Disulfide bond detection method and disulfide bond-containing sputum detection kit

A detection kit and detection method technology, applied in the field of biochemistry, can solve problems such as chromogenic analysis interference, lack of disulfide bonds, and rapid detection of unfavorable drug disulfide bonds, etc., achieving good specificity and repeatability , High sensitivity, avoid background interference effect

Active Publication Date: 2021-02-02
ZHEJIANG JFK BIOLOGICAL TECH
View PDF19 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Other soluble reducing agents that can reduce and break disulfide bonds, such as vitamin C (VC), tricarboxyethylphosphine (TCEP), etc., will also give potassium ferricyanide-Fe 3+ Background interference caused by chromogenic analysis of oxidative systems, which is not...

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Disulfide bond detection method and disulfide bond-containing sputum detection kit
  • Disulfide bond detection method and disulfide bond-containing sputum detection kit
  • Disulfide bond detection method and disulfide bond-containing sputum detection kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] Potassium ferricyanide concentration on the detection test paper of embodiment 1

[0085] (1) Experimental reagents and samples

[0086] Reducing agent: 2.0g reduced iron powder

[0087] Buffer solution: 100mM calcium chloride, 1.2mM MES solution, pH 6.2.

[0088] Positive sample: 2mL 0.05g / mL cystine solution; Negative sample: 2mL 0.05g / mL methionine solution

[0089] Test strip: Add 5 μL potassium ferricyanide dropwise to the result indicating area, the concentrations are 0.2 / 0.4 / 0.6 / 0.8 / 1.0 / 1.09M respectively.

[0090] (2) Sample test

[0091] Divide into 12 groups, transfer 6 groups of positive samples and 6 groups of negative samples to collection tubes, add 2mL buffer solution, and 2g iron powder. Shake the centrifuge tube for 5 minutes, let it stand for 1 minute, insert different groups of test strips vertically 0.5 cm below the liquid surface, and observe the color change of the test strip result indicator area after standing for 1-2 minutes. Color changes ...

Embodiment 2

[0097] Potassium ferricyanide dripping volume optimization on the test paper of embodiment 2

[0098] (1) Experimental reagents and samples

[0099] Reducing agent: 2.0g reduced iron powder

[0100] Buffer solution: 100mM calcium chloride, 1.2mM MES solution, pH 6.2.

[0101] Positive sample: 2mL 0.05g / mL cystine solution; Negative sample: 2mL 0.05g / mL methionine solution

[0102] Test strip: Add 1 / 3 / 5 / 7 / 10μL potassium ferricyanide dropwise to the result indicating area, the concentration is 1.09M.

[0103] (2) Sample test

[0104] Divide into 10 groups, transfer positive samples and negative samples to collection tubes, add 2mL buffer solution, and 2g iron powder. Shake the centrifuge tube for 5 minutes, let it stand for 1 minute, insert the test strips of different groups vertically 0.5 cm below the liquid surface, and observe the color change of the indicator area of ​​the test strips after standing for 1-2 minutes. Color changes are indicated with "-" and "+". "-" me...

Embodiment 3

[0110] The influence of the selection of embodiment 3 metal reducing agent on disulfide bond detection

[0111] (1) Experimental reagents and samples

[0112] The detection method of the disulfide bond comprises a reducing agent and a buffer solution.

[0113] Reducing agent: different metal reducing agents (iron powder, zinc powder, tin powder)

[0114] Buffer solution: 100mM calcium chloride, 1.2mM MES solution, pH 6.2.

[0115] Potassium ferricyanide test strip: Cut out a filter paper strip with a size (length x width) of 20mm x 5mm. About 11mm away from the bottom of the test strip, mark the potassium ferricyanide line with a dotted line, add 5 μL of saturated potassium ferricyanide solution dropwise to the area about 5mm above the dotted line and dry it, and at about 5mm above the potassium ferricyanide line, mark with Quality control indicator line with short lines. The result indication area is the area of ​​the test strip about 9-11mm from the bottom of the test st...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention provides a disulfide bond detection method and a disulfide bond-containing sputum detection kit. The disulfide bond detection method comprises the following steps of mixing a sample to be detected with a buffer system and iron powder, and then detecting disulfide bonds in the sample by developing potassium ferricyanide, wherein the buffer system is a buffer solution of divalent cations, and the pH value of the buffer system is 5.8-6.4. The method disclosed by the invention is simple and easy to implement, capable of completing detection within 10 minutes, clear and easy to judgein result, high in sensitivity and good in specificity and repeatability, and reagents used in the method are all conventional reagents and have very good safety. The kit disclosed by the invention can be stored for a long time, is convenient to carry and use, can be sealed to protect the environment after being used, and is suitable for sputum real-time collection and rapid real-time detection (POCT).

Description

technical field [0001] The invention belongs to the technical field of biochemistry, and relates to a method for detecting the reduction of disulfide bond iron powder and a sputum detection kit which are easy to use and have good safety. Background technique [0002] Disulfide bond (S-S bond) is an important covalent functional group, which exists in various natural small molecular compounds and biological protein structures. The disulfide bonds contained in these substances can usually be reductively broken to generate sulfhydryl groups (-SH), and can be interconverted with sulfhydryl groups to exist in the body fluids of the human body in a dynamic equilibrium manner, catalyzed by thiol / disulfide bond oxidoreductases. , and its content changes are often associated with specific pathological states and diseases, making them an important marker for evaluating disease risk and detecting disease states. Proteins contain disulfide bonds from cystines formed from their cysteine...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): G01N21/78G01N21/82
CPCG01N21/78G01N21/82
Inventor 陈燃徐逸丽
Owner ZHEJIANG JFK BIOLOGICAL TECH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap