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Protein chip and its preparing method and use

A protein chip, protein technology, applied in material testing products, measuring devices, instruments, etc., can solve the problems of protein instability, inability to form products, and difficult to store chips, to solve the problems of degradation over time, avoid instability, and solve Effects that are not easy to save

Inactive Publication Date: 2004-07-14
陈 正豪
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to overcome the instability of the protein itself, which must be repeatedly debugged under strictly controlled conditions. More importantly, once the protein chip is made, the chip is not only difficult to preserve (<7 days), but also cannot be transported. Not to mention the defect of not being able to form a product; and to overcome the defect of the traditional preparation method that must first clone the gene expression fragment, go through the complicated steps of intracellular expression and purification of the synthesized protein; thus providing a long-preserved, practical and commercialized protein chip and its application

Method used

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  • Protein chip and its preparing method and use
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  • Protein chip and its preparing method and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1: Preparation of a protein chip on a high-density island-shaped protrusion glass substrate

[0041] Step 1: treatment and activation of the substrate surface (see figure 2 shown)

[0042] 1) Rinse the graphite substrate with high-density island-shaped protrusions three times with pure water, and then rinse the glass slide three times with 100% ethanol.

[0043] 2) Place the graphite substrate to be plated on the sample stage in the vacuum evaporation table, place the metal to be plated in the evaporation boat, and place the plated metal at 10 under the melting point of gold. -2 Pa plated gold on the surface of the graphite substrate.

[0044] 3) The coated graphite substrate was placed in a layer containing 10-mercaptodecane-NTA (Ca 2+ ) solution, react at room temperature for more than three hours to obtain a substrate connected with an affinity layer.

[0045] 4) Wash the substrate obtained in the above 3) three times with phosphate buffered saline (PBS)...

Embodiment 2

[0058] Example 2: Preparation of protein chips on high-density microporous substrates

[0059] Step 1: Treatment and activation of the chip surface

[0060] 1) Rinse the high-density microporous glass substrate (the cavity is a circular hole with a diameter of 7 microns) three times with water, and then rinse the glass slide three times with 100% ethanol.

[0061] 2) Soak the glass substrate in 0.2M NaOH solution overnight, then wash it three times with high-purity water, and then rinse it once with absolute ethanol.

[0062] 3) Place the cleaned glass substrate in a place containing (CH 3 O) 3 -Si-(CH 2 ) 20 - React in a biotin solution for more than three hours at room temperature to obtain a substrate connected with an affinity layer.

[0063] 4) Wash the substrate obtained in the above 3) three to five times on a shaker with phosphate buffered saline (PBS).

[0064] 5) Wash five times with 70% ethanol, then three times with 95% ethanol, and finally three times with 1...

Embodiment 3

[0072]Example 3: Preparation of Protein Chips on Smooth Substrates

[0073] Step 1: Treatment and activation of the chip surface

[0074] 1) Rinse the smooth quartz substrate three times with water, and then rinse the glass slide three times with 100% ethanol.

[0075] 2) The quartz substrate was soaked overnight in 0.2M HCl solution, then washed three times with high-purity water, and then rinsed once with absolute ethanol.

[0076] 3) Place the cleaned quartz substrate in a place containing (CH 3 O)-Si-(CH 2 ) 22 - Reacting in a solution of glutathione at room temperature for more than three hours to obtain a substrate connected with an affinity layer.

[0077] 4) Wash the substrate in the above 3) with phosphate buffered saline (PBS) on a shaker for a total of three to five times.

[0078] 5) Wash five times with 70% ethanol, then three times with 95% ethanol, and finally three times with 100% ethanol. The prepared substrate connected with the affinity layer is stored...

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Abstract

The present invention relates to marketable high-density protein chip in completely new concept and comprising substrate layer, specific affinity layer and biological macromolecular layer. The protein chip may have protein, antibody or artificial short peptide dots over 1000-100000 in 10 sq cm. The present invention also relates to the process of forming high density precursor protein chip with gene transcription segment capable of performing in-situ geen transcription and translation with chip preparing instrument, and then preparing high density protein chip with no-cell protein expression system. The present invention promotes the practical use and commercialization of protein chip technology and will find its wide application in clinical diagnosis, medical research, medicine development and inspection, protein block research and other fields.

Description

technical field [0001] The present invention relates to a protein chip and its preparation method and application, especially to the protein chip prepared by a cell-free one-step in vitro protein synthesis method and its application in the fields of clinical diagnosis, medical research, new drug development and testing, proteome research, etc. related applications. Background technique [0002] In the context of the determination of the human genome DNA sequence and the gradual and rapid determination of the genomes of a large number of other species, it is an urgent task to develop a set of efficient, fast and practical methods or techniques to study the functions of proteins encoded by the genome in order to obtain A comprehensive and complete understanding of the various biological processes and pathways within the cell. As an efficient, comprehensive and fast method, microarray technology has played its unique role in rapidly obtaining large amounts of parallel data in ...

Claims

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Application Information

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IPC IPC(8): G01N33/50G01N33/543G01N33/68
Inventor 陈正豪朱衡
Owner 陈 正豪
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