Preparation method and application of a kind of competitive aflatoxin immunosensor
An aflatoxin and immunosensor technology, which can be used in instruments, scientific instruments, measuring devices, etc., can solve the problems of complex procedures, long detection periods, and many required reagents, and achieve improved detection sensitivity and good biocompatibility. Effect
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Embodiment 1
[0033] The preparation of the hydroxyapatite-labeled antibody loaded with copper ions according to the present invention, the steps are as follows:
[0034] (1) Mix nano-hydroxyapatite with a particle size of 30nm and copper sulfate at a mass ratio of 1:3, dissolve in 5mL water, shake for 12 hours, and centrifuge;
[0035] (2) Take 5 mg of the obtained nano-hydroxyapatite loaded with copper ions, dissolve it in 5 mL of phosphate buffer solution, add 0.1 ml of aflatoxin antibody solution with a concentration of 1 μg / mL, incubate and shake for 4 hours, and centrifuge;
[0036] (3) Re-add the obtained hydroxyapatite-labeled antibody loaded with copper ions into 5 mL of phosphate buffer solution, and add 1 mg of bovine serum albumin, incubate and shake for 4 hours, and centrifuge;
[0037] (4) Redisperse the precipitate into 5 mL of phosphate buffer solution to obtain the stock solution of the hydroxyapatite-labeled antibody loaded with copper ions.
Embodiment 2
[0039] The preparation of the competitive type aflatoxin immunosensor of the present invention, the steps are as follows
[0040] (1) Add 50μL of an aqueous solution containing 0.05mmol / L silver nitrate, 0.2mmol / L sodium citrate and 0.05mol / L potassium nitrate to the screen-printed electrode, connect to the electrochemical workstation, and perform constant potential deposition at -0.2V, Deposition time is 120s, dries in the air, makes the screen-printed electrode of silver nanoparticle modification;
[0041] (2) Add 10 μL of aflatoxin antibody solution with a concentration of 1 μg / mL dropwise on the screen-printed electrode, incubate for 1 hour, and wash it with phosphate buffer solution;
[0042] (3) Add 10 μL of 1% bovine serum albumin solution dropwise on the screen-printed electrode, incubate for 1 hour, and wash it with phosphate buffer solution;
[0043] (4) Add 10 μL of the mixed solution of aflatoxin and copper ion-loaded hydroxyapatite-labeled antigen dropwise at a v...
Embodiment 3
[0045] A kind of competitive type aflatoxin immunosensor described in any one of embodiment 1~2, is used for the detection of aflatoxin, comprises the following steps:
[0046] (1) Prepare a series of screen-printed electrode immunosensors according to the method described in any one of Examples 1-2 by using different concentrations of aflatoxin;
[0047] (2) Connect the reference electrode, counter electrode and working electrode to the electrochemical workstation, add 50 μL of acetate buffer solution to the electrolytic cell, and then add 5~10 μL of 1% dilute sulfuric acid solution to load copper ions The hydroxyapatite dissolves and releases copper ions;
[0048] (3) Set the potential range to -0.6~0.4V, the deposition potential to -0.8V, and the deposition time to 100~300s, and detect the current response of the modified screen-printed electrode by anodic stripping linear voltammetry;
[0049] (4) Draw a working curve according to the relationship between the obtained cur...
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