An electrochemiluminescent biosensor for detecting mmp-14 and its preparation method
A technology of MMP-14 and luminescent organisms, which is applied in the field of medical devices, can solve the problems of the influence of sensor detection sensitivity, the difficulty of applying detection of MMPs, and the lack of specificity of biosensing technology, so as to achieve the effect of reducing non-specific adsorption
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[0038] A method for preparing an electrochemiluminescence biosensor for detecting MMP-14, the specific preparation steps are as follows:
[0039] (1) fullerene-chitosan (C 60 -Chit) preparation
[0040] First configure 0.2-2mg / mL of C 60 Solution, ultrasonic 20-30min, guarantee C 60Uniformly dispersed in the solution, and then configure a chitosan solution with a mass concentration of 0.5%. Mix the two solutions in equal volume and stir for 40-60min to ensure that C 60 Uniformly dispersed in chitosan solution, that is, C 60 -Chit.
[0041] (2) Preparation of MMP-14 protein-specific recognition probe
[0042] The polypeptide inhibitor Ap1 was dissolved in Tris-HCl (pH 8.0) buffer, and the concentration of Ap1 was 3-50 μM.
[0043] (3) Assembly of the electrochemiluminescence sensor
[0044] a. Polish the glassy carbon electrode with a diameter of 3mm with 1.0μM, 0.3μM, 0.05μM Al2O3 powder in sequence, then clean it with ethanol and deionized water in sequence, and then ...
specific Embodiment 1
[0058] A method for preparing an electrochemiluminescence biosensor for detecting MMP-14, the specific preparation steps are as follows:
[0059] (1) fullerene-chitosan (C 60 -Chit) preparation
[0060] First configure 0.2mg / mL of C 60 Solution, ultrasonic 20min, guarantee C 60 Uniformly dispersed in the solution, and then configure a chitosan solution with a mass concentration of 0.5%. Mix the two solutions in equal volumes and stir for 40 min to ensure that C 60 Uniformly dispersed in chitosan solution, that is, C 60 -Chit.
[0061] (2) Preparation of MMP-14 specific recognition signal probe
[0062] The polypeptide inhibitor Ap1 was dissolved in Tris-HCl (pH 8.0) buffer, and the concentration of Ap1 was 3 μM.
[0063] (3) Assembly of the electrochemiluminescence sensor
[0064] a. Polish the glassy carbon electrode with a diameter of 3mm with 1.0μM, 0.3μM, and 0.05μM Al2O3 powder in sequence, then ultrasonically clean it with ethanol and deionized water in turn, and...
specific Embodiment 2
[0069] A method for preparing an electrochemiluminescence biosensor for detecting MMP-14, the specific preparation steps are as follows:
[0070] (1) fullerene-chitosan (C 60 -Chit) preparation
[0071] First configure 1mg / mL of C 60 Solution, ultrasonic 25min, guarantee C 60 Uniformly dispersed in the solution, and then configure a chitosan solution with a mass concentration of 0.5%. Mix the two solutions in equal volumes and stir for 45 min to ensure that C 60 Uniformly dispersed in chitosan solution, that is, C 60 -Chit.
[0072] (2) Preparation of MMP-14 specific recognition signal probe
[0073] The polypeptide inhibitor Ap1 was dissolved in Tris-HCl (pH 8.0) buffer, and the concentration of Ap1 was 9 μM.
[0074] (3) Assembly of the electrochemiluminescence sensor
[0075] a. The glassy carbon electrode with a diameter of 3mm is polished with 1.0μM, 0.3μM, 0.05μM Al2O3 powder in turn, and then ultrasonically cleaned with ethanol and deionized water in turn, and t...
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