Electrochemical sensor for detecting AChE inhibitor
An electrochemical and sensor technology, applied in the field of biosensors, can solve the problems of high cost, low specificity and sensitivity, and achieve the effects of simple electrodes, simple preparation methods, and sensitive detection.
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Embodiment 1
[0038] Example 1 DNAzyme Walker Concentration Screening
[0039] (1) The gold electrode is polished in 0.3 and 0.05 µM alumina slurry in turn until it becomes a mirror surface, and then it is polished with PBS buffer and ddH 2 O wash repeatedly 3-5 times respectively;
[0040] (2) DNAzyme solution or ddH with concentrations of 25 nM, 50 nM, 75 nM and 100 nM respectively 2 10 µL each of O and 1 μM HP chain solution was dropped onto the surface of the gold electrode obtained in step (1), and incubated at 37 °C for 2 h, ddH 2 O wash 3 times;
[0041] (3) Add 10 μL of 150 nM Hg(NO 3 ) 2 The solution was added dropwise to the surface of the gold electrode obtained in step (1), and incubated at 37 °C for 2 h to block DNAzyme activity, ddH 2 O wash 3 times;
[0042] (4) The gold electrode obtained in step (3) was placed in PBS buffer (pH 6.2, 2.5 mM Mg 2+ ) at 25°C for 2 h;
[0043] (5) Mix 4 μL of 30 μM acetylcholine and 4 μL of 1 μM acetylcholinesterase with 2 μL of Tris-HC...
Embodiment 2
[0046] Example 2 HP chain concentration screening
[0047] (1) The gold electrode is polished in 0.3 and 0.05 µM alumina slurry in turn until it becomes a mirror surface, and then it is polished with PBS buffer and ddH 2 O wash repeatedly 3-5 times respectively;
[0048] (2) Mix DNAzyme solution with a concentration of 50 nM with 0.5 μM, 1 μM, 1.5 μM, 2 μM HP chain solution or ddH 2 10 µL each of O was dropped onto the surface of the gold electrode obtained in step (1), and incubated at 37 °C for 2 h, ddH 2 O wash 3 times;
[0049] (3) Add 10 μL of 150 nM Hg(NO 3 ) 2 The solution was added dropwise to the surface of the gold electrode obtained in step (1), and incubated at 37 °C for 2 h to block DNAzyme activity, ddH 2 O wash 3 times;
[0050] (4) The gold electrode obtained in step (3) was placed in PBS buffer (pH 6.2, 2.5 mM Mg 2+ ) at 25°C for 2 h to obtain the detection electrode;
[0051] (5) Mix 4 μL of 30 μM acetylcholine and 4 μL of 1 μM acetylcholinesterase wi...
Embodiment 3
[0054] Example 3 Screening of reaction time
[0055] (1) The gold electrode is polished in 0.3 and 0.05 µM alumina slurry in turn until it becomes a mirror surface, and then it is polished with PBS buffer and ddH 2 O wash repeatedly 3-5 times respectively;
[0056] (2) Add 10 µL each of DNAzyme solution with a concentration of 50 nM and 1 μM HP chain solution onto the surface of the gold electrode obtained in step (1), incubate at 37 °C for 2 h, ddH 2 O wash 3 times;
[0057] (3) Add 10 μL of 150 nM Hg(NO 3 ) 2 The solution was added dropwise to the surface of the gold electrode obtained in step (1), and incubated at 37 °C for 2 h to block DNAzyme activity, ddH 2 O wash 3 times;
[0058] (4) The gold electrode obtained in step (3) was placed in PBS buffer (pH 6.2, 2.5 mM Mg 2+ ) at 25°C for 2 h to obtain the detection electrode;
[0059] (5) Mix 4 μL of 30 μM acetylcholine and 4 μL of 1 μM acetylcholinesterase with 2 μL of Tris-HCl buffer solution (10 mM Tris, 10 mM Mg ...
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