Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

A secondary antibody based on a portable blood glucose meter

A blood glucose meter and portable technology, applied in the field of medical/food testing, can solve problems such as restricting applications and detecting a single target, and achieve the effects of reducing laboratory costs, reducing use, and expanding application space

Active Publication Date: 2021-12-14
JIANGNAN UNIV
View PDF7 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above preparations can only detect a single target, which greatly limits their application

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
  • A secondary antibody based on a portable blood glucose meter
  • A secondary antibody based on a portable blood glucose meter
  • A secondary antibody based on a portable blood glucose meter

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Construction of embodiment 1 expression strain

[0047] According to the base sequence (GeneID: 854644) of Saccharomyces cerevisiae sucrase Suc2 (amino acid sequence shown in SEQ ID NO.1), primers Suc2-F and Suc2-R were designed with Primer Premier 5.0, and the Saccharomyces cerevisiae genome was used as a template to amplify increase. The amplified fragments were recovered by a gel recovery kit. The recovered Suc2 gene fragment was double-digested with restriction endonucleases Nde I and Xho I and recovered ( figure 1 Swimming lane 3), and the commercialized vector pET22b ( figure 1 Swimming lane 2) for ligation, the product after ligation was transformed into commercialized Escherichia coli expression host BL21 (DE3), the transformed Escherichia coli was screened on a plate coated with ampicillin, and positive colonies were selected for sequencing verification, and the correct clone was verified as Expression strain of free sucrase Suc2.

[0048] In addition, in ord...

Embodiment 2

[0050] Fermentation and purification of the protein of embodiment 2

[0051] The fermentation method and purification method of Suc2, ZZ domain and the fusion protein Suc2-ZZ, ZZ-Suc2 adopt the conventional method and the conditions are the same, so the fermentation method of the fusion protein Suc2-ZZ is taken as an example to illustrate the fermentation and purification method of the protein in this example .

[0052] Take 20 μL of the fusion protein Suc2-ZZ glycerol bacteria to 3 mL of ampicillin-resistant LB medium (peptone 10 g / L, yeast powder 5 g / L, sodium chloride 10 g / L), culture overnight at 37 ° C, 200 rpm, and follow the steps of 4 % inoculum size was transferred to TB fermentation medium (peptone 12g / L, yeast powder 24g / L, glycerol 4g / L, KH 2 PO 4 23.1g / L, K 2 HPO 4 125.4g / L), until the bacterial concentration reaches OD 600 After the temperature is 0.6-1.2, the inducer IPTG with a final concentration of 0.4mM is added for induction, and the conditions for p...

Embodiment 3

[0055] Example 3 Evaluation of fusion protein sucrose hydrolysis ability

[0056] The enzyme reaction system is: 0.1M disodium hydrogen phosphate-citric acid buffer solution, pH 5.0, the final concentration of substrate sucrose is 1.5%, and the final concentration of pure enzyme is 10nM. The enzyme reaction conditions are: 40°C water bath for 5 minutes. After the reaction is complete, take 400 μL of the reaction solution and add it to a centrifuge tube pre-installed with 300 μL of DNS reagent, and place it in a boiling water bath for 5 minutes. After completion, immediately cool the test tube in ice water, and measure the absorbance with a spectrophotometer at a wavelength of 520 nm after appropriate dilution. The treatment method of the corresponding blank group is to first take 400 μL of enzyme-free reaction solution and add it to a centrifuge tube pre-installed with 300 μL of DNS reagent, and put it in a boiling water bath for 5 minutes. After appropriate dilution, the abs...

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 discloses a secondary antibody based on a portable blood glucose meter, which has two functional domains, respectively performing the functions of binding IgG antibodies and decomposing sucrose; wherein, the glucose generated after the sucrose is decomposed can be detected by the portable blood glucose meter. The present invention successfully realizes the soluble expression of the secondary antibody in Escherichia coli, which proves that the secondary antibody is not inferior to free sucrase and ZZ domain in terms of sucrose hydrolysis ability and IgG antibody binding ability. Moreover, the present invention successfully uses the secondary antibody to detect relevant antigen indicators, and the detection results are in line with theoretical results. It laid the foundation for its subsequent application in point-of-care testing technology.

Description

technical field [0001] The invention belongs to the technical field of medicine / food detection, and specifically relates to a secondary antibody based on a portable blood glucose meter. Background technique [0002] In clinical testing and research on life science-related issues, various immunoassay techniques such as the most common enzyme-linked immunoassay (ELISA) are often involved. In ELISA, due to the different sources of primary antibodies, different secondary antibodies need to be matched, which directly leads to an increase in the cost of related experiments. In addition, the secondary antibodies currently on the market are either modified by alkaline phosphatase, or modified by fluorescent groups, or modified by peroxidase, all of which require large-scale equipment to characterize the results, not only limited by space, but also And it requires the tester to have certain professional knowledge. In order to develop a simple and low-cost analytical method for the ...

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
Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/42C12N9/24C07K19/00C12N15/70G01N33/68G01N33/66
CPCC07K16/42C12N9/2405C12Y302/01048C12N15/70G01N33/6854G01N33/66C07K2319/00
Inventor 吴志猛周志昉洪皓飞董笑飞鲍奕恺胡苗苗叶可嘉李舜岩俞杭艳
Owner JIANGNAN UNIV
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com