Biosensor and application thereof
A technology of biosensors and markers, applied in the field of biosensors, can solve problems such as the inability to realize multi-indicator joint detection, and achieve the effects of low preprocessing requirements, large detection throughput, and high degree of automation
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Embodiment 1
[0049] The preparation of embodiment 1 colloidal gold
[0050] After diluting 1.0% (w / v) chloroauric acid aqueous solution with pure water 100 times, measure 500mL of 0.01% (w / v) chloroauric acid aqueous solution in a 1L beaker, and heat it to 95°C while electromagnetic stirring , continued for 10 minutes, then added 5 mL of 1.0% (w / v) trisodium citrate aqueous solution, boiled for more than 15 minutes, until the color of the solution turned orange red, and cooled at room temperature to obtain a colloidal gold solution, which was stored at 4°C for later use.
Embodiment 2
[0051] Preparation of Example 2 Colloidal Gold Labeled Monoclonal Antibody
[0052] The colloidal gold labeling method of AFP monoclonal antibody is as follows:
[0053] Take 10mL colloidal gold solution, use 1.0mol / L potassium carbonate (K 2 CO 3 ) aqueous solution to adjust the pH to 7.4, and stir magnetically for 5 minutes at room temperature; add 10 μL of 2 mg / mL AFP antibody, use HCl to adjust the pH to an acidic reaction environment suitable for the antibody, and stir at room temperature for 15 minutes; then add 1 mL of 10% BSA and continue stirring Cool to 4°C, adjust the pH to 7.4 with NaOH; centrifuge at 10000r / min at 4°C for 10min, add 100μL PBS (10mmol / L, pH7.4) to the pellet to resuspend the particles, and obtain colloidal gold-labeled AFP monoclonal Antibody solution; finally concentrated by desalting column and diluted with PBS (10mmol / L, pH7.4) to obtain 2mg / mL colloidal gold-labeled AFP monoclonal antibody.
[0054] Using the same method, the AFP monoclonal ...
Embodiment 3
[0055] Monoclonal antibody modification of embodiment 3 biosensor probe
[0056] (1) Carboxylation modification of biosensor probes
[0057] Take 6.6mL of APTES (3-aminopropyltriethoxysilane, 3-aminopropyltriethoxysilane) and dissolve it in 50mL of absolute ethanol to prepare a 0.6M APTES ethanol solution, seal it with a cover and store it at 4°C for later use;
[0058] Dissolve 0.15g of succinic anhydride in 50mL of N,N-dimethylformamide (DMF) to prepare an anhydride solution, and store it at 4°C for later use;
[0059] Sonicate the probe in pure water for 10 min, in absolute ethanol for 10 min, in the treatment solution (35 mL of concentrated sulfuric acid and 15 mL of hydrogen peroxide) for 30 min, in absolute ethanol for 10 min, and blow dry with nitrogen;
[0060] Immerse the treated clean and dry probe into 0.6M APTES ethanol solution for about 5mm, and keep it at 50°C for more than 12 hours; about 5mm in absolute ethanol for 10min; about 5mm in acid anhydride solution ...
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