Photoelectrochemical immunosensor for determining alpha fetoprotein content and preparation method and application thereof
A technology of immunosensor and alpha-fetoprotein, which is applied in the field of photoelectrochemical immunosensor and its preparation for determining the content of alpha-fetoprotein, can solve the problems of poor coupling ability of biomolecules and low electronic conductivity of fullerene, and achieve biophase Good capacitive, good aqueous solution dispersion, high sensitivity effect
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Embodiment 1
[0042] Example 1: AC 60 - Photocurrent-potential map of the enhanced signal of Gr-GO nanocomposite
[0043] 1. Instrument: Shanghai Chenhua Electrochemical Workstation (chi 600e software); 150 watt xenon lamp
[0044] 2. Material: Indium tin oxide transparent conductive glass electrode (ITO); AC 60 ;AC 60 -Gr;Gr-GO;AC 60 -Gr-GO nanocomposites with AC 60 is alkylated fullerene, Gr is graphite flake, GO is graphene oxide, AC 60 -Gr-GO nanocomposite is an alkylated fullerene-graphite flake-graphene oxide nanocomposite.
[0045] 3. Method:
[0046] (1) AC 60 / ITO: put 10 μL of AC 60 Solution (1mg·mL -1 ) was added dropwise to the surface of the cleaned ITO electrode, and after drying, the electrochemical workstation was used to detect AC under light. 60 Photocurrent-potential curve of / ITO, photoelectrochemical tests were performed with 10 mM phosphate buffer solution containing 0.1 M ascorbic acid as electrolyte and -0.2 V as bias.
[0047] (2) AC 60 -GO / ITO: put 10 μ...
Embodiment 2
[0064] Example 2: Incubation of Abs 2 @AC 60 -Photocurrent-potential maps before and after Gr-GO
[0065] 1. Instrument: Shanghai Chenhua Electrochemical Workstation (chi 600e software); 150 watt xenon lamp
[0066] 2. Materials: indium tin oxide transparent conductive glass electrode (ITO); GO dispersion; capture antibody (alpha-fetoprotein primary antibody, Ab 1 ); antigen to be tested (alpha-fetoprotein, AFP); labeled antibody (secondary antibody to alpha-fetoprotein, Ab 2 ); 1 wt.% aqueous bovine serum albumin
[0067] 3. Method:
[0068] (1) Add 10 μL (0.1 mg·mL) dropwise to the surface of the cleaned ITO electrode -1 ) GO dispersion solution, then drop 10 μL (100mM / 50mM) EDC / NHS solution on the electrode surface, and place it at 37°C for 2h to activate the carboxyl groups;
[0069] (2) Drop 10 μL (10 μg·mL) on the surface of the electrode obtained in step (1). -1 )Ab 1 solution, placed at 4°C for 12h to make Ab 1 Covalently binds to GO and is fully linked;
[0...
Embodiment 3
[0079] Example 3: Ab 2 Preparation of @AC60-Gr-GO
[0080] (a) 1 mg AC 60 , 1mg Gr, 1mg GO were placed in an agate mortar, and 100 μL of secondary water was added to grind for 1 h;
[0081] (b) Disperse the ground material into 10 mL of water, and ultrasonically treat it for 4 h;
[0082] (c) Centrifuge at 3000rpm for 30min, collect the precipitate, and redisperse it in 5mL of water to obtain AC 60 -Gr-GO dispersion.
[0083] (d) Take 200 μL AC 60 -Add 10μL of freshly prepared EDC / NHS solution (100mM / 50mM) dropwise to the Gr-GO dispersion, sonicate at 20°C for 30min, and then add 200μL of 100μg·mL -1 Ab 2 The solution was stirred at 4°C for 12h, added with 2mg BSA, stirred at room temperature for 1h, then centrifuged and resuspended in 0.4mL PBS solution to obtain Ab 2 @AC 60 -Gr-GO solution.
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