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: 2015-05-27
RAYBIOTECH INC GUANGZHOU
View PDF2 Cites 9 Cited by
  • 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

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Antibody chip kit for simultaneous quantitative detection of multiple receptors
  • Antibody chip kit for simultaneous quantitative detection of multiple receptors
  • 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 a carrier. Since the nitrocellulose membrane is a multi-layer structure, it is difficult to wash the chip, resulting in 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. Different manufacturers use different methods to immobilize nitrocellulose membranes on the surface of glass slides. Among them are Whatman's SS slides, which fix 16 cells containing nitrocellulose membranes on one glass slide. In addition, Gentel uses processed PATH slides were produced by coating the slide surface with nitrocellulose material. In addition, in order to immobilize the antibody on the surface of the glass slide, we screened the carriers activated by different methods. Use a fully automati...

Embodiment 2

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

[0029] In order to detect whether there are corresponding receptors in the sample, prepare slides immobilized with specific antibodies against the following proteins: tumor necrosis factor receptor 4-1BB, leukocyte-activated adhesion factor, stimulatory molecule B7-1, B cell maturation factor , myeloid cell-specific leucine-rich 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 kinases, cell membrane glycoproteins, receptor 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-associated protein, herpes virus entry mediator, intercellular adhesion molecule 3, interleukin-1...

Embodiment 3

[0047] Example 3: Experiment of 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, peel off 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 serial dilutions of cytokine standard gradients as shown in

[0051] 2.1. Add 500 μl of sample diluent to the vial of the cytokine 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.

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

[0053] 2.3. Take 100 μl of Std1 and add it to Std2 and mix gently, then take 100 μl from Std2 and add it t...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

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 invention relates to the technical field of biomedicine, in particular to an antibody chip kit for quantitatively detecting multiple receptors at the same time. technical background [0002] A receptor is a protein that can bind to a specific extracellular signaling molecule (ligand) to trigger a cellular response. [0003] Receptors are divided into cell surface receptors and intracellular receptors. When the receptor binds to the ligand, the molecular conformation changes, thereby causing cellular responses, such as mediating cellular processes such as intercellular signal transduction, intercellular adhesion, and cell endocytosis. Receptors can recognize bioactive molecules on the cell membrane or in the cell and bind to them, amplify and transmit the recognized and received signals to the inside of the cell, thereby activating or starting a series of biochemical reactions, and finally lead to the specificity of the signal substance. biologica...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/68
CPCG01N33/54306G01N33/68G01N2333/705
Inventor 毛应清黄若磐
Owner RAYBIOTECH INC GUANGZHOU
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap