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Antibody chip kit for detecting cell adhesion molecules

A technology of adhesion molecules and antibody chips, which is applied in biological testing, measuring devices, and analytical materials, etc., can solve the problems of low sensitivity, single detection index, cumbersome operation, etc., and achieve the effect of less sample consumption and high detection sensitivity

Active Publication Date: 2017-07-04
RAYBIOTECH INC GUANGZHOU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] The purpose of the present invention is to overcome the deficiencies of the prior art and provide an antibody chip kit for detecting cell adhesion molecules, which can simultaneously detect multiple clinically used cell adhesion factors and overcome the cumbersome operation of the prior art , single detection index, low sensitivity and other defects, it has the advantages of cheap, convenient, sensitive, accurate, high-throughput, less specimen consumption, and can be popularized and scaled in ordinary laboratories.

Method used

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  • Antibody chip kit for detecting cell adhesion molecules
  • Antibody chip kit for detecting cell adhesion molecules
  • Antibody chip kit for detecting cell adhesion molecules

Examples

Experimental program
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Effect test

Embodiment 1

[0025] Screening of antibody chip carriers: Conventional antibody chips mostly use nitrocellulose membrane as a carrier. Since the nitrocellulose membrane has a multi-layer structure, it is difficult to wash the chip, resulting in a high background of the chip and large fluctuations in the results. At the same time, the nitrocellulose membrane is fragile, not easy to operate on a large scale, and it is not widely used for large-scale clinical samples. Traditional glass slides are cheap and have low background, which has brought breakthroughs in the widespread use of diagnostic and research fields. We screened the glass slides on the market that were not treated with the active method, and the spotting effect of the glass slides, whether it was aldehydes or aminations, was unstable. After a lot of screening, it is found that the temperature and the active reagent components on the surface of the slide are the key to the effect of spotting on the slide.

[0026] The following 5...

Embodiment 2

[0032] Preparation of an antibody chip kit for quantitative detection of multiple cell adhesion factors at the same time: In order to detect whether there are corresponding receptors in the sample, prepare slides immobilized with specific antibodies corresponding to 17 kinds of cell adhesion factors, 17 kinds of cells The specific antibody corresponding to the adhesion factor is a common commercial product.

[0033] Preparation and storage of the antibody chip: 100-1000 pl of PBS buffer solution containing specific antibodies (containing 0.01-10 g / 100 ml bovine albumin) was spotted on a glass slide with an automatic spotting device. The specific chip array uses biotin-labeled bovine IgG as a positive control. There are four replicate spots in each array for 17 antibodies and two different concentrations of positive controls. In this embodiment, the chip array adopts such as figure 1 The arrangement shown, but in fact, in other embodiments, the chip array can also be combined...

Embodiment 3

[0038] The experimental steps for quantitative detection of adhesion factors with the kit of the present invention are as follows:

[0039] 1. Complete drying of the slide chip: Take the slide chip out of the box, and after equilibrating at room temperature for 20-30 minutes, open the packaging bag, uncover the sealing strip, and then put the chip in a vacuum desiccator or dry at room temperature for 1-20 minutes. 2 hours.

[0040] 2. According to Table 2, carry out gradient dilution of cell adhesion factors

[0041] 2.1. Add 500 μl of sample diluent to the small tube of cell adhesion factor standard mixture, and redissolve the standard. Before opening the vial, give it a quick centrifuge and gently pipet up and down to dissolve the powder. Label this vial as Std1.

[0042] 2.2. Mark 6 clean centrifuge tubes as Std2, Std3 to Std7 respectively, and add 200 μl of sample diluent to each small tube.

[0043] 2.3. Take 100 μl of Std1 and add it to Std2 and mix gently, then take ...

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Abstract

The present invention relates to the technical field of biological detection, particularly to an antibody chip kit for detecting cell adhesion molecules. According to the antibody chip kit, a glass slide treated with a hydrophilic reagent containing alkyl glycoside as a solid-phase carrier, and the specific antibody mixture of a variety of cell adhesion factors is spotted onto the solid-phase carrier at a temperature of 20-26 DEG C under humidity of 35-45%. According to the present invention, with the kit, a variety of the cell adhesion factors commonly used in clinic can be detected, the defects of complex operation, single detection index, low sensitivity and the like in the prior art are overcome, and the kit has advantages of low cost, convenience, high sensitivity, accuracy, high throughput, less sample consumption, promotion and scalization in general laboratories, and the like.

Description

technical field [0001] The invention relates to the technical field of biological detection, in particular to an antibody chip kit for detecting cell adhesion molecules. Background technique [0002] Cell adhesion molecules (CAM) are a general term for many molecules that mediate the contact and binding between cells or between cells and extracellular matrix (ECM). Play a role in the form of receptor and ligand binding. Adhesion molecules enable cell-to-cell, cell-to-matrix adhesion, participate in cell recognition, activation and signal transduction, cell proliferation and differentiation, cell extension and movement, immune response, inflammation, coagulation, tumor metastasis, The molecular basis of a series of important physiological and pathological processes such as wound healing. [0003] Cell adhesion molecules not only have a variety of physiological functions, but also are closely related to the occurrence of pathological processes under certain conditions. Unde...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68G01N33/543
CPCG01N33/54306G01N33/68G01N2333/70525
Inventor 毛应清黄若磐
Owner RAYBIOTECH INC GUANGZHOU
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