Electrochemical luminescence immune sensor built by mesoporous material loading quantum dots and coated by nanogold, and detection method of HIV (human immunodeficiency virus)
A luminescence immunity, sensor technology, applied in chemiluminescence/bioluminescence, analysis by chemical reaction of materials, scientific instruments, etc., can solve the problems of low background signal of excitation spectrum, narrow emission spectrum, etc., to achieve wide application prospects, High sensitivity, simple detection effect
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Embodiment 1
[0024] Example 1: Preparation of mesoporous silica loaded with quantum dots and coated with gold nanoparticles
[0025] Quantum dots used in the present invention, mesoporous silicon oxide, and nano-gold are respectively synthesized according to the method reported in the literature, and the specific method is referred to in the literature (Talanta, 80, 1737-1743 (2010), Langmuir, 27, 6099-6106 (2011), Anal .Chem., 77,3324-3329(2005).The specific preparation scheme of the mesoporous silica loaded with quantum dots and coated with nano gold is as follows:
[0026] (1) Disperse 50mg of mesoporous silica in 10mL of anhydrous toluene, add the same volume of water-soluble CdTe quantum dots and stir evenly at 40°C for 24 hours, centrifuge, wash with distilled water and absolute ethanol at least three times each, vacuum Dry to obtain mercapto-modified mesoporous silica.
[0027] (2) Disperse 20 mg of mercapto-modified mesoporous silica in 10 mL of tetrahydrofuran, add the same volum...
Embodiment 2
[0029] Example 2: Construction of Electrochemiluminescence Immunosensor
[0030] The present invention constructs an electrochemiluminescent immunosensor by using the above-mentioned mesoporous silicon oxide loaded with quantum dots and coated with nano-gold on the surface together with magnetic beads, and the specific embodiments are as follows:
[0031] (1) Preparation of magnetic capture probe: 10mg mL -1 Ultrasonic dispersion of magnetic beads in PBS buffer at pH 7.4, pipette 1mL, 50μg mL -1 Goat anti-human anti-antibody anti-IgG was added, mixed and stirred at 4°C for 6 hours, and the active sites of other unbound antibodies were blocked with 1 mL of 1% bovine serum albumin (BSA), and the anti-anti-body that was not fixed by magnetic beads was washed by magnetic separation. -IgG, after washing, redisperse in 1mL PBS solution of pH7.4 to obtain a magnetic capture probe (NMP / anti-IgG); put the magnetic capture probe (NMP / anti-IgG) prepared above into the solution Add a ce...
Embodiment 3
[0034] Example 3 Electrochemiluminescence response of several different modified electrodes
[0035] Using different materials to modify the electrode, compare its electrochemiluminescent response, the results are as follows figure 1 shown. bare electrode ( figure 1 , curve a) contains 0.1mol / L PBS (pH=7.4) and 0.1mol / L K in 0.1mol / L KCl 2 S 2 o 8 Electrochemiluminescence behavior in solution, the electrochemiluminescence response signal is very weak; when the electrode surface modified NMP / anti-IgG ( figure 1 , after curve b), its electrochemiluminescent signal is also very weak, this is because the electrode surface is covered with a layer of insulating protein layer, and there is no electrochemiluminescent source material, so its electrochemiluminescent signal is almost non-existent; the same Principle, when the electrode surface is modified with NMP / anti-IgG / Ab( figure 1 , after curve c), the electrochemiluminescence signal also remains unchanged; when the electrode ...
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