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Protein chip based on labeling streptavidin-biotin technology

A streptavidin and protein chip technology, which is applied in the field of life sciences, can solve the problems of affecting the binding efficiency of antigens and antibodies, the inability to detect antibodies, and the limitation of application range, etc., and achieve the effect of improving the detectable range, improving sensitivity, and easy operation

Inactive Publication Date: 2002-02-13
GENE TECH SHANGHAI COMPANY
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  • Abstract
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Problems solved by technology

During the reaction process, due to some non-specific binding, there will be some interference signals in the background. In the case of high sample concentration or poor condition control, false positive reactions may also occur.
In addition, because fluorescent markers are difficult to label on acidic proteins, their scope of application is limited. For example, some antigens are acidic proteins, and the efficiency of fluorescent markers to label antigens is extremely low, so it is impossible to directly use fluorescently labeled antigens to detect antibodies.
However, due to the steric hindrance effect of the molecule, the enzyme label will affect the binding efficiency of the antigen and antibody.

Method used

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  • Protein chip based on labeling streptavidin-biotin technology
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  • Protein chip based on labeling streptavidin-biotin technology

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Embodiment Construction

[0045] The invention is a protein chip using the technology of labeled streptavidin-biotin, and the solid phase carrier of the protein chip can be glass slide, nitrocellulose membrane (NC membrane) or nylon membrane.

[0046] The protein spotting and preparation technique takes the following steps:

[0047] (1) Use a high-speed spotting robot to spot-print protein sample antigens or antibodies on glutaraldehyde-treated glass slides or nitrocellulose or nylon membranes.

[0048] (2) Incubate overnight at room temperature or at 37°C for 1 hour to immobilize on a solid-phase carrier.

[0049] (3) Add a blocking solution based on bovine serum albumin (BSA) and Tween on the surface of the slide, and incubate at 37°C for 1 hour.

[0050] (4) Wash thoroughly with double distilled water and dry at room temperature.

[0051] The hybridization reaction takes the following steps:

[0052] (1) Add the sample to be tested dropwise on the spotted chip, and incubate at 37°C for 1 hour to ...

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Abstract

The present invention relates to protein chip based on labeling streptavidin-biotin technology. The hybridization reaction includes the following steps: dropping tested sample to sample applied chip and washing out redundant sample with elution liquid; blocking wtih blockage liquid, washing and air drying; dropping protein marked with biotin diluted by blockage liquid, washing and air drying; anddropping fluorescnet or enzyme labeled streptavidin diluted by blockage liquid, washing and air drying. The protein chip of the present invention is used in determining antigen and antibody, and detecting specific matter combined with protein. The improved method of the present invention has clear, stable and reliable test results and wide application range.

Description

technical field [0001] The invention belongs to the field of life sciences, in particular to a protein chip using fluorescence or enzyme labeled streptavidin-biotin technology (Fluorescence / Enzyme labeled Streptavidin Biotin Method, FLSAB / ELSAB). Background technique [0002] Labeled streptavidin-biotin technology is a new biological reaction amplification system developed rapidly in recent years, which can be applied to immunoassay. [0003] Protein chips using conventional immunolabeling technology, fluorescent or enzymes are directly labeled on proteins that can specifically bind to the substance to be tested, the reaction results have not been further amplified, the detection sensitivity is limited, and there are high requirements for equipment, reagents and personnel operations. . During the reaction process, due to some non-specific binding, there will be some interference signals in the background. In the case of high sample concentration or poor condition control, f...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K17/14G01N33/543G01N33/68
Inventor 张涛李宾彭永济葛海鹏任一萍
Owner GENE TECH SHANGHAI COMPANY
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