Filtration-based microarray chip

A filter-type, chip-based technology, applied in the field of system composition for detecting specific proteins, can solve problems such as instability of the activation layer

Inactive Publication Date: 2005-06-15
包刚 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the silicon chip needs to be chemically treated and activated before it can bind to the protein, and its activation layer is often unstable.

Method used

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Examples

Experimental program
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Effect test

Embodiment 1

[0117] Assay specificity, sensitivity, reaction kinetics and dynamic range

[0118] We experimentally investigated the following issues to characterize the main features of the filter chip technology of the present invention, including the ability of cellulose membranes to bind fluorescently-labeled and unlabeled proteins. We compared cellulose membrane and glass slide-based protein chips and demonstrated the advantages of filter-based protein chips over shaking-based chips, including faster reaction speed, extended dynamic range, reduced background and improved detection specificity. We also demonstrate the specificity and uniformity of the reaction of a high-throughput multi-chip overlapping filtration system, and the use of filtration technology to enhance the application of phycobilisomes and quantum microparticles. Finally, we demonstrate the filter-type double-antibody sandwich technology and its potential clinical applications.

[0119] Mater...

Embodiment 2

[0184] Double Antibody Sandwich-Filter Chip Technology

[0185] The present invention shows that compared with directly labeling antigens, the double-antibody sandwich method has higher specificity and sensitivity. In addition, the filter-type double-antibody sandwich method is also better than the double-antibody sandwich method in the shaking mode. Combining the use of CEA double-antibody sandwich method and filter-type chip technology, we can effectively detect 5 ng / ml of CEA in blood.

[0186] experimental method

[0187] The protein chip can realize the combined detection of multiple antigens in the patient's blood. The characteristic concentration of carcinoembryonic antigen selected in this case in clinical diagnosis is 5-10 ng / ml, which is 1 / 10 of the total protein concentration in blood 6 . In order to specifically detect carcinoembryonic antigen in blood by direct labeling protein method, the binding ability of carcinoembryonic antigen...

Embodiment 3

[0199] Example 3. Detection of thrombin with filter-type aptamer chip

[0200] Aptamers are a family of single-stranded oligonucleotides that can bind to different molecules after screening. Its binding objects include small organic molecules, peptides and proteins. Compared with antibodies, aptamers are easier to synthesize. A variety of aptamers have been developed, one of which has a high binding affinity for thrombin. In this study, we used this thrombin-specific aptamer as a model system to show that a filter-type aptamer chip can detect fluorescently labeled thrombin. In particular, we investigated the specificity, sensitivity and kinetic range of the technique and compared the reaction kinetics of the filtering and oscillating aptamer chips.

[0201] Materials and methods

[0202] The aptamer specific to thrombin was pre-labeled with biotin (5'-biotin-6 carbon-GGTTGGTGTGGTTGG, purchased from Integrated Gene Company), and then mixed with neostreptomycin and printed on...

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Abstract

The invention proposes a novel filtering protein chip technology. The chip includes a porous cellulose filter carrier, and a microarray formed on the surface of the carrier by a plurality of capture molecules specific to the analyte. Capture molecules can be proteins (including monoclonal or polyclonal antibodies, antigens or other proteins), aptamers or other small molecules. In addition, the present invention also includes a device and an experimental method for simultaneously using multiple stacked microarray chips to detect analytes. In short, the filter-type protein chip is a new technology that is more sensitive, more specific, more quantitative, faster and higher throughput, and can be used for many applications such as multi-molecule joint detection of diseases.

Description

field of invention [0001] The invention relates to a system composition and an experimental method for detecting specific proteins in biological samples. Specifically, microarrays composed of capture molecules are used to achieve high-throughput detection of samples to be analyzed, such as protein molecules specific to diseases. Background technique [0002] As an emerging field of life sciences, proteomics is based on genome research and is an important development of the latter. The proteome is all the encoded proteins of a genome, and proteomics is "the study of gene expression at the protein level, in order to deeply study life processes (such as disease mechanisms and drug efficacy) and decode the control laws of gene expression". Genomics has provided preliminary knowledge about the impact of gene expression and regulation on some diseases and drug treatments. Emerging technologies, including sequence analysis of gene expression (SAGE) and microarrays (which have bee...

Claims

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Application Information

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IPC IPC(8): B01J19/00B01L3/00B01L9/00C40B40/06C40B40/10C40B40/12C40B60/14G01N1/28G01N1/40G01N33/543G01N33/68
CPCB01J19/0046B01J2219/00286B01J2219/00378B01J2219/00527B01J2219/00585B01J2219/00596B01J2219/00641B01J2219/00659B01J2219/00668B01J2219/00677B01J2219/00707B01J2219/00722B01J2219/00725B01J2219/00731B01L3/50255B01L9/523B01L2300/0681B01L2300/0819B01L2300/0877B82Y30/00C40B40/06C40B40/10C40B40/12C40B60/14G01N1/40G01N1/4077G01N33/54366G01N33/54386G01N33/6803
Inventor 包刚许扬清
Owner 包刚
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