Biological probe and biological immunosensor for detecting bungarus multicinctus venom and method for detecting bungarus multicinctus venom based on proportional signal
An immune sensor and Bungare venom technology, which is applied in biological testing, biological material analysis, instruments, etc., can solve the problems of long detection time and high equipment cost, and achieve the effects of short detection time, high sensitivity and wide repeatability
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[0042] In the present invention, the preparation method of the detection bioprobe based on the ratio-type double signal of the bungalow snake venom comprises the following steps:
[0043] (1) Mix 1,3,5-tri(4-aminophenyl)benzene, 2,5-dimethoxybenzene-1,4-dicarbaldehyde, acetic acid with an organic solvent, and carry out solvothermal reaction to obtain a total Valence organic framework materials;
[0044] (2) the covalent organic framework material and alcohol solvent, HAuCl 4and reducing agent are mixed for reduction reaction to obtain AuNPs / COF nanocomposite material;
[0045] (3) Mixing the AuNPs / COF nanocomposite material with a sulfhydryl-modified bungalo β toxin aptamer to obtain a bioprobe for detecting bungale venom based on a ratiometric dual signal.
[0046] The present invention mixes 1,3,5-tris(4-aminophenyl)benzene, 2,5-dimethoxybenzene-1,4-dicarbaldehyde, acetic acid and an organic solvent for solvothermal reaction to obtain covalent Organic Frameworks (COFs). ...
Embodiment 1
[0105] A preparation method of a proportional chemical bioimmunosensor for detecting bununga venom, the specific steps are as follows:
[0106] (1) Preparation of AuNPs@COF: 0.030 mmol of 1,3,5-tris(4-aminophenyl)benzene and 0.030 mmol of 2,5-dimethoxybenzene-1,4-dicarbaldehyde were added together 4.5 mL of 1,4-dioxane-butanol-methanol (V 1 / V 2 / V 3 ,4:4:1) co-solvent system. Then, the mixture was sonicated for 20 min to obtain a homogeneous mixed solution. After adding 0.050 mL of 12M acetic acid aqueous solution dropwise, let stand at room temperature for 2 hours, then add 0.45 mL of 12M acetic acid aqueous solution, and heat at 70°C for 24 hours. Finally, the product was cooled to room temperature, and the synthesized COF was washed three times with THF and acetone, and dried in vacuum for 24 h. The obtained COF (0.060g) was added to 30mL of methanol, hydrochloric acid was added to adjust the pH to 4, and then 200 μL of 1% HAuCl was slowly added dropwise 4 4H 2 O in...
Embodiment 2
[0118] A preparation method of a proportional chemical bioimmunosensor for detecting bununga venom, the specific steps are as follows:
[0119] (1) Preparation of AuNPs@COF: 0.030 mmol of 1,3,5-tris(4-aminophenyl)benzene and 0.040 mmol of 2,5-dimethoxybenzene-1,4-dicarbaldehyde were added together 4.5 mL of 1,4-dioxane-butanol-methanol (V1 / V2 / V3, 4:4:1) co-solvent system. Then, the mixture was sonicated for 20 min to obtain a homogeneous mixed solution. After adding 0.050 mL of 12M acetic acid aqueous solution dropwise, let stand at room temperature for 2 hours, then add 0.45 mL of 12M acetic acid aqueous solution, and heat at 70°C for 24 hours. Finally, the product was cooled to room temperature, and the synthesized COF was washed three times with THF and acetone, and dried in vacuum for 24 h. Add the obtained COF (0.060g) into 30mL of methanol, add hydrochloric acid to adjust the pH to 4, then slowly add 200μL of 1% HAuCl4·4H2O into the mixed solution, stir at 0°C for 5h, ...
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