Construction method and application of polydopamine-mediated modification-free portable conductivity immunosensor with adjustable linear range
An immune sensor and polydopamine technology, applied in the field of analysis and detection, can solve problems such as poor stability and reproducibility, and achieve the effect of improving field operability, simple operation, and good stability
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Embodiment 1
[0062] Example 1 Modification of magnetic beads with biological recognition molecules
[0063] 1. Activation of magnetic beads
[0064] 1) Take 2 mg of magnetic beads (average diameter 1 μm) in a centrifuge tube, wash twice with 500 μL MEST (10 mM MES, 0.05% Tween20, pH 6.0), and remove the supernatant by magnetic separation;
[0065] 2) Prepare 5mg / mL EDC solution and 5mg / mL NHS solution with 10mM MES (pH 6.0);
[0066] 3) Add 100 μL EDC (5 mg / mL) and 50 μL NHS (5 mg / mL) to the centrifuge tube containing magnetic beads, mix well with a vortexer to fully suspend the magnetic beads, dilute to 500 μL with MES, and place in a On a rotary mixer, activate at 37°C for 30min;
[0067] 4) Magnetic separation, remove the supernatant, add 500 μL of MEST, and transfer the magnetic beads to a new centrifuge tube;
[0068] 5) Magnetic separation, remove the supernatant, wash twice with 500 μL of MEST, magnetic separation, remove the supernatant.
[0069] After the above steps, the carb...
Embodiment 2
[0077] Example 2 Synthesis and Biomodification of Gold Core Platinum Shell Nanometers
[0078] 1. Synthesis of gold core platinum shell nanozyme
[0079] In order to avoid the impurity of the prepared nanoparticles, the utensils required for the experiment should be soaked in freshly prepared aqua regia (concentrated nitric acid:concentrated hydrochloric acid: 1:3) for 3h before the experiment, rinsed with ultrapure water, and dried for use. use. The preparation of AuNPs introduced the microwave synthesis method, and the preparation of AuNPs could be completed in 5 minutes. Add 200 μL HAuCl4 (48 μM) and 9.8 mL ultrapure water to the reaction kettle, and keep boiling for 1 min under microwave heating at 100 °C. Then, the magnetic stirring was turned on, and 1 mL of 1 wt% sodium citrate was rapidly added at one time, kept at 100 °C for 4 min, and then cooled to room temperature to obtain a red AuNPs suspension. Au@PtNPs were synthesized by platinum precipitation on the surface ...
Embodiment 3
[0085] Example 3 Optimization of reaction conditions for conductivity immunosensors without signal amplification strategy
[0086] Different mass (20 μg, 50 μg, 100 μg, 150 μg) of DON complete antigen was coupled to the magnetic beads, and other experimental steps were as described above. The result is as figure 2 As shown, the conductivity immunosensor has the best response effect when the complete antigen concentration is 20 μg.
[0087] Different concentrations (1 μg / mL, 2 μg / mL, 3 μg / mL, 4 μg / mL) of DON monoclonal antibody were used for competitive immune reaction, and other experimental steps were as described above. The result is as Figure 4 As shown, the conductivity immunosensor has the best response effect when the concentration of spittoxin monoclonal antibody is 2 μg / mL.
[0088] Dopamine hydrochloride solutions with different concentrations (1mg / mL, 3mg / mL, 5mg / mL, 10mg / mL) were set up, and other experimental steps were as described above. The result is as ...
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