Electrochemical immunosensor for detecting CP4-EPSPS protein in crops

A CP4-EPSPS and immunosensor technology, applied in the field of herbicide-resistant protein detection, can solve the problems of quantitative analysis of mixed protein samples, single target protein sample as the measurement object, etc., and achieve satisfactory detection sensitivity, good signal response, good The effect of conductivity

Active Publication Date: 2020-12-18
SHANGHAI ACAD OF AGRI SCI
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Problems solved by technology

[0006] Chinese invention patent CN107356646 discloses an electrochemical immunosensor for detecting CP4-EPSPS protein, which uses a composite material of mesoporous carbon and colloidal gold to modify the electrode, thionine The mixed solution with the antibody is coupled to the composite material, and it is used for the detection of CP4-EPSPS protein after blocking with BSA. This scheme only takes a single target protein sample as the measurement object, and cannot quantitatively analyze the mixed protein samples in crops.

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  • Electrochemical immunosensor for detecting CP4-EPSPS protein in crops

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Embodiment

[0049] Preparation method and application of electrochemical immunosensor for detecting CP4-EPSPS protein

[0050] 1) Preparation of colloidal gold

[0051] Prepare colloidal gold by trisodium citrate reduction method: take 1mL 1% chloroauric acid, add it to 99mL double distilled water, heat and boil, accurately absorb 1% trisodium citrate 1mL, add it quickly under the condition of magnetic stirring, when After the color of the liquid turns into wine red, continue heating for 15 minutes to obtain colloidal gold with a particle size of 40 nm; seal it aseptically and store it in the dark at 4°C.

[0052] The prepared colloidal gold is transparent wine red, without precipitation and impurities. After 2 weeks at 4°C, it is observed with the naked eye. There is no precipitation and coagulation, and there is no adhesion on the container wall. The maximum absorption peak of colloidal gold measured by NanoDrop is at 525nm, the particle size is 40nm.

[0053] 2) Preparation of gold-l...

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Abstract

An electrochemical immunosensor for detecting CP4-EPSPS protein in crops comprises a glassy carbon electrode and a recognition element, the surface of the glassy carbon electrode is modified by a composite material, and the composite material is formed by ultrasonic treatment of graphene and amino-rich PAMAM; a gold-labeled antibody is used as a recognition element, a colloidal gold-labeled monoclonal antibody CP4mAb is connected to a composite material on the surface of a glassy carbon electrode in a covalent binding mode, and the electrochemical immunosensor is constructed after 5% BSA sealing. According to the invention, the introduction of gold nanoparticles improves the immobilization amount of an electroactive substance and amplifies a detection signal; The sensor is high in detection specificity and sensitivity for herbicide-resistant protein CP4-EPSPS, simple to operate, quick in reaction, high in sensitivity and good in stability, realizes quantitative detection for trace CP4-EPSPS antigen, and provides a new thought for high-sensitivity quantitative analysis of transgenic crops.

Description

technical field [0001] The invention belongs to the detection field of herbicide-resistant protein, and in particular relates to an electrochemical immunosensor for detecting CP4-EPSPS protein in crops. Background technique [0002] As of 2018, the planted area of ​​GM crops reached 197.1 million hectares, a 100-fold increase compared to 1996. Among the strains of genetically modified crops, herbicide-tolerant crops are an important strain of genetically modified crops, which are constructed by inserting the exogenous CP4-epsps gene into the genome of the crops, so that the crops have certain resistance to glyphosate pesticides. tolerance. [0003] While the rapid development of transgenic technology has brought convenience to agricultural production, it also poses certain threats to the environment. For example, transgenic proteins can be released into the soil through roots, making non-target crops resistant to glyphosate pesticides. At the same time, illegal planting an...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/327G01N27/30G01N33/531G01N33/543G01N33/573
CPCG01N27/3278G01N27/3277G01N27/308G01N33/531G01N33/5438G01N33/573G01N2333/91182
Inventor 曾海娟唐雪明王金斌杨倩雯
Owner SHANGHAI ACAD OF AGRI SCI
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