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Protein chip carrier processing method

A protein chip and solid-phase carrier technology, applied in the field of protein chips or immunoassays, can solve the problems of signal interaction at different points, high background, low sensitivity, etc.

Inactive Publication Date: 2007-09-19
穆海东
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] (1) The sensitivity is low,
[0013] (2) The background is high,
[0014] (3) The protein lattice is easy to diffuse during the reaction process, especially when the density of the array is large, it is easy to cause the signals of different points to affect each other due to the close distance

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] In this example, a protein chip was prepared by taking the detection of five kinds of antibodies against hepatitis B, including anti-HCV, anti-Core, anti-NS3, anti-NS4, and anti-NS5, as an example.

[0050] (a) Preparation of protein chip solid phase carrier

[0051] Choose a hydrophilic NC membrane with a pore size of 0.4mm produced by Whatman, soak it in 100ug / ml avidin-biotin complex solution with biotin accounting for 1 / 4 of the active unit of avidin at room temperature for 120 minutes at room temperature, pH Wash with 7.8 PBS buffer 3 times, and dry at room temperature. The avidin used in the complex is streptavidin and the biotin is long-chain biotin.

[0052] (b) Preparation of protein chip

[0053] Use a high-speed protein spotting instrument to spot the corresponding five antigens of the five hepatitis C antibodies on the above-mentioned avidin-biotin-treated NC membrane and on the untreated NC membrane, each array 16 (4 × 4 ) samples. It was then blocked wit...

Embodiment 2

[0058] The sequencing of Example 1 was repeated, the only difference being that different concentrations or ratios of avidin-biotin complexes and different solid phase carriers were used (see Table 2 for details).

[0059] Solid phase carrier

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PUM

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Abstract

The present invention provides one kind of solid carrier for protein chip with the compound of avidin and biotin combined to the surface, with the number of the biotin in the compound being 0.15-0.4 active avidin unit. The solid carrier for protein chip has obviously lowered diffusion of protein sampling point and raised detection sensitivity. The present invention also provides the preparation of the solid carrier for protein chip.

Description

technical field [0001] The invention belongs to the field of protein chip or immunoassay, and more specifically relates to a new protein chip carrier and a processing method thereof. Background technique [0002] In order to improve the detection throughput and detection efficiency, a microarray enzyme-linked immunoassay technology, also known as protein chip technology, has been developed in recent years. It uses the specificity of antibody-antigen binding, that is, the immune response to detect samples. [0003] The simplified model of protein chip construction is: select a solid phase carrier that can firmly bind protein molecules (antigens or antibodies), thus forming a protein microarray, that is, a protein chip. The protein microarray is constructed on the substrate, and each point in the array represents an independent biochemical reaction, so that many reactions are integrated on a small carrier. [0004] According to Rou-Pan huang, Thomas O.joos, etc., the construc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/53G01N33/68G01N33/543
Inventor 穆海东
Owner 穆海东