Acidity regulation and lectin recognition Escherichia coli visualization biosensing method
A technology of Escherichia coli and biosensing, which is applied in the field of nanomaterial science and biosensing, and achieves the effects of broad application prospects, simple modification operation, convenient and sensitive detection
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Embodiment 1
[0045] Add 47 mL of ultrapure water, 10 mg of dextran, and 0.6 mL of HAuCl to a clean glass vessel 4 (98mM), ultrasonically dispersed to obtain a uniformly dispersed clear solution; add 1mL NaOH (1M) to the above mixed solution under rapid stirring, adjust the pH to 11, keep stirring for 12-16 hours, when the color of the solution turns wine red Stop the reaction, transfer to a clean glass bottle, and store at 4°C. The dextran-modified gold nanoparticles were centrifuged at 1000 rpm for 5 minutes, dispersed with phosphate buffer (20 mM, pH 6.8), and stored at low temperature for future use.
[0046] Put 15mL of magnetic bead aqueous solution in a 50mL centrifuge tube, and disperse for 5min under the action of ultrasound with a working frequency of 40kHz and a power of 160W to obtain a uniformly dispersed magnetic bead dispersion. Perform magnetic separation and remove the supernatant in sequence, wash the magnetic beads three times with phosphate buffer, then disperse in 15 m...
Embodiment 2
[0059] Put 15mL of magnetic bead aqueous solution in a 50mL centrifuge tube, and disperse for 5min under the action of ultrasound with a working frequency of 40kHz and a power of 160W to obtain a uniformly dispersed magnetic bead dispersion. Perform magnetic separation, remove supernatant, and wash magnetic beads three times with phosphate buffer, then disperse in 15 mL of MES buffer (final concentration of magnetic beads is about 1 mg / mL), and store at 4°C.
[0060] Add EDC (2 mg) and NHS (0.35 mg) to 1 mL of the above-mentioned functional magnetic bead dispersion, stir and react at room temperature for 2 hours, wash with phosphate buffer (pH 6.0) magnetically for three times, and then disperse in 1 mL of phosphate buffer , to obtain the activated magnetic bead dispersion. Take the above-mentioned activated magnetic beads and 0.2mL streptavidin (1mg / mL) to react at room temperature for 1 hour, use phosphate buffer (pH 6.0) to magnetically separate, remove the supernatant, and...
Embodiment 3
[0066] Add 47 mL of ultrapure water, 10 mg of dextran, and 0.6 mL of HAuCl to a clean glass vessel 4 (98mM), ultrasonically dispersed to obtain a uniformly dispersed clear solution; add 1mL NaOH (1M) to the above mixed solution under rapid stirring, adjust the pH to 11, keep stirring for 12-16 hours, when the color of the solution turns wine red Stop the reaction, transfer to a clean glass bottle, and store at 4°C.
[0067] The dextran-modified gold nanoparticles were centrifuged at 1000 rpm for 5 minutes, dispersed with phosphate buffer (20 mM, pH 6.8), and stored at low temperature for future use. Take 100 μL dextran-modified gold nanoparticles and the lectin ConA with final concentrations of 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 μg / mL for mixed reaction, and after incubation for 1 min, Take the nano-gold mixed solution into a micro-volume fluorescence cuvette, and measure the absorption spectrum at 400nm-700nm with an ultraviolet spectrophotometer.
[0068] The UV-Vis abso...
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