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Tumor detection sample, preparation method therefor and application thereof

A technology for detecting substances and tumors, applied in the field of molecular biology, can solve the problems of low light sensitivity, easy quenching of fluorescent reporter molecules, etc., and achieve the effects of improving fluorescence intensity, shortening detection time, and facilitating screening.

Inactive Publication Date: 2019-03-29
ARMY MEDICAL UNIV
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Problems solved by technology

[0005] The present invention intends to provide a tumor detection substance and its preparation method and application to solve the problems of easy quenching of fluorescent reporter molecules and low photosensitivity in the prior art

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  • Tumor detection sample, preparation method therefor and application thereof

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Embodiment Construction

[0026] The following is further described in detail through specific implementation methods:

[0027] 3 kinds of blood group antibodies A antibody, B antibody and Rh(D) antibody used in the present invention come from abcam company; the secondary antibody used is phytohemagglutinin from Sigma company; superparamagnetic nano-microspheres come from NEB company; different types The carboxyl microspheres were purchased from Shanghai Toujing Technology Co., Ltd.; 1-ethyl-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC), N-hydroxysulfosuccinyl All imines (NHS) were purchased from Shanghai Sangon Biological Co., Ltd., and commercialized microspheres No. 9 were selected.

[0028] Buffer preparation:

[0029] Activation buffer: 100mM NaH2PO4, pH 6.3;

[0030] Coupling buffer: 50mM HEPES, pH 7.4;

[0031] Phosphate buffer solution (PBS): 10mM NaH2PO4, 150mM NaCl, pH 7.4;

[0032] 2-Morpholineethanesulfonic acid buffer (MES): 0.01M, pH 6.0;

[0033] 2-Morpholineethane...

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Abstract

The invention relates to the field of molecular biology, and particularly discloses a tumor detection sample, a preparation method therefor and an application thereof. The detection sample is a "magnetic bead-secondary antibody-antigen-capture antibody-microsphere" penta-complex. The preparation method of the tumor detection sample comprises: preparing a "secondary antibody-magnetic bead" complex;coupling the secondary antibody with superparamagnetic nanospheres; corresponding the secondary antibody to the antigen and the capture antibody respectively; and binding with the capture antibody ondifferent epitopes of the antigen respectively. The tumor detection sample of the invention is mainly applied to quantitatively detect a tumor marker and at the same time, quantitatively detect different antigens, and solves problems that a fluorescent reporter molecule in the prior art is easy to quench, photosensitivity is low, there is no accurate basis for a quantitative analysis, and detection flux has a certain limitation.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a tumor detection substance and its preparation method and application. Background technique [0002] With the continuous development of various new technologies in the field of biotechnology, high-throughput, accurate and rapid detection methods have become important research methods in the fields of bioscience and medicine. Most of the traditional high-throughput biomolecular detection technologies are based on biochip technology. At present, biochip technologies commonly used in the detection of biomacromolecules are gene chips and protein chips. However, due to the high cost of biochip preparation, long cycle time, and difficulty in popularization, the application of biochip technology has limitations. [0003] In recent years, the xMAP (FlexibleMulti-Analyte Profiling) technology developed by Luminex Corporation of the United States, following the gene chip and protein chip...

Claims

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Application Information

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IPC IPC(8): G01N33/574G01N33/543
CPCG01N33/54326G01N33/57484
Inventor 刘文斌韩飞陈洪强陈卿徐婷敖琳刘晋祎曹佳
Owner ARMY MEDICAL UNIV
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