An electrochemiluminescent immunosensor for detecting circulating tumor cells and its preparation method and application
A technology of tumor cells and electrochemistry, applied in the field of biomedical detection, to achieve the effect of high selectivity and excellent anti-interference performance
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Embodiment 1
[0048] Embodiment 1 Magnetic ECL probe Fe 3 o 4 @SiO 2 / AuNPs / WS 2 - Preparation of Aptamer
[0049] (1) Fe 3 o 4 @SiO 2 preparation of
[0050] Weigh 0.65g of ferric chloride and 0.2g of trisodium citrate and add them to 20ml of ethylene glycol, put them on a magnetic stirrer and stir to dissolve, then add 1.2g of sodium acetate while stirring, stir for 30min, and turn to Put it into a high-temperature and high-pressure reactor at 200 ° C, react for 10 hours, take it out after the reaction and cool it to room temperature, and centrifuge and wash to obtain Fe 3 o 4 Nanoparticles. 0.1g prepared Fe 3 o 4 Dissolve in ethanol-water mixture (30mL, volume ratio 2:1), add 0.1g ammonia water, 0.1g formaldehyde solution (37%) and 0.1 resorcinol to it, stir mechanically at 30°C for 10h, then heat up to Fe can be obtained at 100°C 3 o 4 composite nanomaterials. Dissolve 0.2g of the material prepared above in 80mL aqueous solution containing 0.5g CTAB and 0.8mL ammonia wate...
Embodiment 2
[0055] Example 2 Assembly of Electrochemiluminescence Immunosensor (ECL Immunosensor) and Its Measurement Method
[0056] The detection principle of the electrochemiluminescence immunosensor is as follows: figure 1 shown, including the following steps:
[0057] (1) Electrode pretreatment: use 0.05 and 0.03 μm Al on glassy carbon electrodes respectively 2 o 3 Powder treatment, and then ultrasonic cleaning with absolute ethanol and ultrapure water for 5 minutes;
[0058] (2) Modification of the base electrode: Take 5 μL of 0.5% Nafion / AuNPs solution with a pipette gun and drop it on the surface of the pretreated glassy carbon electrode, leave it at room temperature for 2 hours to dry, wash the unbound composite material with PBS solution, dry it, and use it to The advantage of the larger specific surface area makes more antibodies enriched on the surface of the electrode; the electrode is soaked in a concentration of 1mM Ru(bpy) 3 2+ solution, placed at room temperature for...
Embodiment 3
[0064] Example 3 Determination of the ECL immunosensor cyclic voltammogram of MCF-7 cells
[0065] In order to explore the signal response of the ECL immunosensor of MCF-7 cells at each modification stage, the glassy carbon electrode obtained in each step in Example 2 was placed in a 0.01M PBS solution for cyclic voltammetry scanning at a speed of 0.1V / s , see the result figure 2 shown. With Nafion / AuNPs / Ru(bpy) 3 2+ Modified on the surface of the glassy carbon electrode, an increased peak current signal was obtained compared with the bare electrode, because Ru(bpy) 3 2+Enhanced electron transfer improves sensor sensitivity. Subsequently, antibodies, BSA, and MCF-7 cells with labeled probes were modified on the electrode surface, and the response signal of the sensor gradually decreased. The reason was that the protein increased the impedance value of the electrode surface and reduced the current.
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