Construction of high-throughput electrochemiluminescence detection method for early tumor
A luminescence detection and electrochemical technology, applied in chemiluminescence/bioluminescence, analysis by chemical reaction of materials, measurement devices, etc. problem, to achieve the effect of strong designability, low price and low toxicity
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Embodiment 1
[0018] Example One Combination figure 1 , figure 2 and image 3 , Preparation of iridium complex-filled mesoporous silicon / nano-gold composite signaling probes
[0019] 1.0mL concentration is 2.5 × 10 −4 M [(ppy) 2 Ir(dcbpy)] + PF 6 − ( figure 1 ) was added to the same volume of 1.0 mg / mL mesoporous silicon solution and stirred overnight at 37°C, then 1.5 mL of 3-mercaptopropyltrimethoxysilane was added and stirred for 2 hours, then the nano-gold solution was added. Nanogold is assembled onto the surface of mesoporous silicon through the combination of gold-mercapto covalent bonds to form a stable iridium complex-filled mesoporous silicon / nanogold complex signal probe. Take 1mL of the above nanocomposite and add 1mL of 0.02mg / mL antibody (secondary antibody, 0.01M PBS, pH=7.4) was incubated at room temperature for 2 h. Centrifuge, wash with 0.0 1M PBS with pH=7.4, then dissolve the resultant in 1 mL of 1% BSA, incubate for 1 h, and wash by centrifugation. The fina...
Embodiment 2
[0020] Example two combination Figure 4 , the development of flexible array microchip electrodes
[0021] Mix polydimethylsiloxane (PDMS) with an appropriate amount of curing agent at a mass ratio of 9:1, and form a transparent electrode base material after curing. Make a hollow plexiglass plate (the hollow part is in the shape of an electrode), and cover it tightly on the PDMS sheet. will contain 1% HAuCl 4 , 200 g L -1 KHCO 3 and 2% glucose (volume ratio: 2:1:1) electroless gold-plating solution was injected into the above hollowed out part, after the air bubbles were exhausted, it was left to stand for 3 hours under dark conditions, and then the gold-plating solution was removed to form a layer of texture on the surface of PDMS Uniform gold nanofilms. The nano-gold layer has excellent electrical conductivity. At the same time, with the flexibility of the substrate PDMS material, the obtained flexible microchip electrode has unique advantages in preparation, storage, ...
Embodiment 3
[0022] Example three combination Figure 5 , High-sensitivity, high-throughput electrochemiluminescent tumor marker detection based on novel signaling probes
[0023] On the surface of the designed flexible microchip, the bionic sensing interface is constructed by using the interface modification technology. Add 10 μL of primary antibody (100 μg L -1 , 0.01 M pH 7.4 PBS) onto the surface of the electrode and incubated at 4 °C for 12 h, and the unbound antibody was removed by washing with buffer. Then the prepared electrode was immersed in 1 % (w / w) BSA solution for 1 h at room temperature to seal the possible active sites and non-specific adsorption on the electrode surface, and finally the electrode was washed several times with buffer solution. The modified electrodes were stored in a refrigerator at 4 °C until use. Soak the prepared immune electrode in 100 μL of tumor marker solutions of different concentrations, incubate at 37 °C for 40 min, take it out and wash it seve...
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