A preparation method of electrochemiluminescence sensor based on protein activity protection

Abstract

The invention relates to a preparation method and application of an electrochemiluminescence sensor based on protein activity protection, and belongs to the technical field of electrochemiluminescence detection. Developed and verified for the first time the high-efficiency electrochemiluminescence behavior that europium-doped gadolinium phosphate can achieve under low potential excitation. On the one hand, it solved the problem of fixing the luminescent material on the electrode, and on the other hand, it solved the problem of effective preservation of antigen-antibody activity. The detection of procalcitonin is realized according to the difference in intensity of electrochemiluminescent signal in response to different concentrations of procalcitonin. The accuracy and precision of this method are demonstrated by using F test and T test, and the test results are all less than the theoretical value, which shows that the method is accurate and reliable.

Description

technical field

[0001] The invention relates to the preparation and application of a sensor based on europium-doped gadolinium phosphate as a new low-potential electrochemiluminescent emitter and energy transfer with core-shell palladium-functionalized cuprous oxide nanocrystals, specifically using europium-doped phosphoric acid Gadolinium hollow spheres are used as luminescent materials, palladium-functionalized cuprous oxide is used as a quencher, and polypeptide chains are used to achieve directional immobilization of antibodies, which realizes excitation at a low potential of -1.15 V, effectively protects the activity of proteins, and improves immunity. Feasibility of substance detection. In summary, a quenching-type electrochemiluminescence sensor for detecting procalcitonin prepared above belongs to the technical field of electrochemiluminescence detection. Background technique

[0002] SIRS refers to systemic inflammatory response, that is, the body's response to vari...

Examples

Embodiment 1

[0037] The preparation of embodiment 1 europium-doped gadolinium phosphate

[0038] (1) Preparation of phenolic resin template spheres

[0039] Add 0.1806 g of 3-aminophenol and 93 µL of 25% ammonia water into 50.4 mL of deionized water, and stir at 30°C for 10 min to form a clear solution. After adding 123 µL of formaldehyde solution, the solution turned white after 30 s. After stirring continuously for another 4 h, the resulting mixture was transferred into a 100 mL Teflon-lined stainless steel autoclave and kept at 100 ºC for 24 h. Finally, the sample was centrifuged and purified by washing twice with deionized water and alcohol, and finally dried in air;

[0040] (2) Preparation of europium-doped gadolinium phosphate

[0041] First, the precursors were prepared: 3 g of urea, 1 mmol of gadolinium nitrate aqueous solution, 0.05 mmol of europium nitrate aqueous solution and 0.2 g of the phenolic resin balls synthesized above were added to 25 mL of deionized water, respecti...

Embodiment 2

[0042] Example 2 Preparation of palladium-functionalized cuprous oxide-labeled procalcitonin capture antibody incubation solution

[0043] (1) Preparation of palladium nanocrystals

[0044] First, 0.048 g of cetyltrimethylammonium chloride, 9.125 mL of deionized water, and 0.7 mL of 10 mM chloropalladium acid solution were injected into the vial, which was stored in a 35ºC water bath. Subsequently, 500 µL of 1 mM potassium bromide solution and 50 µL of 1 mM potassium iodide solution were added and mixed. After 10 min, inject 1.2 mL of 0.05 M ascorbic acid, keep the water bath for 30 min, and centrifuge at 7500 rpm for 10 min twice. Final product in 400 µL of deionized water;

[0045] (2) Preparation of palladium-functionalized cuprous oxide

[0046] Dissolve 0.087 g sodium lauryl sulfate in deionized water, add 0.07 mL of 0.1 M copper chloride solution and 0.08 mL of palladium nanocrystal solution prepared in the previous step, and inject 0.25 mL of 1.0 M sodium hydroxide s...

Embodiment 3

[0049] Example 3 Preparation of palladium-functionalized cuprous oxide-labeled procalcitonin capture antibody incubation solution

[0050] (1) Preparation of palladium nanocrystals

[0051] First, 0.0100 g of cetyltrimethylammonium chloride, 9.125 mL of deionized water, and 0.2 mL of 10 mM chloropalladium acid solution were injected into the vial, which was stored in a 35ºC water bath environment. Subsequently, 300 µL of 1 mM potassium bromide solution and 50 µL of 1 mM potassium iodide solution were added and mixed. After 10 min, 0.5 mL of 0.05 M ascorbic acid was injected. Keep the water bath for 30 min and centrifuge at 7500 rpm for 10 min twice. Final product in 400 µL of deionized water;

