An electrochemical sensor for detecting ache inhibitors
An electrochemical and sensor technology, applied in the field of biosensors, can solve the problems of low specificity and sensitivity, high cost, and achieve the effects of simple preparation method, simple electrode and mild reaction conditions
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Embodiment 1
[0038] Example 1 DNAzyme Walker concentration screening
[0039] (1) The gold electrode was polished in 0.3 and 0.05 µM alumina slurry in turn until it became a mirror surface, and then PBS buffer and ddH were used in turn. 2 O were washed 3-5 times repeatedly;
[0040] (2) DNAzyme solution or ddH concentration of 25 nM, 50 nM, 75 nM, 100 nM respectively 2 10 µL each of O and 1 µM HP chain solution were added dropwise to the surface of the gold electrode obtained in step (1), incubated at 37 °C for 2 h, ddH 2 O rinse 3 times;
[0041] (3) Put 10 μL of Hg(NO) at a concentration of 150 nM 3 ) 2 The solution was added dropwise to the surface of the gold electrode obtained in step (1), incubated at 37 °C for 2 h to block DNAzyme activity, ddH 2 O rinse 3 times;
[0042] (4) The gold electrode obtained in step (3) was prepared in PBS buffer (pH 6.2, 2.5 mM Mg) containing 2 μM STP. 2+ ) at 25 °C for 2 h;
[0043] (5) Mix 4 μL of 30 μM acetylcholine and 4 μL of 1 μM acetylcho...
Embodiment 2
[0046] Example 2 HP chain concentration screening
[0047] (1) The gold electrode was polished in 0.3 and 0.05 µM alumina slurry in turn until it became a mirror surface, and then PBS buffer and ddH were used in turn. 2 O were washed 3-5 times repeatedly;
[0048] (2) The DNAzyme solution with a concentration of 50 nM was mixed with 0.5 μM, 1 μM, 1.5 μM, 2 μM HP chain solution or ddH, respectively. 2 10 µL of O was added dropwise to the surface of the gold electrode obtained in step (1), incubated at 37 °C for 2 h, ddH 2 O rinse 3 times;
[0049] (3) Put 10 μL of Hg(NO) at a concentration of 150 nM 3 ) 2 The solution was added dropwise to the surface of the gold electrode obtained in step (1), incubated at 37 °C for 2 h to block DNAzyme activity, ddH 2 O rinse 3 times;
[0050] (4) The gold electrode obtained in step (3) was prepared in PBS buffer (pH 6.2, 2.5 mM Mg) containing 2 μM STP. 2+ ) at 25 °C for 2 h to obtain a detection electrode;
[0051] (5) Mix 4 μL of 30...
Embodiment 3
[0054] Example 3 Screening of reaction time
[0055] (1) The gold electrode was polished in 0.3 and 0.05 µM alumina slurry in turn until it became a mirror surface, and then PBS buffer and ddH were used in turn. 2 O were washed 3-5 times repeatedly;
[0056] (2) 10 μL of DNAzyme solution with a concentration of 50 nM and 10 μL of 1 μM HP chain solution were added dropwise to the surface of the gold electrode obtained in step (1), and incubated at 37 °C for 2 h. 2 O rinse 3 times;
[0057] (3) Put 10 μL of Hg(NO) at a concentration of 150 nM 3 ) 2 The solution was added dropwise to the surface of the gold electrode obtained in step (1), incubated at 37 °C for 2 h to block DNAzyme activity, ddH 2 O rinse 3 times;
[0058] (4) The gold electrode obtained in step (3) was prepared in PBS buffer (pH 6.2, 2.5 mM Mg) containing 2 μM STP. 2+ ) at 25 °C for 2 h to obtain a detection electrode;
[0059] (5) Mix 4 μL of 30 μM acetylcholine and 4 μL of 1 μM acetylcholinesterase with...
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