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Preparation method of AFP (alpha fetoprotein) immunochromatographic test strip based on QDs (quantum dots)

An immunochromatographic test strip and alpha-fetoprotein technology, which is applied in the field of medical diagnosis, can solve the problems of limiting the application of serum tumor markers, the color of the detection band cannot be identified with the naked eye, etc., and achieves qualitative and quantitative detection, low price, and sensitivity. high effect

Inactive Publication Date: 2015-10-21
TIANJIN UNIV
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Problems solved by technology

However, colloidal gold is used as a marker, and the detection bands of samples with low antigen concentration a

Method used

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  • Preparation method of AFP (alpha fetoprotein) immunochromatographic test strip based on QDs (quantum dots)
  • Preparation method of AFP (alpha fetoprotein) immunochromatographic test strip based on QDs (quantum dots)
  • Preparation method of AFP (alpha fetoprotein) immunochromatographic test strip based on QDs (quantum dots)

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Example Embodiment

[0046] The preparation method is characterized in that the quantum dots are water-soluble quantum dots obtained by modifying CdZnSe quantum dots with poly(tert-butyl acrylate-ethyl acrylate-methacrylic acid) triblock copolymer. (Such as figure 1 Shown)

[0047] The preparation method is characterized in that the mass fraction ratio of the quantum dots to 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) is 1:4000-10000.

[0048] The preparation method is characterized in that the molar ratio of the raw materials: quantum dots: AFP monoclonal antibody Ab1=1:10-100.

[0049] The preparation method is characterized in that the hydrophilic polymer in the treatment liquid formulation is one of PEG-4000, PVP-10000, and PVP-40000.

[0050] The preparation method is characterized in that the surfactant in the formula of the treatment liquid is Triton X-100 and Tween-20.

[0051] The preparation method is characterized in that the concentration of BSA in the treatment solution f...

Example Embodiment

[0052] Implementation case 1:

[0053] 1. Quantum dot coupled alpha-fetoprotein antibody (QDsAb1)

[0054] (1) Mix the water-soluble quantum dots and the aqueous solution of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) in the phosphate buffer at a ratio of 1:4000 to the mass ratio of the raw materials (PBS buffer), activate the carboxyl groups of the quantum dots on a rotating mixing rack for 30 minutes, and centrifuge to obtain activated quantum dots.

[0055] (2) Mix the activated quantum dots and alpha-fetoprotein antibody Ab1 in PBS buffer at a molar ratio of 1:20, place the mixed solution on a rotating mixing rack, and react for 2 hours at room temperature;

[0056] (3) After the reaction, use centrifugal separation to purify, wash three times with PBS buffer to remove free antibodies and EDC to obtain fluorescent probe QDsAb1, and finally disperse the product in PBS containing 1% bovine serum albumin (BSA) In the buffer, let stand overnight at 4°C.

[0057] ...

Example Embodiment

[0069] Implementation case 2:

[0070] 1. Quantum dot coupled alpha-fetoprotein antibody (QDsAb1)

[0071] (1) Mix the water-soluble quantum dots and the aqueous solution of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) in the phosphate buffer at a ratio of 1:6000 to the mass ratio of the raw materials (PBS buffer), activate the carboxyl groups of the quantum dots on a rotating mixing rack for 30 minutes, and centrifuge to obtain activated quantum dots.

[0072] (2) Mix the activated quantum dots and the alpha-fetoprotein antibody Ab1 in the PBS buffer at a molar ratio of 1:60, place the mixed solution on a rotating mixing rack, and react for 2.5 hours at room temperature;

[0073] (3) After the reaction, use centrifugal separation to purify, wash three times with PBS buffer to remove free antibodies and EDC to obtain fluorescent probe QDsAb1, and finally disperse the product in PBS containing 2% bovine serum albumin (BSA) In the buffer, let stand overnight at 4°C...

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Abstract

The invention relates to a preparation method of an AFP (alpha fetoprotein) immunochromatographic test strip based on QDs (quantum dots). The preparation method comprises the following steps: QDs and an AFP antibody are coupled to prepare a fluorescent probe QDs@Ab1; the QDs@Ab1, AFP and another AFP antibody (Ab1') embedded in a test strip form a sandwich-like structure QDs@Ab1-AFP-Ab1' through the action between antibodies and antigens; the tumor marker AFP of primary hepatic carcinoma is qualitatively and half-quantitatively detected. Compared with the conventional product and technology, the test strip has the following advantages: the whole preparation process is simple and applicable to industrial production; the antigen concentration is in positive correlation with the fluorescence intensity of the probe, so that the strip can qualitatively and quantitatively detect AFP; the detecting process is low in cost, quite easy and convenient to operate and suitable for tumor screening in communities with high risk groups; a novel method for detecting tumor is established.

Description

technical field [0001] The invention relates to the technical field of medical diagnosis, in particular to a preparation method of a novel quantum dot-based alpha-fetoprotein immunochromatographic test strip. Background technique [0002] Primary liver cancer (Primary Hepatic Carcinoma PHC) is one of the most common malignant tumors clinically. At present, the incidence of liver cancer in the world is on the rise. According to the "Global Cancer Report 2014" published by the World Health Organization, the number of new cancer cases in China ranks first in the world, and the number of new cases and deaths of liver cancer ranks first in the world. At present, the incidence rate of liver cancer in my country is about 25.7 / 100,000, making it the third most deadly malignant tumor after gastric cancer and lung cancer. Therefore, the diagnosis and treatment of primary liver cancer are of great significance. [0003] Alpha-fetoprotein (α-fetoprotein or AFP) is a glycoprotein. Und...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/574G01N33/558G01N33/533G01N33/52
CPCG01N33/577G01N33/523G01N33/533G01N33/558G01N33/57438G01N2333/47
Inventor 常津张健宫晓群武玉东姚颖异
Owner TIANJIN UNIV
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