An electrochemical immunosensor for detecting epidermal growth factor and its preparation method
An epidermal growth factor and immunosensor technology, applied in the field of electrochemical immunosensors and their preparation, can solve the problems of uneven quality of ESLIA kits, inability to detect low-content substances, and organic solvents that are harmful to the human body, and achieve excellent stability, The effect of rapid detection and improved sensitivity
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Embodiment 1
[0054] Embodiment 1, preparation of polydopamine viscous layer, using the functionalized nano-silver obtained in embodiment 1 to recognize and detect EGF
[0055] 1. Preparation of nano-silver sol
[0056] 10ml1×10 -3 MAgNO 3 The aqueous solution was added dropwise to 40ml of 2×10 in an ice-water bath stirred with a magnetic stirrer -3 MNbH 4 In the aqueous solution, to be AgNO 3 After the aqueous solution was added dropwise, the magnetic stirring reaction was continued for 10 minutes to obtain nano-silver sol, wherein the nano-silver was bright yellow and transparent.
[0057] 2. Preparation of nano-silver sol modified by antibody to epidermal growth factor
[0058] 1) Take 3 μl of thioglycolic acid adjusted to pH to neutral with KOH concentrated solution, add it to 5ml nano-silver sol, and magnetically stir for 15 minutes to obtain thioglycolic acid-modified nano-silver sol;
[0059] 2) The pH of the nano-silver sol modified by thioglycolic acid was adjusted to 5-6, an...
Embodiment 2
[0080] According to the steps of Example 1, AgNO will be added 3 The continuous magnetic stirring time after the solution was replaced with 5min; the continuous magnetic stirring time after adding thioglycolic acid was replaced with 10min; the magnetic stirring time was replaced with 10min after adding EDC and NHS solution; the shaker oscillation time was replaced with 1hour.
[0081] In the embodiment 1 step, NaBH 4 The solution concentration is replaced by 1×10 -3 mol / L; AgNO 3 The solution concentration is replaced by 0.5×10 -3 mol / L; the concentration of EDC solution is replaced by 1×10 -3 mol / L, the concentration of NHS solution is replaced by 1×10 -3 mol / L,; the volume of the epidermal growth factor antibody was replaced by 10 μl; in the cyclic voltammetry detection, the potential sweep rate was 50mV﹒ S -1 .
[0082] In the step of embodiment 1, the electrode is immersed in dopamine tris buffer solution, and the soaking time is replaced by 20min, and the electrode...
Embodiment 3
[0085] According to the steps of Example 1, AgNO will be added 3 The continuous magnetic stirring time after the solution was replaced with 20min; the continuous magnetic stirring time after adding thioglycolic acid was replaced with 30min; the magnetic stirring time was replaced with 30min after adding EDC and NHS solution; the shaker oscillation time was replaced with 4hour.
[0086] In the embodiment 1 step, NaBH 4 The solution concentration is replaced by 5×10 -3 mol / L; AgNO 3 The solution concentration is replaced by 3×10 -3 mol / L; the concentration of EDC solution is replaced by 5×10 -3 mol / L, the concentration of NHS solution is replaced by 5×10 -3 mol / L; the volume of the epidermal growth factor antibody was replaced with 40 μl; in the cyclic voltammetry detection, the potential sweep rate was 200mV﹒ S -1 .
[0087] In the step of embodiment 1, the electrode is immersed in dopamine tris buffer solution, and the soaking time is replaced by 40min, and the electrod...
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