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

A method for analyzing the binding rate of recombinant human insulin and its analogs or conjugates to insulin receptors

A technology for recombining human insulin and insulin receptors, which is applied in analytical materials, biomaterial analysis, instruments, etc., can solve the problems of cumbersome steps and achieve the effects of simplified experimental steps, good safety, and no radioactive pollution

Active Publication Date: 2021-09-21
CHONGQING PEG BIO BIOTECH CO LTD
View PDF8 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This method is practical, has high accuracy, and has no radioactive hazards, and solves the problem of radiation and cumbersome steps in the existing methods

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 method for analyzing the binding rate of recombinant human insulin and its analogs or conjugates to insulin receptors
  • A method for analyzing the binding rate of recombinant human insulin and its analogs or conjugates to insulin receptors
  • A method for analyzing the binding rate of recombinant human insulin and its analogs or conjugates to insulin receptors

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Embodiment 1 Enzyme-linked immunoassay (ELISA method) measures recombinant human insulin receptor binding rate

[0045] The recombinant human insulin receptor plasmid was constructed by Shanghai Jikai Gene Technology Co., Ltd. The HEK-293 cells used were donated by the School of Chemistry and Biological Engineering of Chongqing University of Technology. HEK-293-IR-B cells were transfected and screened by our company and purchased by Biotin From Shanghai Dongren Chemical Technology Co., Ltd., Insulin-biotin is labeled by our company. The recombinant human insulin standard (from the China Institute for Food and Drug Control, each labeled content is 20mg / tube 28.3IU / mg), recombinant human insulin is produced by Company preparation. The method for determining the binding rate of recombinant human insulin receptor comprises the following steps:

[0046] (1) After cell transfection, culture HEK-293-IR-B cells with complete medium in the logarithmic growth phase. When the cel...

Embodiment 2

[0062] Embodiment 2 Enzyme-linked immunoassay (ELISA) method measures insulin receptor binding rate and 125 Comparison of I-labeled Insulin Radiometric Determination of Binding Rate of Recombinant Human Insulin Receptor

[0063] At the same time, two methods were used to determine the binding rate of recombinant human insulin receptor, one was enzyme-linked immunoassay (ELISA), and the other was 125 I labeled insulin radiation method.

[0064] Enzyme-linked immunoassay (ELISA) method for measuring insulin receptor binding rate: Utilize the method of embodiment 1 to measure recombinant human insulin receptor binding rate (see figure 2 )

[0065] 125 I-labeled insulin radiation method to determine the binding rate of recombinant human insulin receptor:

[0066] ①After the adult Wistar rats were neck-broken to collect blood, dissect the liver immediately, remove the connective tissue, cut it into pieces, rinse it with ice-cold normal saline until there was no blood, filter i...

Embodiment 3

[0074] Example 3 Enzyme-linked immunoassay (ELISA) method to measure recombinant human insulin, insulin aspart, insulin lispro, insulin glargine, insulin detemir and PEG and modified conjugate (PEG-Lispro) receptor binding rate

[0075] Utilize the method for the enzyme-linked immunosorbent immunoassay (ELISA) of embodiment 1 to measure insulin aspart (from Novo Nordisk Company Novo sharp 300 units / 3ml / support), insulin lispro (from Eli Lilly Company Humalog 300 units) / 3ml / bottle), insulin degludec (from Novo Nordisk, trade name: Tresiba 300 units / 3ml / bottle), recombinant human insulin (sourced from China National Institutes for Food and Drug Control, each labeled content is 20mg / 28.3IU / mg), insulin glargine (sourced from Sanofi-Aventis Lantus 300 units / 3ml / branch), insulin detemir (sourced from Novo Nordisk NovoPing 300 units / 3ml / branch) ), the insulin receptor binding rate of PEG-Lispro (according to the patent EP2476430A1, self-made by the company), after 5 repeated expe...

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 method for analyzing the binding rate of human insulin and its analogs or conjugates to insulin receptors. First, the insulin receptors are transfected onto HEK-293 cells; antibodies to human insulin receptors are coated on enzyme labels On the plate, the insulin receptor antibody binds to the insulin receptor, human insulin and its analogs or conjugates compete with biotin-labeled insulin for binding to the insulin receptor, avidin binds to biotin, and the receptor binding rate is determined by HRP chromogenicity. The present invention utilizes the principles of antigen-antibody binding, ligand-receptor binding, and biotin binding to avidin to accurately measure the change in insulin receptor binding rate. No pollution, low cost, high accuracy and other requirements.

Description

technical field [0001] The present invention relates to the technical field of in vitro pharmacodynamic experimental detection of insulin receptor binding rate, in particular to the technical field of in vitro pharmacodynamic experimental detection of recombinant human insulin analogue or conjugate receptor binding rate, including fast-acting insulin such as insulin aspart (Insulin aspart), Insulin Lispro, Insulin Glulisine, long-acting insulins such as Insulin glargine, Insulin Detemir, Insulin Degludec and modified conjugates such as Method for determining the binding rate of PEGylated insulin (such as PEG-Lispro, PEG-Insulin) and PEG-Insulin (PEG conjugated des B30-insulin) to human insulin receptor. Background technique [0002] Insulin is a double-chain (A, B chain) protein hormone composed of 51 amino acids, with a molecular weight of 5374 and an isoelectric point of 5.35. The A chain contains 21 amino acids, and the B chain has 30 amino acids. The two peptide chains ...

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): G01N33/543
CPCG01N33/54306G01N2333/62
Inventor 何跃杨黎曹宇刘日勇范开
Owner CHONGQING PEG BIO BIOTECH CO LTD
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