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An antibody chip kit for simultaneous quantitative detection of multiple receptors

An antibody chip and quantitative detection technology, applied in the field of biomedicine, can solve the problems of cumbersome operation, low throughput, low sensitivity, etc., and achieve the effect of overcoming cumbersome operation and less specimen consumption

Active Publication Date: 2018-08-17
RAYBIOTECH INC GUANGZHOU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, one test can only detect a single indicator, with low throughput and high cost. In the detection of receptors with multi-indicator characteristics, this method has obvious shortcomings; although radioimmunoassay (RIA) has high sensitivity, it is due to radioactive contamination. It is rarely used now; immunoblotting can measure the size of molecules, and there is no non-specific reaction, but the operation is cumbersome, the sensitivity is low, and it can only detect a single indicator, which is not suitable for the detection of receptors; flow cytometry can be used in the detection of receptors. Detect the level of receptors at the cellular level, but there are disadvantages such as low sensitivity, low throughput, and high cost

Method used

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  • An antibody chip kit for simultaneous quantitative detection of multiple receptors
  • An antibody chip kit for simultaneous quantitative detection of multiple receptors
  • An antibody chip kit for simultaneous quantitative detection of multiple receptors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1: Screening of the best antibody chip carrier.

[0025] Conventional antibody chips mostly use nitrocellulose membrane as the carrier. Because the nitrocellulose membrane has a multi-layer structure, it is difficult to clean the chip, so that the background of the chip is high and the result is large. At the same time, the nitrocellulose membrane is fragile and not easy to operate on a large scale, and the use of large-scale clinical samples is not widespread. Different manufacturers use different methods to fix the nitrocellulose membrane on the surface of the glass slide. Among them is Whatman's SS glass slide, which fixes 16 cells containing nitrocellulose membrane on a glass slide. In addition, Gentel has used processing The nitrocellulose material is coated on the surface of the glass slide to produce the PATH glass slide. In addition, in order to fix the antibody on the surface of the glass slide, we screened the carrier activated by different methods. Spo...

Embodiment 2

[0028] Example 2: Preparation of an antibody chip kit for the simultaneous quantitative detection of multiple receptors.

[0029] In order to detect whether there are corresponding receptors in the sample, prepare slides fixed with specific antibodies against the following proteins: tumor necrosis factor receptor 4-1BB, leukocyte activation adhesion factor, stimulating molecule B7-1, B cell maturation factor , Myeloid cells are specifically rich in leucine glycoprotein, tumor necrosis factor receptor CD30, tumor necrosis factor ligand-5, carcinoembryonic antigen-related cell adhesion molecule 1, death receptor 6, Axl / Ufo growth factor receptor Family tyrosine kinase, cell membrane glycoprotein, receptor type tyrosine kinase ErbB3, E-selectin, tumor necrosis factor receptor superfamily member 6, human FMS-like tyrosine kinase 3 ligand, glucocorticoid-induced Tumor necrosis factor receptor family related proteins, herpes virus invasion mediator, intercellular adhesion molecule 3, i...

Embodiment 3

[0047] Example 3: Experiments for quantitative detection of receptors using the kit of the present invention.

[0048] 1. Complete drying of the slide chip

[0049] Take the slide chip out of the box, and after equilibrating at room temperature for 20-30 minutes, open the packaging bag, remove the sealing strip, and then place the chip in a vacuum desiccator or dry at room temperature for 1-2 hours.

[0050] 2. Press image 3 Perform a serial dilution of the cytokine standard gradient as shown in

[0051] 2.1. Add 500μl of sample diluent to the small tube of cytokine standard mixture, and re-dissolve the standard. Before opening the small tube, centrifuge it quickly, gently tap up and down to dissolve the powder, and mark this small tube as Std1.

[0052] 2.2. Mark 6 clean centrifuge tubes as Std2, Std3 to Std7, and add 200μl of sample diluent to each tube.

[0053] 2.3. Extract 100μl of Std1 and add it to Std2 and mix gently, then extract 100μl from Std2 and add it to Std3, so as to g...

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Abstract

The invention discloses an antibody chip kit for simultaneous quantitative detection of multiple receptors. The kit comprises antibody chips, a receptor standards mixture, a biotin labeled detection antibodies mixture and Cy3 fluorochrome labeled streptavidin. A standard tissue slide which is coated by reactive amino groups is chosen as a surface carrier of the antibody chips, and various antigen specific antibodies are adsorbed firmly on the surface of the slide by a non-contact spotting robot and sixteen microarrays are formed. The sixteen microarrays are then separated in non-interfering small areas by the use of a plastic frame with 2*8 holes and a latex closed frame which match each other and are demountable. Each capture antibody has four repeated spottings in each antibody microarray. The antibody chip kit can realize simultaneous quantitative detection of multiple receptors. Dynamic detection range of receptors is wider. Quadruple ELISA repeated data can be obtained from a single sample experiment. The antibody chip kit has advantages of high sensitivity, high throughput, small required use amount of samples, low cost, easy popularization and the like.

Description

Technical field [0001] The present invention relates to the field of biomedical technology, in particular to an antibody chip kit for quantitatively detecting multiple receptors at the same time. technical background [0002] Receptor is a protein that can bind to a specific signal molecule (ligand) outside the cell to cause a cellular response. [0003] Receptors are divided into cell surface receptors and intracellular receptors. The binding of receptors and ligands will cause molecular conformational changes, which will cause cellular reactions, such as mediating cell-cell signal transduction, cell-cell adhesion, cell endocytosis and other cellular processes. The receptor can recognize and bind to bioactive molecules on the cell membrane or in the cell, and correctly amplify and transmit the recognized and received signals to the inside of the cell, thereby activating or initiating a series of biochemical reactions, and finally leading to the specific signal substance Biologic...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68
CPCG01N33/54306G01N33/68G01N2333/705
Inventor 毛应清黄若磐
Owner RAYBIOTECH INC GUANGZHOU
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