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Preparation method of quantum dot based prostate specific antigen immuno-chromatographic test strip

A prostate-specific, chromatographic test paper technology, applied in the field of medical diagnosis, can solve the problems of inability to complete quantitative detection, the color of the detection strip cannot be identified with the naked eye, etc., and achieve the effect of improving the ability to resist non-specific adsorption and low price.

Inactive Publication Date: 2015-11-11
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The commercialized immunochromatographic test strips mostly use colloidal gold as a marker. Although the test results are visible to the naked eye, quantitative detection cannot be completed, and the test strips of samples with low antigen concentration are too light to be recognized by the naked eye.

Method used

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  • Preparation method of quantum dot based prostate specific antigen immuno-chromatographic test strip

Examples

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Embodiment 1

[0046] 1. Prostate-specific antibody conjugated to quantum dots

[0047] (1) Water-soluble quantum dots, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) aqueous solution are mixed in phosphate buffer according to the mass ratio of raw materials at a ratio of 1:4000 (PBS buffer solution), rotate at a low speed on a vortex to make it evenly mixed, then place the above mixed solution on a rotating mixing rack, and react at room temperature for 30 minutes;

[0048] (2) After the reaction, purify by centrifugation, and wash at least three times with PBS buffer to remove unreacted 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC ), to obtain activated water-soluble quantum dots;

[0049] (3) Mix the activated water-soluble quantum dots and α-prostate cancer antigen-specific monoclonal antibody (ab140337) in phosphate buffer (PBS buffer) at a molar ratio of 1:20, and rotate on a vortex at a low speed Make it evenly mixed, place the above mixed solu...

Embodiment 2

[0065] 1. Prostate-specific antibody conjugated to quantum dots

[0066] (1) Water-soluble quantum dots, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) aqueous solution are mixed in phosphate buffer according to the ratio of raw material mass ratio of 1:7000 (PBS buffer solution), rotate at a low speed on a vortex to make it evenly mixed, then place the above mixed solution on a rotating mixing rack, and react at room temperature for 30 minutes;

[0067] (2) After the reaction, purify by centrifugation, and wash at least three times with PBS buffer to remove unreacted 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC ), to obtain activated water-soluble quantum dots;

[0068] (3) Mix the activated water-soluble quantum dots and α-prostate cancer antigen-specific monoclonal antibody (ab403) in phosphate buffer (PBS buffer) at a molar ratio of 1:30, and rotate at a low speed on a vortex instrument Make it evenly mixed, place the above mixed so...

Embodiment 3

[0083] 1. Prostate-specific antibody conjugated to quantum dots

[0084] (1) Water-soluble quantum dots, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) aqueous solution are mixed in phosphate buffer according to the ratio of raw material mass ratio of 1:10000 (PBS buffer solution), rotate at a low speed on a vortex to make it evenly mixed, then place the above mixed solution on a rotating mixing rack, and react at room temperature for 30 minutes;

[0085] (2) After the reaction, purify by centrifugation, and wash at least three times with PBS buffer to remove unreacted 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC ), to obtain activated water-soluble quantum dots;

[0086] (3) Mix the activated water-soluble quantum dots and α-prostate cancer antigen-specific monoclonal antibody (ab63590) in phosphate buffer (PBS buffer) at a molar ratio of 1:40, and rotate at a low speed on a vortex instrument Mix it evenly, place the above mixed soluti...

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Abstract

The invention relates to a preparation method of a quantum dot based prostate specific antigen immuno-chromatographic test strip. The test strip is composed of a sample pad, a bonder release pad, a chromatography membrane, and a water absorbing pad, wherein the sample pad and bonder release pad are prepared from preprocessed polyester fibers, and the chromatography membrane and water absorbing pad are made of a preprocessed cellulose nitrate membrane. A sample is dropwise added on the sample pad. The surface of the bonder release pad is painted with quantum dot marked alpha-prostate cancer antigen specific monoclonal antibody. Beta-prostate cancer antigen specific monoclonal antibody and goat-anti-mouse second antibody are coated in the chromatography membrane to form a T line (detection line) and a C line (quality control line). The water absorbing pad drives the liquid to be detected to flow. Compared to the conventional products and technologies, the provided preparation method has the advantages that the whole preparation process is simple, and the preparation method is suitable for industrial production. According to the fluorescent characteristics of quantum dots, the prostate specific antigen can be detected quantitatively and qualitatively, and the specificity is high. The operation is very convenient during the whole process, and the cost is low, so the provided test strip can be applied to tumor screening of high risk groups in communities. A novel method for detecting tumor is established.

Description

technical field [0001] The invention relates to the technical field of medical diagnosis, and more specifically relates to a preparation method of a novel quantum dot-based immunochromatographic test strip for detecting prostate specific antigen. Background technique [0002] Prostate cancer (PCa) is a common malignant tumor of the genitourinary system in elderly men. In developed countries in Europe and the United States, prostate cancer ranks second in male malignant tumors, and its case fatality rate is second only to lung cancer. In recent years, the incidence of prostate cancer in my country has been on the rise, ranking third among urological tumors, and the age of onset is also getting younger. Clinically, it has been regarded as one of the common malignant tumors in elderly men. However, if patients can be found in the early stage of the disease, the cure rate can reach more than 80%. Therefore, the early diagnosis of prostate cancer has become a research focus in r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574
CPCG01N33/57434
Inventor 常津姚颖异宫晓群刘俊庆武玉东张健
Owner TIANJIN UNIV
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