[0052] (2) Preparation of palladium-functionalized cuprous oxide

[0053] Dissolve 0.020 g sodium lauryl sulfate in deionized water, add 0.03 mL of 0.1 M copper chloride solution and 0.02 mL of palladium nanocrystal solution prepared in the previous step, and inject 0.25 mL of 1.0 M...

technical field

[0001] The invention relates to the preparation and application of a sensor based on europium-doped gadolinium phosphate as a new low-potential electrochemiluminescent emitter and energy transfer with core-shell palladium-functionalized cuprous oxide nanocrystals, specifically using europium-doped phosphoric acid Gadolinium hollow spheres are used as luminescent materials, palladium-functionalized cuprous oxide is used as a quencher, and polypeptide chains are used to achieve directional immobilization of antibodies, which realizes excitation at a low potential of -1.15 V, effectively protects the activity of proteins, and improves immunity. Feasibility of substance detection. In summary, a quenching-type electrochemiluminescence sensor for detecting procalcitonin prepared above belongs to the technical field of electrochemiluminescence detection. Background technique

[0002] SIRS refers to systemic inflammatory response, that is, the body's response to vari...

Examples

Embodiment 1

[0037] The preparation of embodiment 1 europium-doped gadolinium phosphate

[0038] (1) Preparation of phenolic resin template spheres

[0039] Add 0.1806 g of 3-aminophenol and 93 µL of 25% ammonia water into 50.4 mL of deionized water, and stir at 30°C for 10 min to form a clear solution. After adding 123 µL of formaldehyde solution, the solution turned white after 30 s. After stirring continuously for another 4 h, the resulting mixture was transferred into a 100 mL Teflon-lined stainless steel autoclave and kept at 100 ºC for 24 h. Finally, the sample was centrifuged and purified by washing twice with deionized water and alcohol, and finally dried in air;

[0040] (2) Preparation of europium-doped gadolinium phosphate

[0041] First, the precursors were prepared: 3 g of urea, 1 mmol of gadolinium nitrate aqueous solution, 0.05 mmol of europium nitrate aqueous solution and 0.2 g of the phenolic resin balls synthesized above were added to 25 mL of deionized water, respecti...

Embodiment 2

[0042] Example 2 Preparation of palladium-functionalized cuprous oxide-labeled procalcitonin capture antibody incubation solution

[0043] (1) Preparation of palladium nanocrystals

[0044] First, 0.048 g of cetyltrimethylammonium chloride, 9.125 mL of deionized water, and 0.7 mL of 10 mM chloropalladium acid solution were injected into the vial, which was stored in a 35ºC water bath. Subsequently, 500 µL of 1 mM potassium bromide solution and 50 µL of 1 mM potassium iodide solution were added and mixed. After 10 min, inject 1.2 mL of 0.05 M ascorbic acid, keep the water bath for 30 min, and centrifuge at 7500 rpm for 10 min twice. Final product in 400 µL of deionized water;

[0045] (2) Preparation of palladium-functionalized cuprous oxide

[0046] Dissolve 0.087 g sodium lauryl sulfate in deionized water, add 0.07 mL of 0.1 M copper chloride solution and 0.08 mL of palladium nanocrystal solution prepared in the previous step, and inject 0.25 mL of 1.0 M sodium hydroxide s...

Embodiment 3

[0049] Example 3 Preparation of palladium-functionalized cuprous oxide-labeled procalcitonin capture antibody incubation solution

[0050] (1) Preparation of palladium nanocrystals

[0051] First, 0.0100 g of cetyltrimethylammonium chloride, 9.125 mL of deionized water, and 0.2 mL of 10 mM chloropalladium acid solution were injected into the vial, which was stored in a 35ºC water bath environment. Subsequently, 300 µL of 1 mM potassium bromide solution and 50 µL of 1 mM potassium iodide solution were added and mixed. After 10 min, 0.5 mL of 0.05 M ascorbic acid was injected. Keep the water bath for 30 min and centrifuge at 7500 rpm for 10 min twice. Final product in 400 µL of deionized water;

[0052] (2) Preparation of palladium-functionalized cuprous oxide

[0053] Dissolve 0.020 g sodium lauryl sulfate in deionized water, add 0.03 mL of 0.1 M copper chloride solution and 0.02 mL of palladium nanocrystal solution prepared in the previous step, and inject 0.25 mL of 1.0 M...