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Antibody modified ZnSe nanocrystalline based kit for detection of hepatocellular carcinoma marker GPC3

An antibody modification and nanocrystal technology is applied in the field of kits for detecting liver cancer marker GPC3, which can solve the problems of poor stability, insufficient sensitivity and low ELISA detection sensitivity, and achieve the effects of stable signal, low detection limit and high sensitivity

Active Publication Date: 2014-10-08
FUZHOU HOSPITAL FOR INFECTIOUS DISEASE
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  • Description
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AI Technical Summary

Problems solved by technology

At present, the detection of serum GPC3 content is mainly carried out by ELISA. However, ELISA has many problems such as high price, poor stability, and insufficient sensitivity.
[0004] In recent years, various fluorescent nanomaterials have been widely used in biomedical detection, especially the combination of the specificity of immune response and the sensitivity of fluorescent nanomaterials, which can accurately, specifically, sensitively and quickly detect some trace or even ultra-trace substances , is expected to solve the problems of low sensitivity and poor stability in ELISA detection

Method used

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  • Antibody modified ZnSe nanocrystalline based kit for detection of hepatocellular carcinoma marker GPC3
  • Antibody modified ZnSe nanocrystalline based kit for detection of hepatocellular carcinoma marker GPC3
  • Antibody modified ZnSe nanocrystalline based kit for detection of hepatocellular carcinoma marker GPC3

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Kit composition:

[0026] ZnSe nanocrystals surface cross-linked modified GPC3 antibody, see below for the preparation method;

[0027] Zinc ion fluorescent probe solution: 10mM HEPES (pH7.3) + AgNO 3 500μM+FluoZin-33μM;

[0028] GPC3 antibody (adsorption antibody): mouse anti-human GPC3 monoclonal antibody (the recognition site of this antibody is the 121-220th amino acid of the reconstituted GPC3 protein, purchased from Sigma);

[0029] GPC3 protein standard product: human recombinant GPC3 protein (purchased from Life Company);

[0030] Coating buffer: carbonate buffer with pH=9.6;

[0031] Washing buffer: PBS solution with pH=7.4;

[0032] Blocking buffer: PBS solution with 1% BSA mass concentration (pH=7.4).

[0033] Preparation of ZnSe nanocrystals modified by surface cross-linking of GPC3 antibody:

[0034] (1) Weigh 1.10 grams of Se and 7.6 grams of Na respectively 2 SO 3 powder, add the two into 40mL deionized water (resistance of 18 megohms) and heat at...

Embodiment 2

[0037] Embodiment 2: the establishment of GPC3 detection standard curve

[0038] (1) GPC3 capture: add 100 μL, 10 μg / mL GPC3 antibody (absorbed antibody, dissolved in carbonic acid buffer solution) to a 96-well plate, incubate overnight at 4°C, block with blocking buffer after incubation, and then separate the gradient concentration The GPC3 protein standard solution was added to the microtiter plate and incubated at 37°C for 1 hour (at the same time, 100ng / mL OVA standard solution (purchased from Thermo) was added in parallel as an interference control, and the operation method was the same as that of the GPC3 protein standard solution). After multiple washes, add 100 μL of the ZnSe nanocrystal solution of the modified GPC3 detection antibody obtained in Example 1 (100 times diluted in PBS solution) to the above microtiter plate, and incubate at 37°C for 1 hour. After the incubation is completed, wash with PBS three times;

[0039] (2) Signal amplification and detection: Take...

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Abstract

The invention relates to an antibody modified ZnSe nanocrystalline based kit for detection of the hepatocellular carcinoma marker GPC3 (glypican-3). The kit mainly includes: a ZnSe nanocrystalline antibody with a GPC3 antibody crosslinking modified surface, a zinc ion fluorescent probe solution, a GPC3 antibody and a GPC3 protein standard substance. The kit provided by the invention combines a double-antibody sandwich immunoassay method and cation exchange reaction of ZnSe nanocrystalline, realizes amplification of the GPC3 detection signal in immunoreaction, has the advantages of stable signal, high sensitivity, low detection limit and the like in detection of hepatocellular carcinoma marker GPC3, and can be used for early diagnosis of hepatocellular carcinoma patients.

Description

(1) Technical field [0001] The invention relates to a kit for detecting liver cancer marker GPC3 based on antibody-modified ZnSe nanocrystals. (2) Background technology [0002] Hepatocellular carcinoma (HCC) is one of the most common malignant tumors clinically, and its incidence is increasing year by year. According to statistics, there are about 600,000 new patients with liver cancer every year in the world, and its fatality rate reaches the third place, only slightly behind lung cancer and gastric cancer. The number of new cases of hepatocellular carcinoma in my country accounts for about 55% of the world each year; the core of liver cancer prevention and treatment is early detection, early diagnosis and early treatment. Among them, early diagnosis is the most critical, that is, it can be diagnosed before the clinical symptoms of the patient or before the cancer infiltrates, so as to get timely treatment, so as to improve the cure rate and reduce the mortality rate. Ph...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N21/64
CPCG01N33/54373G01N33/57488
Inventor 刘景丰刘小龙韦学勇黄爱民王庆堂曾永毅
Owner FUZHOU HOSPITAL FOR INFECTIOUS DISEASE
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