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Preparation method of electrochemical luminescence immunosensor for detecting beta-amyloid protein by isoluminol functionalization MOFs

A technology based on amyloid and isoluminol, applied in the field of preparation of electrochemiluminescent immunosensors, can solve problems such as surface plasmon resonance is easily interfered, cumbersome enzyme-linked immunoassay operation steps, poor controllability of fluorescence analysis, etc., to achieve Excellent catalytic properties and conductivity, high selectivity and high sensitivity detection, and the effect of improving stability

Inactive Publication Date: 2019-04-23
UNIV OF JINAN
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the operation steps of enzyme-linked immunoassay are cumbersome and time-consuming; fluorescence analysis has poor controllability and high toxicity; surface plasmon resonance is easily disturbed and has poor stability, etc.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Preparation method of an electrochemiluminescent immunosensor for detecting β-amyloid protein with isoluminol-functionalized MOFs

[0041] (1) Polish the glassy carbon electrode with a diameter of 4 mm to the mirror surface with alumina powder, and then wash it with ultrapure water;

[0042] (2) Drop-coat 6 µL of an aqueous solution of gold-functionalized ferric oxide-polypyrrole complex with a concentration of 5 mg / mL on the surface of the electrode, and store it at room temperature until dry;

[0043] (3) Continue to drip-coat 6 μL of β-amyloid capture antibody standard solution at a concentration of 1 μg / mL on the surface of the glassy carbon electrode, store in a refrigerator at 4 ºC until dry, and wash with ultrapure water;

[0044] (4) Continue to drip-coat 3 μL of bovine serum albumin with a mass fraction of 1% to block non-specific active sites, store in a refrigerator at 4 ºC until dry, and wash with ultrapure water;

[0045] (5) Continue to apply 6 ...

Embodiment 2

[0047] Example 2 Preparation method of an electrochemiluminescence immunosensor for detecting β-amyloid protein with isoluminol-functionalized MOFs

[0048] (1) Polish the glassy carbon electrode with a diameter of 4 mm to the mirror surface with alumina powder, and then wash it with ultrapure water;

[0049] (2) Drop-coat 6 µL of an aqueous solution of gold-functionalized ferric oxide-polypyrrole complex with a concentration of 10 mg / mL on the surface of the electrode, and store it at room temperature until dry;

[0050] (3) Continue to drip-coat 6 μL of β-amyloid capture antibody standard solution at a concentration of 1 μg / mL on the surface of the glassy carbon electrode, store in a refrigerator at 4 ºC until dry, and wash with ultrapure water;

[0051] (4) Continue to drip-coat 3 μL of bovine serum albumin with a mass fraction of 1% to block non-specific active sites, store in a refrigerator at 4 ºC until dry, and wash with ultrapure water;

[0052] (5) Continue to apply ...

Embodiment 3

[0054] Example 3 Preparation of complexes of isoluminol-functionalized cobalt-based MOFs, gold nanoparticles and detection antibodies

[0055] (1) Preparation of isoluminol-functionalized cobalt-based MOFs

[0056] Add 0.1 mmol of isoluminol and 0.4 mmol of 2-aminoterephthalic acid into 15 mL of N,N-dimethylformamide, stir well, then add 1 mL of glutaraldehyde with a mass fraction of 1% dropwise, Stir magnetically for 48 hours in the dark, then add 0.4 mmol cobalt nitrate, react in an oil bath at 100 ºC for 4 hours, centrifuge to obtain a precipitate, wash with ethanol and ultrapure water three times, place in a vacuum oven, and dry at 60 ºC Finally, isoluminol-functionalized cobalt-based MOFs were obtained;

[0057] (2) Preparation of composites of isoluminol-functionalized cobalt-based MOFs and gold nanoparticles

[0058] Add 0.5 mL of diethylene glycol diacrylate phthalate into 3 mL of ultrapure water, and disperse evenly by ultrasonication for 5 minutes, then add 5 mg of...

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Abstract

The invention relates to a preparation method of an electrochemical luminescence immunosensor for detecting beta-amyloid protein by isoluminol functionalization MOFs, and belongs to the field of an electrochemical luminescence sensor. A luminescence reagent functionalized MOFs is synthesized to be used as a signal marker; isoluminol is used as a luminescence reagent; hydrogen peroxide is used as aco-reaction agent; cobalt-based MOFs is used as a co-reaction accelerator; good catalyst performance of the cobalt-based MOFs on hydrogen peroxide is used; the luminescence reagent ECL signal amplification is realized; meanwhile, the loading capacity of the isoluminol is increased through the big specific surface area of the MOFs; the luminescence efficiency and the stability of the luminescencereagent are improved. The linear range on the beta-amyloid protein is 10 fg*mL<-1> to 100 fg*mL<-1>; the detection limit is 3 fg*mL<-1>.

Description

technical field [0001] The invention relates to a preparation method of an electrochemiluminescent immunosensor for detecting beta-amyloid protein with isoluminol functionalized MOFs. Specifically, a gold-functionalized ferric oxide-polypyrrole complex was used as a sensing platform, and a complex of isoluminol-functionalized cobalt-based MOFs and gold nanoparticles was used as a signal marker to prepare a detection method for detecting β-amyloid The invention relates to a signal-enhanced electrochemiluminescence sensor for a sample protein, belonging to the field of electrochemiluminescence sensors. Background technique [0002] With the acceleration of the global aging process, my country, as a country with a large population, has seen a surge in elderly patients suffering from Alzheimer's disease (AD). As a neurodegenerative disease that is difficult to prevent and cure, AD brings serious economic and living burdens to the elderly and their families. Abnormal aggregatio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N21/76G01N27/327G01N27/48
CPCG01N21/76G01N27/327G01N27/48
Inventor 魏琴王超魏东范大伟任祥王耀光吴丹马洪敏鞠熀先
Owner UNIV OF JINAN
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