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Method for detecting serum biomarker of liver cancer patient

A technology for serum and antibody detection, applied in biological testing, measuring devices, material testing products, etc., can solve the problems of complex enzyme chain immunodetection technology, high price, insufficient sensitivity, etc., to achieve specific recognition and response, inhibition Release, excellent performance effect

Active Publication Date: 2020-12-18
QINGDAO UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The remarkable advantage of the present invention is that it overcomes the difficulties of traditional methods relying on peroxidase to generate signals and signal amplification, improves the detection sensitivity to the greatest extent, simplifies the detection process, reduces the detection cost, broadens the application field, and solves the problem of enzyme Chain immunoassay technology is complicated, the price is high, and the sensitivity is insufficient. It has potential application value in the fields of early diagnosis and treatment of tumors; the second purpose of the present invention is to propose a new type of AFP based on PDA and hollow gold nanoparticles. The preparation method of the signal-controlled release nanomaterial of enzyme-free immunodetection with its primary antibody and secondary antibody; the third purpose of the present invention: to provide a new type of non-enzymatic drug based on AFP and its primary antibody and secondary antibody using PDA and hollow gold nanometers. Method for detecting AFP using signal-controlled release nanomaterials for enzyme immunoassay

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  • Method for detecting serum biomarker of liver cancer patient

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Effect test

Embodiment 1

[0025] A preparation method for the novel novel method of utilizing PDA and hollow gold nanoparticles to construct a signal-controlled release nanomaterial based on enzyme-free immunodetection of AFP and its primary antibody and secondary antibody, comprising the following steps:

[0026] (1) Add 5 μL to 1 mL hollow gold nano-gold suspension with a concentration of 1.0×10 -5 mol / L RhB solution, 37°C shaker reaction for 12h;

[0027] (2) Centrifuge, remove the supernatant, and dissolve in 1mL of 10mM PBS buffer;

[0028] (3) Add the solution prepared in the previous step to 1 mL of 0.05 mg / mL dopamine Tris-HCl solution, and react at room temperature for 6 h;

[0029] (4) Centrifuge, add 100 μL of secondary antibody suspension with a concentration of 1 mg / mL, incubate at room temperature for 30 min, and place in a refrigerator at 4 °C overnight;

[0030] (5) The precipitate was collected by centrifugation and washed with PBS solution to obtain signal-controlled release nanomat...

Embodiment 2

[0033] A method for detecting AFP in a human serum sample using PDA and hollow gold nanoparticles to construct a signal-controlled release nanomaterial based on AFP and its primary antibody and secondary antibody without enzyme immunodetection, comprising the following steps:

[0034] (1) Incubate the well plate coated with the primary antibody with blocking buffer for 2 hours at room temperature, and wash the well plate;

[0035] (2) Add 50 μL of the dilution solution of the human serum sample obtained from the hospital, and incubate at 37°C for 1 hour;

[0036] (3) Wash the well plate, add 300 μL of the signal-controlled release nanomaterial solution prepared in Example 1, and incubate at 37° C. for 1 h;

[0037] (4) Wash the well plate, adjust the pH to 2.0 with 0.1M HCl, and after standing for 4 hours, take the supernatant for fluorescence detection, the excitation wavelength is 553nm, and the slit width is 5.0nm.

[0038] Among them, the test samples were obtained from 3...

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Abstract

The invention relates to a method for detecting a serum biomarker of a liver cancer patient. Alpha fetoprotein is closely related to generation and development of various tumors, is mainly used as a serum marker of liver cancer clinically, and is used for diagnosis and curative effect monitoring of primary liver cancer. Therefore, the high-sensitivity detection of the alpha fetoprotein is of greatsignificance to the early diagnosis and treatment of liver cancer patients. The invention provides a method for detecting alpha fetoprotein in human serum by using a novel signal controlled-release nano material for enzyme-free immunoassay based on alpha fetoprotein and primary and secondary antibodies thereof. The method not only overcomes the difficulties that the traditional method depends onperoxidase to generate signals and amplify the signals, but also remarkably improves the detection sensitivity, simplifies the detection process, reduces the cost, widens the application field, and solves the problems of complex enzyme-linked immunosorbent assay technology, high price, insufficient sensitivity and the like; the method has a wide application prospect in the fields of early diagnosis and treatment of tumors and the like, and a novel method with high sensitivity and high specificity is provided for enzyme-free immunodetection.

Description

technical field [0001] The invention relates to the detection of serum biomarkers of liver cancer patients, in particular to a method for detecting serum biomarkers of tumor patients AFP, which belongs to the fields of photoelectrochemical biosensing analysis and biomedical clinical diagnosis. Background technique [0002] Alpha-fetoprotein (AFP) is a glycoprotein and an important tumor marker. Under normal circumstances, this protein mainly comes from the liver cells of the embryo. About two weeks after the fetus is born, alpha-fetoprotein disappears from the blood. Therefore, the level of alpha-fetoprotein in normal human serum is lower than 20ng / mL, but it rises significantly in the serum of patients with liver cancer. High, become a specific marker of liver cancer. In addition, alpha-fetoprotein is also closely related to the occurrence and development of various tumors, such as testicular cancer, ovarian tumor, malignant teratoma, pancreatic cancer, gastric cancer, int...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/58G01N33/543G01N33/533B82Y15/00
CPCG01N33/57438G01N33/57476G01N33/582G01N33/54306G01N33/533B82Y15/00
Inventor 王卫李欣唐凯
Owner QINGDAO UNIV OF SCI & TECH
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