integrated forming disposable electrochemical sensor for determining blood lead concentration | |
| CN103454426B (en) | Preparation method of nanogold/chitosan-graphene-methylene blue modified immunosensor | |
| Valera et al. | Conductimetric immunosensor for atrazine detection based on antibodies labelled with gold nanoparticles | |
| CN104502593B (en) | The preparation method of the unmarked immunosensor of a kind of galvanochemistry stomach neoplasms tumor markers | |
| CN108120750B (en) | A kind of preparation method and application of zearalanol optical electro-chemistry sensor | |
| CN106770544A (en) | Ni MOF ultrathin nanometers band, synthetic method and its application | |
| CN103558268A (en) | Method for electrochemically detecting concentration of glucose in whole blood through integrated paper based micro-fluidic apparatus | |
| Yang et al. | Ratiometric electrochemical sensor for accurate detection of salicylic acid in leaves of living plants | |
| CN108872343A (en) | A kind of Dopamine Sensor and its preparation and application based on nitrogen-doped graphene | |
| CN104502583A (en) | Carbon nano tube/nanogold composite membrane electrochemical immunosensor and application thereof | |
| CN103163193B (en) | Preparation method of electrochemical sensor for detecting three clenbuterol hydrochloride and application | |
| CN102253223A (en) | Electrochemical immune sensing method capable of detecting phytohormone beta-3-indoleacetic acid | |
| Liang et al. | A Smart Portable Sensor with Ratio and Dual-Mode for In-Situ Detection of Nicotine in Tobacco Leaves | |
| CN106841351B (en) | A kind of molybdenum disulfide nano sheet electrochemical sensor and the preparation method and application thereof | |
| CN106596648B (en) | Preparation method for the bacterium living beings sensor that antibiotic in environment tests and analyzes | |
| Li et al. | A highly sensitive electrochemical impedance immunosensor for indole-3-acetic acid and its determination in sunflowers under salt stress | |
| CN103439273A (en) | Method for carrying out colorimetric detection on glucose by using metal organic coordination compound | |
| CN103196973B (en) | Method for preparing seven-channel aflatoxin immunosensor and application | |
| CN101769892A (en) | Electrochemistry immuno-sensing method for detecting 2,4-dichlorphenoxyacetic acid in agricultural product |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20111123 |
|
| WD01 | Invention patent application deemed withdrawn after publication |