A photoelectrochemical immunosensor for detecting ca125 and its preparation method and application

A CA125, immune sensor technology, applied in the field of biomedical detection, can solve the problems of high cost, time-consuming, low sensitivity, etc.

Active Publication Date: 2021-03-19
SOUTHEAST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

CA125 is one of the biomarkers used to diagnose ovarian cancer. Currently, methods for detecting this marker include radioimmunoassay, enzyme-linked immunosorbent assay, electrochemiluminescence, and chemiluminescence. Costly, time-consuming, and less sensitive

Method used

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  • A photoelectrochemical immunosensor for detecting ca125 and its preparation method and application
  • A photoelectrochemical immunosensor for detecting ca125 and its preparation method and application
  • A photoelectrochemical immunosensor for detecting ca125 and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Embodiment 1, quenching agent SiO 2 Preparation of @PDA complex

[0044] (1) SiO 2 Synthesis of nanoparticles: first synthesize SiO with a particle size of about 50nm according to the stober method 2 Nanoparticles, the specific method is: 80mL ethanol, 3.6mL ammonia water, 4.85mL ultrapure water in a single-necked flask, when the temperature rises to 55°C, add a mixture of 3.1mLTEOS and 8mL ethanol, stir for 5h, and obtain a slightly milky white gel solution , washed by centrifugation, and stored in freeze-dry.

[0045] (2) SiO 2 Synthesis of @PDA nanocomposite: then weighed 35mg SiO 2 , 60mg of dopamine hydrochloride and 6mg of Tris were dissolved in 25mL of ultrapure water and stirred for 72h to obtain SiO 2 @PDA nanocomposites.

[0046] (3) Ab 2 -SiO 2 Synthesis of @PDA nanocomplex: 60μL, 100ug / ml Ab 2 (in 0.02M PH=7.4PBS) was added to 400μL 0.1mg / mLSiO 2 Incubate at 4°C for 12h in @PDA (with gentle stirring).

[0047] Then add 1% BSA (dissolved in 0.02M p...

Embodiment 2

[0048] Embodiment 2, the photoelectric activity detection of quencher

[0049] (1) Instrument: Shanghai Chenhua electrochemical workstation (chi660e software), xenon lamp light source, multimeter

[0050] (2) Materials and reagents

[0051] ITO: indium tin oxide semiconductor electrode, the size is 0.4cm×0.4cm

[0052] Electrolyte: 50mM Ascorbic Acid in Phosphate Buffer

[0053] Reagent: Ab 2 , Ab 2 -PDA, Ab 2 -SiO 2 @PDA

[0054] (3) Method: Electrode treatment: place the ITO electrode in acetone, ethanol, and ultrapure water for 20 minutes, blow dry with nitrogen, test the front and back with a multimeter and mark it for later use.

[0055] Dropping sample: Add 5μM CdTe / CS, 1% glutaraldehyde, 10μg / mLAb1, 1% BSA, 10U / mLAg each 10μL dropwise on the electrode surface, then add Ab respectively 2 , Ab 2 -PDA, Ab 2 -SiO 2 @PDA, detection of electrode photocurrent intensity.

[0056] Test: The xenon lamp light source system provides light, and uses the three-electrode s...

Embodiment 3

[0060] Embodiment 3, the preparation method of photoelectric immunosensor

[0061] (1) The signal layer is fixed

[0062] A mixed solution (volume ratio 10:1) of CdTe (5 μM) and CS (0.05 wt %, solvent acetic acid) was added dropwise on the surface of the base electrode ITO by electrostatic adsorption, and placed at room temperature in a dry place for 12 hours to form a film. Dry at 80°C for 2 hours to further strengthen the film, then wash with deionized water and dry to obtain a CdTe / CS / ITO substrate electrode.

[0063] (2) Anchor recognition molecules

[0064] Glutaraldehyde was used as a cross-linking agent to connect Ab 1 with the base electrode. That is, dropwise add a mass fraction of 1% glutaraldehyde solution (the solvent is ultrapure water) to the surface of the base electrode obtained in step (1), leave it at room temperature for 1 hour, and then wash it with deionized water. After cleaning, the recognition molecule 10 μg / ml Ab 1 (The solvent is 0.02M pH=7.4PBS) ...

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Abstract

The invention discloses a photoelectrochemical immunosensor for detecting CA125, a preparation method and application thereof; on the basis of the principle that PDA can quench CdTe fluorescence on anelectrode substrate, by using of a SiO2@PDA nanocomposite as a probe molecule formed by self-polymerization deposition of dopamine DA and coating with SiO2 and using of CdTe as a photoelectrically-active substance, the photoelectrochemical immunosensor is constructed to detect tumor markers. According to the method, the photoelectrochemical immunosensor is constructed by use of the SiO2@PDA nanocomposite, and is used to detect the tumor markers, compared with an enzyme linked immunosorbent assay, a traditional optical method and the like, the photoelectrochemical immunosensor has the advantages of simple operation, low technical requirement and low price and easy miniaturization, and by use of fluorescence quencher SiO2@PDA, loading of biomolecules can be increased, and a signal can be amplified.

Description

technical field [0001] The invention belongs to the field of biomedical detection, and in particular relates to a photoelectrochemical immunosensor for detecting CA125, a preparation method and application thereof. Background technique [0002] Ovarian cancer is one of the most fatal gynecological malignancies in women, and its mortality rate is much higher than any other cancer of the female reproductive system. CA125 is one of the biomarkers used to diagnose ovarian cancer. Currently, the methods used to detect this marker include radioimmunoassay, enzyme-linked immunosorbent assay, electrochemiluminescence, and chemiluminescence. The cost is high, time-consuming, and the sensitivity is low. The photoelectrochemical immunoassay is a combination of high-sensitivity photoelectrochemical analysis and high-specificity immune reaction. In photoelectrochemical detection, light is used as an excitation source to excite photoelectrically active substances, and the electrical sign...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/574G01N33/94
CPCG01N33/57449G01N33/57484G01N33/9413G01N2800/36
Inventor 沈艳飞薛怀佳潘登张袁健
Owner SOUTHEAST UNIV
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