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Cell-based biosensor used for detection of bacterial-colony signal molecules and preparation method and application of cell-based biosensor

A technology of cell sensors and signaling molecules, applied in the direction of material electrochemical variables, material resistance, etc., to achieve the effects of improving adhesion, increasing reaction speed, and improving sensitivity

Inactive Publication Date: 2017-07-21
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There have been many reports on cell sensors, but there is no precedent for using cell sensors to detect AHLs

Method used

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  • Cell-based biosensor used for detection of bacterial-colony signal molecules and preparation method and application of cell-based biosensor
  • Cell-based biosensor used for detection of bacterial-colony signal molecules and preparation method and application of cell-based biosensor
  • Cell-based biosensor used for detection of bacterial-colony signal molecules and preparation method and application of cell-based biosensor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Preparation of electrochemical cell sensor

[0036] 1. Preparation of graphene oxide and sodium alginate / graphene oxide mixed solution configuration

[0037] Graphene oxide is prepared from natural graphene. Put graphene in a 250mL flask, then add concentrated sulfuric acid, sodium nitrate, potassium permanganate and stir, maintain the temperature at 20℃, and stir for 2h; add distilled water and raise the temperature to 35℃. Keep for 0.5h; heat to 90℃, keep for 0.5h; add 30% H at last 2 O 2 , Until the color becomes bright yellow and no gas is produced. The mixture was filtered and washed with 5% HCl and distilled water for 3 times, the filter residue was dried and dissolved in water, ultrasonicated for 3 hours to form a uniform dispersion, and dried again to obtain graphene oxide. Mix 0.1 g of graphene oxide with 100 mL of RPMI 1640 culture solution, and ultrasonically treat it for 5 minutes to obtain a uniform dispersion. Add 10% (wt%) graphene oxide mixture dro...

Embodiment 2

[0042] Example 2 Application of electrochemical cell sensor

[0043] 1. Standard test

[0044] Weigh 3OC of different quality 12 -HSL standard product, dissolved in DMSO. RBL-2H3 cells were cultured to the logarithmic growth phase, digested and centrifuged, collected and counted, and the cell concentration was controlled at 10 7 cells / mL. Pipette 5μL of cell suspension into a small centrifuge tube, and add 3OC with final concentration of 0.01μM, 0.25μM, 0.5μM, 0.75μM, 1μM, 2.5μM, 5μM, 7.5μM, 10μM to each well. 12 -HSL solution, 2 wells in parallel, placed in a cell incubator and incubated for 30 minutes. Take out the cell sensor electrode that has been cleaned and polished, and add 5μL of 3OC with different concentrations in turn 12 -The cell suspension stimulated by HSL standard, covered with sodium alginate / graphene oxide mixture, fixed in CaCl2 solution for 1 min and incubated at 37°C for 5 min, then insert the electrode into the electrolytic cell for measurement.

[0045] 2. An...

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Abstract

The invention discloses a cell-based biosensor used for detection of bacterial-colony signal molecules and a preparation method of the cell-based biosensor. Graphene oxide is added to a sodium alginate solution to form a sodium alginate / graphene oxide mixture, a cell suspension solution is inoculated to the surface of a gold electrode, cells are covered and wrapped with the sodium alginate / graphene oxide mixture and are cultivated and inoculated after soaked in a CaCl2 insulation to form hydrogel to obtain the cell-based biosensor; the constructed cell-based biosensor is applied to detection of 3OC12-HSL of N-acyl-homoserine lactone (AHLs) standard substances in the bacterial-colony signal molecules, good linear relation of the concentration at 0.01mu M to 1um M is achieved, and detection limit is 0.01um M. Identification, conduction and detection on the standard substances of the colony signal molecules are realized accurately, efficiently and rapidly, shortcomings in traditional detection methods are overcome, and the cell-based biosensor is simple in manufacture, low in cost and good in application prospect.

Description

Technical field [0001] The invention relates to an electrochemical sodium alginate / graphene oxide hydrogel cell sensor and a preparation method thereof. The invention belongs to 3OC 12 -HSL and other bacterial population signal molecular detection technology fields. Background technique [0002] For a long time, people believed that bacteria lived in the environment as a single cell. There would be no communication between bacteria and bacteria, unable to perceive changes in the same population or other populations, and there would be no spontaneous group behavior. However, studies in the past 30 years have shown that there is information exchange between bacteria and bacteria, and the medium of communication is a small molecule substance, which we call "signal molecules." Bacteria and bacteria perceive the quantitative changes of their own populations through signal molecules. When the population reaches a certain number or concentration, the signal molecules facilitate the bind...

Claims

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Application Information

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IPC IPC(8): G01N27/30G01N27/06G01N27/07
CPCG01N27/30G01N27/06G01N27/07
Inventor 方维明蒋栋磊尹永祺刘延葛庆丰徐鑫
Owner YANGZHOU UNIV
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