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A storage method for paper-based biochemical reagents based on aqueous two-phase system

A two-phase system and biochemical reagent technology, applied in chemical instruments and methods, laboratory containers, laboratory utensils, etc., can solve the problems of inconvenient storage of reagents, and achieve wide application range, good biocompatibility, effective Conducive to the effect of commercial promotion

Active Publication Date: 2022-03-01
HANGZHOU TINKER BIOTECHNOLOGY CO LTD
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Problems solved by technology

[0004] The purpose of the present invention is to provide a simple and reliable method for the stable storage of biochemical reagents that can be used in the field of on-site instant detection, so as to solve the problem of inconvenient storage of reagents in existing paper chips and other portable micro-devices

Method used

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  • A storage method for paper-based biochemical reagents based on aqueous two-phase system
  • A storage method for paper-based biochemical reagents based on aqueous two-phase system
  • A storage method for paper-based biochemical reagents based on aqueous two-phase system

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Effect test

Embodiment 1

[0030] The storage of embodiment 1 horseradish peroxidase in dextran phase

[0031] Horseradish peroxidase was dissolved in 5% (w / v) dextran solution and 10 mM phosphate buffer (pH 7.4), respectively, with a final concentration of 0.05 μg / mL. 3,3',5,5'-Tetramethylbenzidine and aqueous hydrogen peroxide were mixed at a volume ratio of 1:6 to form a substrate solution. Take 10 μL of the above two enzyme solutions in two centrifuge tubes, add 70 μL of substrate solution respectively, react for 15 minutes, add 10 μL of sulfuric acid solution (2M) to stop the reaction, and measure the ultraviolet-visible spectra of the two reaction solutions by spectrophotometry ( figure 1 ). Experiments have shown that dextran in solution does not lead to a decrease in enzyme activity.

[0032] Get four parts of above-mentioned dextran enzyme solutions (every part of 1mL), each part of solution adds different amounts of iron-ethylenediaminetetraacetic acid complex solution, so that the final con...

Embodiment 2

[0034] Example 2 The paper chip storing reagents is used for on-site instant detection of carcinoembryonic antigen

[0035] The detection antibody labeled with horseradish peroxidase is dissolved in a homogeneous polyethylene glycol / dextran solution, and the solvent volatilization induces phase separation to form a two-layer solution, the lower layer is the dextran phase, and the upper layer is the polyethylene glycol phase. The detection antibody labeled with horseradish peroxidase will be selectively distributed in the lower dextran phase during the phase separation process, such as Figure 4 shown. Take out the polyethylene glycol phase and the dextran phase after the phase separation respectively, add the substrate to react, measure the absorption spectrum curve of the reaction solution after the sulfuric acid solution terminates the reaction, as Figure 5 shown. The experiment proved that the detection antibody labeled with horseradish peroxidase was mainly distributed ...

Embodiment 3

[0038] Example 3 Antibody Storage Experiment on a Paper Chip

[0039] Put the antibody-loaded paper chip into a vacuum-sealed bag and store it at 4°C and room temperature (about 25°C), respectively, take out the paper chip at a specific time point, and measure 20ng / mL carcinoembryonic antigen using the above-mentioned carcinoembryonic antigen detection process The Blue / Red value of the paper chip is compared with the Blue / Red value measured by the unpreserved paper chip, and the results are as follows Figure 9 shown. Experiments show that after 104 days of storage at 4°C and at room temperature, the detection effects of the paper chips are maintained at about 90% and 70%, respectively.

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Abstract

A paper-based biochemical reagent storage method based on a water two-phase system, comprising: dissolving the biochemical reagent to be stored in the water two-phase system solution, and then adding the water two-phase system solution dropwise to the reagent storage area of ​​the paper chip, and the solvent volatilizes Induced phase separation in an aqueous two-phase system to form an aqueous bilayer film. The biochemical reagents are selectively distributed in the lower film, and the upper film inhibits the volatilization of water in the lower film, avoiding direct contact between the reagents and the external environment, and improving the storage stability of the biochemical reagents. Afterwards, the reagent-loaded paper chip was sealed in a vacuum-sealed bag and stored at 4°C or at room temperature. The paper-based biochemical reagent storage method based on the water two-phase system of the present invention uses the water two-phase system as a reagent storage medium, fully utilizes its characteristics of good biocompatibility and controllable phase separation, and realizes stable storage of biochemical reagents.

Description

technical field [0001] The invention relates to the field of point-in-time testing (POCT), in particular to a storage method for paper-based biochemical reagents based on a water two-phase system, which is suitable for portable micro devices and on-site instant detection technology. Background technique [0002] Portable point-of-care detection technology is of great significance to developing countries, especially countries lacking infrastructure and medical personnel, and is also a hot spot in the fields of analytical chemistry and medicine. [0003] Paper-based microfluidic chip is the most potential technology in the field of real-time detection and analysis. It uses the capillary effect of paper fibers to drive the fluid to move along a specific line to achieve analysis and detection functions. It has the advantages of good reagent consumption, low cost, light and easy to carry, and simple operation. And other characteristics, has been applied to environmental testing, ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01L3/00
CPCB01L3/50B01L3/5027
Inventor 盖宏伟张清泉陈佳瞿
Owner HANGZHOU TINKER BIOTECHNOLOGY CO LTD
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