One-dimensional biological chip and application in gene, protein expression analysis

A protein expression and biological maintenance technology, applied in the field of microfluidic-based gene or protein string analysis technology, can solve the problems of crossover and fusion

Inactive Publication Date: 2005-07-06
HUNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Judging from the current literature reports, these two chip technologies have their own technical advantages and research fields. Although they are

Method used

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  • One-dimensional biological chip and application in gene, protein expression analysis
  • One-dimensional biological chip and application in gene, protein expression analysis
  • One-dimensional biological chip and application in gene, protein expression analysis

Examples

Experimental program
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Effect test

Embodiment 1

[0026] Example 1 (Construction of a one-dimensional biochip and its application in the study of cellular protein expression profiles)

[0027] Monoclonal antibody modified on the surface of organic polymer particles: Take about 10 mg of polystyrene particles with a diameter of 40 μm and place them in a 0.5 ml Ep tube, add 100 μl of 20 μg / ml mouse anti-monoclonal antibody, and place on a low-temperature shaker (4°C, >300rpm) for shaking 24h. Wash 3 times with 10 mM PBS buffer, add 10 mM PBS buffer containing 0.1%-3% BSA, place on a low-temperature shaker (4°C, >300 rpm) and shake for 12 hours. Take it out and store it at 4°C for later use.

[0028] Pouring the PDMS substrate of the one-dimensional biochip with polydimethylsiloxane (PDMS) on the prepared positive template, placed in an oven at 75°C for about 40 minutes, took it out after curing, and removed the PDMS substrate from the positive template. Peel off. Place the PDMS film base under a Leica inverted microscope, and...

Embodiment 2

[0030] Example 2 (one-dimensional biochip detection of p53 protein expression in tumor cell CNE2):

[0031] Monoclonal antibody modified on the surface of aminated silica particles: Take about 10 mg of aminated silica particles with a diameter of 40 μm and place them in a 0.5 ml Ep tube, add 100 μl of 4% glutaraldehyde, and place on a low-temperature shaker (4°C, >300rpm ) shaken for 2 hours, washed with secondary water for 3 times; added 100 μl of 20 μg / ml mouse anti-P53 monoclonal antibody, placed on a low-temperature shaker (4°C, >300 rpm) and shaken for 12 hours. The reacted particles were washed three times with 10 mM PBS buffer, added with 10 mM PBS buffer containing 0.1%-3% BSA, and placed on a low-temperature shaker (4° C., >300 rpm) for 12 h. Take it out and store it at 4°C for later use.

[0032] Pouring the PDMS substrate of the one-dimensional biochip with polydimethylsiloxane (PDMS) on the prepared positive template, placed in an oven at 75°C for about 40 minutes...

Embodiment 3

[0034] Embodiment 3 (one-dimensional biochip detects the expression of p53 protein in several kinds of cells)

[0035] Monoclonal antibody modified on the surface of silica particles: Take about 10 mg of silica particles with a diameter of 40 μm and place them in a 0.5 ml Ep tube, add 200 μl of 2M Na 2 C0 3 The solution was activated for 15-30 minutes, and then 100 μl of 1 g / ml CNBr acetonitrile solution was added, and the reaction was continued for 30 minutes. The reacted particles were fully washed 3 times with ice water and 3 times with 10 mM PBS buffer. Add 100 μl of 20 μg / ml mouse anti-P53 monoclonal antibody to the Ep tube, and place on a low-temperature shaker (4° C., >300 rpm) to shake for 24 hours. The reacted particles were washed three times with 10 mM PBS buffer, added with 10 mM PBS buffer containing 0.1%-3% BSA, and placed on a low-temperature shaker (4° C., >300 rpm) for 12 h. Take it out and store it at 4°C for later use.

[0036] Pouring the PDMS substrate...

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Abstract

Disclosed is an one-dimensional biochip belonging to a serial analytic technique for micro-total analysis systems. The one-dimensional biochip is provided with a plurality of cellules on the micro-separation channel of micro flow control chip, microparticles modified by different biomolecules on surface are arranged in different cellules; in the gene or protein analysis, when the sample flows through the microchannel with a plurality of cellules, the microparticles can specifically identify and capture multiple target molecules, then a reagent with fluorescence labels can be introduced, and finally the microparticles will specifically combine with the fluorescence labels on surface to be detected by fluorescent imaging. The one-dimensional biochip provided by the invention not only has advantages of micro flow control technology and array analysis, but also improves the detection sensitivity and identification capability of target molecules, and provides an effective research measure for the gene and protein expression analysis at single cell level, and tumor research and drug screening.

Description

Technical field: [0001] The invention relates to the field of micro-total analysis systems, in particular to a microfluidic-based gene or protein string analysis technology. Background technique [0002] Micro total analysis system (μ-TAS, also known as lab-on-chip, Lab-on-chip) has become one of the fastest-growing important frontiers in the field of life science research. The micro-analysis system with the characteristics of miniaturization, integration, multi-function and high-throughput makes it possible to systematically study the gene expression and protein expression of a biological system, and will revolutionize biomedical testing and disease diagnosis. At present, chip technology at home and abroad is generally developing in two directions: one is microfluidic chips, which are mainly based on analytical chemistry and analytical biochemistry, relying on micro-electromechanical processing technology, and characterized by micro-pipeline networks. The focus of the deve...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/68
Inventor 王柯敏周雷激谭蔚泓左新兵文建辉陈韵晴张何
Owner HUNAN UNIV
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