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Antibody chip kit for screening biomarkers

A biomarker and antibody chip technology, applied in the field of biomedicine, can solve the problems of low sensitivity, cumbersome operation, expensive instruments, etc., achieve high-throughput screening, overcome cumbersome operation, and use less specimens

Active Publication Date: 2015-04-29
RAYBIOTECH INC GUANGZHOU
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] Aiming at the deficiencies of the prior art, the technical problem to be solved by the present invention is to provide a high-throughput, high-sensitivity, high-specificity and low-cost cytokine detection kit, which can simultaneously detect sixty cell factors by using protein chip technology. factor, which overcomes the shortcomings of the existing technology such as cumbersome operation, single detection index, need for expensive instruments, and low sensitivity. advantages

Method used

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  • Antibody chip kit for screening biomarkers
  • Antibody chip kit for screening biomarkers
  • Antibody chip kit for screening biomarkers

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1: Preparation of an antibody chip kit for screening biomarkers.

[0019] In order to screen the corresponding biomarkers in patient specimens, the base membrane immobilized with specific antibodies against the following proteins: angiopoietin, brain-derived neurotrophic factor, B-lymphocyte chemotactic factor, bone morphogenetic protein- 4. Bone morphogenetic protein-6, CKβ8-1, ciliary neurotrophic factor, epidermal growth factor, chemokine, chemokine-2, chemokine-3, fibroblast growth factor-6, fibroblast Growth factor-7, FMS-like tyrosine kinase 3 ligand, chemokine CX3CL1, chemokine (C-C motif) ligand 6, glial cell-derived neurotrophic factor, granulocyte colony-stimulating factor, chemokines (C-C motif) ligand 1, γ-interferon, insulin-like growth factor binding protein-1, insulin-like growth factor binding protein-2, insulin-like growth factor binding protein-4, insulin-like growth factor 1, interleukin -10, Interleukin-13, Interleukin-15, Interleukin-16, In...

Embodiment 2

[0033] Embodiment 2: Experiment of detecting cytokines with the kit of the present invention.

[0034] Put the bottom film in the supporting square box, since each bottom film in this embodiment is distributed with a chip lattice, so in this embodiment, the square box is provided with 8 squares, and each square is placed with a bottom film , each chip dot matrix becomes a mutually independent reaction area through the grid. The box with 8 grids used in this example was self-made by the inventor. After adding 2 ml of blocking solution to each grid, place it at room temperature and incubate for 30 minutes, and then perform the following steps in sequence:

[0035] 1. Add sample

[0036] Aspirate the blocking solution in each square, put 1-2 ml of the sample diluted with the blocking solution into the square with a membrane, and then shake it on a shaker at room temperature for 1 to 2 hours, or at 4°C Under reaction 12-18 hours.

[0037] 2. Membrane washing

[0038] Washing s...

Embodiment 3

[0053] Example 3: Test of cross-reactivity of the kit of the present invention.

[0054] The cross-reactivity test between antibody pairs was carried out according to the following method: adding the same 60 kinds of cytokine standard proteins to different bottom membranes. After incubation, slides are washed and then incubated with individual detection antibodies for each antigen. Finally, it was incubated with horseradish peroxidase-labeled streptavidin, and the chip was scanned and read. Taking the capture antibody of each antigen as the horizontal axis and the added detection antibody as the vertical axis, the experimental results in Table 2 can be obtained.

[0055] Table 2 Cross-reaction test results between antibody pairs

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[0072] IL-13

[0073] The experimenta...

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Abstract

The invention discloses an antibody chip kit for screening biomarkers. The kit comprises an antibody chip, a biotin labeled cell factor detection antibody mixture and horse radish peroxidase labeled streptavidin, wherein the antibody chip comprises a bas membrane and specific antibodies of 60 cell factors fixed on the surface of the base membrane, three positive control. The kit disclosed by the invention can be used for simultaneously detecting 60 cell factors for multiple samples, overcomes the defects in the prior art of tedious operation, single detection index, low sensitivity and the like and has the advantages that the kit is cheap, convenient, sensitive, accurate, high in throughput and few in use level of specimens and can be popularized and expanded on a common laboratory and the like.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to an antibody chip kit for biomarker screening. Background technique [0002] Cytokines are proteins or small molecular polypeptides that are synthesized and secreted by the body's immune cells and non-immune cells and have immune regulation and effector functions. [0003] Cytokines have a wide range of biological activities, which can promote the proliferation and differentiation of target cells, promote or inhibit the expression of other cytokines and membrane surface molecules, enhance anti-infection and cell killing effects, promote inflammatory processes, and affect cell metabolism. Cytokines can be classified into interleukins, interferons, tumor necrosis factor superfamily, colony-stimulating factors, chemokines, growth factors, and the like. Many cytokines play a role in the body through paracrine, autocrine or endocrine, etc., and have various physiological character...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/6863G01N33/6866G01N33/6869G01N33/6872G01N2333/71G01N2333/715G01N2333/7155G01N2333/7156G01N2333/7158
Inventor 黄若磐
Owner RAYBIOTECH INC GUANGZHOU
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