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A monochromatic ECL immunoassay method based on green light radiation quantum dots

Active Publication Date: 2018-04-27
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Although the above-mentioned ECL immunoassay method using quantum dots as markers has high sensitivity and accuracy, most of the quantum dots used are near-infrared quantum dots with ECL radiation around 700nm, which still belong to the red light region, and the detection range is still beyond reach. Extend to green light area

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  • A monochromatic ECL immunoassay method based on green light radiation quantum dots
  • A monochromatic ECL immunoassay method based on green light radiation quantum dots
  • A monochromatic ECL immunoassay method based on green light radiation quantum dots

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Embodiment 1

[0070] A monochromatic ECL immunoassay method based on green light radiation quantum dots, comprising the following steps:

[0071] (1) Preparation of CdSe quantum dot-labeled secondary antibody

[0072] Add 20 μL of phosphate containing 100 mg / mL 1-ethyl-3,3-dimethylaminopropylcarbodiimide (EDC) to 500 μL of an aqueous solution of mercaptopropionic acid and sodium hexametaphosphate-coated CdSe quantum dots Buffer solution and 20 μL phosphate buffer solution containing 100 mg / mL N-hydroxysuccinimide, reacted at room temperature for 30 minutes; after centrifugation and purification at 12000 rpm, take the precipitate and add 500 μL phosphate buffer solution for ultrasonic dispersion, then add 50 μL 1.0 μg / mL mL of secondary antibody, shake at room temperature for 2 hours; then add 20 μL of 1% volume concentration of bovine serum albumin, incubate at room temperature for 1 hour, centrifuge and purify at 12000 rpm, take the precipitate and add 500 μL of phosphate buffer solution f...

Embodiment 2-14

[0089] A monochromatic ECL immunoassay method based on green light radiation quantum dots, the steps are the same as in Example 1, the difference is:

[0090] In step (2) c, the carcinoembryonic antigen is respectively used as alpha-fetoprotein antigen, carbohydrate antigen 125, carbohydrate antigen 15-3, carbohydrate antigen 72-4, carbohydrate antigen 19-9, prostate specific antigen, free prostate Specific antigen, AIDS antigen, hepatitis B surface antigen, hepatitis B e antigen, thyroglobulin, troponin T antigen or myoglobin antigen instead.

Embodiment 15

[0092] A monochromatic ECL immunoassay method based on green light radiation quantum dots, the steps are the same as in Example 1, except that the volume fraction of bovine serum albumin used is 2%.

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Abstract

The invention discloses a monochromatic ECL immunodetection method based on green light radiation quantum dots, which mainly includes (1) preparation of CdSe quantum dot-labeled secondary antibody (CdSe QDs-Ab2), (2) using CdSe quantum dots as markers The preparation of the single-color ECL immunosensor and (3) drawing the working curve, carry out three steps of single-color ECL immunodetection. The detection sensitivity of the method of the invention is high, the detection limit reaches 0.1fg / mL, and single-molecule detection of the antigen can be realized; the selectivity is high. The electrochemiluminescent immunosensor constructed in the present invention is based on the specific recognition and binding between the antigen and the antibody, so the interfering protein in the test solution cannot combine with the first antibody and the second antibody of the antigen. No distractions.

Description

technical field [0001] The present invention relates to an electrogenerated chemiluminescence (electrogenerated chemiluminescence, ECL) immunoassay method, in particular to an immunoassay using highly passivated CdSe quantum dots as markers on the surface that can produce monochromatic electrochemiluminescence radiation in the green light region. Detection method. Background technique [0002] At present, most ECL kits and clinical diagnostic techniques use Ru(bpy) 3 2+ as an ECL marker. Given Ru(bpy) 3 2+ The ECL radiation spectrum of the system is wide, and it is located in the red light region. The development of an efficient monochromatic ECL system in the non-red light region is of great value for the basic and applied research of ECL analysis. [0003] Quantum dots are a new type of ECL emitter, and their unique photoelectric properties provide the possibility for the development of ECL systems in non-red regions. Liang et al. developed a near-infrared ECL immuno...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/76G01N33/68
Inventor 邹桂征张新
Owner SHANDONG UNIV
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