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Paper-based biochemical reagent storage method based on water two-phase system

A two-phase system and biochemical reagent technology, applied in chemical instruments and methods, laboratory containers, analytical material containers, etc., can solve the problem of inconvenient storage of reagents, achieve a wide range of applications, achieve stable storage, and be beneficial to business The effect of promotion

Active Publication Date: 2020-09-18
HANGZHOU TINKER BIOTECHNOLOGY CO LTD
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  • Description
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AI Technical Summary

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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  • Paper-based biochemical reagent storage method based on water two-phase system
  • Paper-based biochemical reagent storage method based on water two-phase system
  • Paper-based biochemical reagent storage method based on water two-phase system

Examples

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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 comprises the steps that a biochemical reagent to be stored is dissolved in a water two-phase system solution, then the water two-phase system solution is dropwise added into a reagent storage area of a paper chip, solvent volatilization induces the water two-phase system to be subjected to phase separation, and a water-containing double-layer film is formed. The biochemical reagent is selectively distributed on the lower-layer film, the upper-layer film inhibits moisture volatilization of the lower-layer film,direct contact between the reagent and the external environment is avoided, and the storage stability of the biochemical reagent is improved. Then, the paper chip loaded with the reagent is sealed ina vacuum sealing bag, and can be stored at 4 DEG C or normal temperature. According to the paper-based biochemical reagent storage method based on a water two-phase system, the water two-phase systemserves as a reagent storage medium, the characteristics of good biocompatibility and controllable phase separation of the water two-phase system are fully utilized, and stable storage of biochemical reagents is achieved.

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 Applications(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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