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Liquid biochip system

A liquid chip and chip technology, applied in biological testing, material inspection products, instruments, etc., can solve the problems of small amount of modified samples, insufficient chip bonding, and inability to ensure full chip bonding, etc.

Inactive Publication Date: 2009-12-02
毅新兴业(北京)科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This type of chip has its inherent disadvantages: 1. Chip specificity, which is related to chip surface modification and modifiers. Poor chip surface modification process or improper spacing of modifiers may cause non-specific adsorption on the chip surface. The specificity of the modifier directly determines the specificity of the chip
2. Chip capacity, the surface area of ​​the chip is limited, so the number of modifiers to be modified is certain, therefore, the chip capacity has always been an important issue for the chip
3. The combination of the chip and the sample is insufficient. The combination of the chip and the molecule to be tested is a planar combination, and the amount of sample required by the chip is very small, so it cannot be guaranteed that the chip is fully combined with the sample. This is not like the two items in the liquid can be fully mixed. ;4. Insufficient elution of the chip. The chip is a solid surface. The elution cannot be completely washed, which may cause incomplete washing of interfering substances and affect the specificity of the chip; 5. The amount of modified samples is small, and the detection of low-abundance target substances limited ability
[0004] These problems are mainly limited to the inability of the modified surface to mix well with the sample.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Embodiment one: the preparation of microbead

[0074] 1. Chitosan Microbeads

[0075] Chitosan (chitosan), chemical name (1,4)-A-amino-A-deoxy-B-D-glucan, is a copolymer of N-deacetylglucosamine, because chitosan and its derivatives are free of Toxicity, biodegradability and good biocompatibility, it is widely used as immobilized carrier of enzymes and cells.

[0076] The following is the preparation process of chitosan microbeads: weigh chitosan 9, dissolve it in 300mL of 2% acetic acid solution, stir continuously for 2 hours to fully dissolve, remove insoluble matter by suction filtration, add Tween-80 10mL, oil Phase dispersion medium, porogen ethyl acetate, emulsifier and polymer surface modifier, fully stirred and reacted at 50°C for 30 minutes, raised the temperature to 60°C, added formaldehyde solution, reacted for 30 minutes, then added 2.0ml of glutaraldehyde, Adjust the pH to 9.0, raise the temperature to 80°C, and react for 60 minutes. Suction filter, wash ...

Embodiment 2

[0102] Example 2: Surface functional group modification of microbeads

[0103] 1. Surface affinity modification process of chitosan microbeads

[0104] Add appropriate amount of deionized water to chitosan microbeads, activate with epichlorohydrin, add EDC and heparin, and stir at 4°C for 24 hours to obtain surface affinity-modified microbeads.

[0105] The long-chain alkyl group of chitosan microbeads is used as the hydrophobic part, and the sulfate group is used as the hydrophilic part to synthesize N-octyl-O-sulfate chitosan (OCS1) to obtain microbeads with surface hydrophilic modification.

[0106] 2. Surface ion exchange modification process of cellulose microbeads

[0107] Add the washed cellulose microbeads to a solution containing 0.1mol / L DEAE hydrochloride and 0.5mol / L NaOH, and keep stirring at 60°C for 10 hours to make it cross-link-DEAE weakly basic anion exchange group group.

[0108] 3. Metal chelation affinity modification of microbeads

[0109] Add microbe...

Embodiment 3

[0115] Example 3: Binding, purification and identification of acidic protein and metal binding protein

[0116] (1) Take 98ul of binding buffer in a 0.5ml eppendorf tube and add 2ul of serum. Select a weak cation binding separation device, fix the separation device at the front end of a 20-200ul pipette, draw the sample in the EP tube, gently pipette back and forth 5 times, and let it stand for 1min. Blow off the sample. Add 0.6ml of washing buffer into the 1.5ml EP tube, adjust the pipette to 120ul to absorb the washing buffer, pipette in another EP tube three times, discard the washing buffer, and repeat this three times. Aspirate 20ul of elution buffer and let stand for 2mins. Aspirate the eluate into a new 0.2ul EP tube. The eluate can be detected and identified by protein spectrum, and the results are as follows: figure 2 -A, wherein the abscissa represents the mass-to-charge ratio of the detected protein (wherein the charge is 1), and the ordinate is the peak intens...

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Abstract

The invention discloses a liquid biochip system for binding, separating and purifying a targeting biomacromolecule. The system comprises a liquid chip consisting of a micron or nanometer chip particle and chip particle storage buffer solution, and liquid chip work buffer solution, wherein the liquid chip work buffer solution comprises sample modifying buffer solution, binding buffer solution, washing buffer solution, eluting buffer solution, and / or chip passivating buffer solution, and / or chip activating buffer solution. The system has the advantages of good repeatability, high sample processing throughput, simple operation, low cost and the like, has extremely wide application field, and is applicable to separation and purification of various biomacromolecules.

Description

technical field [0001] The invention relates to the diagnosis and biochip technology of biomacromolecules in proteomics and molecular biology, and specifically provides a liquid biochip system to absorb, separate, purify and detect target biomacromolecules in biological samples. technical background [0002] Biochip is a high-tech developed rapidly in the field of life sciences in the late 1980s. It mainly refers to a micro-biochemical analysis system built on the surface of a solid chip through micro-processing technology and microelectronics technology to realize the analysis of living organisms. Accurate, rapid and informative detection of tissues, cells, proteins, nucleic acids, carbohydrates and other biological components. Biochip technology has the characteristics of high throughput, miniaturization and automation. Using biochip technology, multiple indicators can be tested on the detected object at one time. The highly integrated tens of thousands of densely arrang...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/48G01N33/53G01N33/547G01N21/64C08L5/08C08L83/06C08L67/04C08L27/16C08L29/04
Inventor 马庆伟于中连吕萍萍任晶胡晓慧张燕
Owner 毅新兴业(北京)科技有限公司
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