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Method for preparing optical sensitive film for fixing human brain natriuretic peptide (BNP)

A technology of brain natriuretic peptide and sensitive membrane, applied in the field of clinical medical diagnosis, can solve the problems of increased detection cost and achieve the effect of rapid detection

Inactive Publication Date: 2010-12-22
INST OF ELECTRONICS CHINESE ACAD OF SCI
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  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the detection of BNP in my country mainly relies on imported instruments and reagents, and the resulting detection costs have also increased significantly

Method used

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  • Method for preparing optical sensitive film for fixing human brain natriuretic peptide (BNP)
  • Method for preparing optical sensitive film for fixing human brain natriuretic peptide (BNP)
  • Method for preparing optical sensitive film for fixing human brain natriuretic peptide (BNP)

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preparation example Construction

[0029] A kind of preparation method of the optically sensitive film of fixing human brain natriuretic peptide of the present invention, comprises the following steps:

[0030] 1. Prepare the conjugate of fluorescent quantum dots and BNP detection antibody, and the detection antibody can recognize the inner ring structure of BNP molecule.

[0031] 2. Select the carrier, the carrier can be one of biological slides or ordinary glass or PC plate or organic polymer film.

[0032] Such as figure 2 Shown:

[0033] 3. Surface treatment: use silane as a surfactant to aminate the surface of the glass sheet, the amino group can be combined with the bifunctional reagent glutaraldehyde, and the glutaraldehyde is coupled with the capture antibody to achieve the immobilization of the BNP capture antibody.

[0034] 4. Immobilize the antibody: Add the BNP capture antibody that can recognize the C-terminal region of the BNP molecule dropwise on the surface of the aldylated carrier, place it ...

Embodiment

[0040] 1) Preparation of fluorescent quantum dots and BNP antibody conjugates

[0041] Pipette 20 μl (2nmol / L) of avidin-labeled cadmium selenide (CdSe) fluorescent quantum dots into a 2ml centrifuge tube, add 100μl Tris-HCl buffer (containing 1mmol / LEDTA, 1mol / L NaCl, pH=7.4), 10 μl of biotin-labeled BNP antibody was dissolved in 80 μl of Tris-HCl buffer solution, and the antibody concentration at this time was 0.15 mg / mL. The above biotin-labeled antibody solution was added to the avidin-modified fluorescent quantum dot solution, and reacted in a shaker at 37° C. and 150 rpm / min for 30 minutes. Add 1 ml of PBST buffer solution (containing 0.05% Tween-20 and 0.1% BSA), and centrifuge to remove the BNP antibody solution not involved in the reaction. Add PBS buffer to prepare a solution with a concentration of 2.5 mg / mL fluorescent quantum dot-labeled BNP antibody.

[0042] 2) BNP capture antibody immobilized on aldehyde-modified glass substrate

[0043] Such as figure 1 As s...

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Abstract

The invention discloses a method for preparing an optical sensitive film for fixing the human brain natriuretic peptide (BNP) and relates to a clinical medical diagnosis technology. The optical sensitive film can be used for detecting the BNP. The method comprises the following steps of: selecting a vector; carrying out surface treatment; fixing a captured antibody; sealing and capturing antigen; adding a detection antibody marked by a luminescent quantum dot to form a double-antibody sandwich compound, and the like, wherein the surface of the vector is treated through an aldehyde reagent, and the NBP captured antibody molecule is fixed on the surface of the vector in a mode of a covalent bond through a difunctional cross-linking agent. Because the sensitive film prepared by the method comprises the vector of which the surface is treated, the BNP captured antibody, the BNP antigen and the BNP detection antibody marked with the luminescent quantum dot, the sensitive film can be used for quantitatively detecting the content of the human BNP. The method has the advantages of simple operation and convenient finished product use and is suitable for the quantitative detection to the content of the human BNP clinically.

Description

technical field [0001] The invention relates to the technical field of clinical medical diagnosis, and relates to a method for preparing an optically sensitive film for immobilizing human brain natriuretic peptide. The sensitive film can accurately, rapidly and quantitatively detect the content of human brain natriuretic peptide. Background technique [0002] With the improvement of people's living standards, accelerated pace of life, and increased psychological pressure, the incidence of heart failure has increased year by year, and it has become a common disease in modern society. Experts pointed out that in various cardiovascular and cerebrovascular accidents, we should be particularly alert to heart failure, because the damage to the heart caused by heart failure is irreversible. To prevent heart failure, the best way is early detection and early treatment. But from the current point of view, because heart failure has no obvious clinical symptoms, 50% of heart failure p...

Claims

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Application Information

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IPC IPC(8): G01N33/533G01N33/553G01N33/577G01N21/64
Inventor 刘儒平蔡新霞刘军涛王蜜霞刘春秀罗金平
Owner INST OF ELECTRONICS CHINESE ACAD OF SCI
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