Method for rapidly screening ERp57 inhibitors by high throughput

An inhibitor, high-throughput technology, applied in the field of biomedicine, can solve the problems of high cost, low screening efficiency, environmental pollution, etc., and achieve the effect of low cost and sample saving.

Inactive Publication Date: 2018-11-27
ZUNYI MEDICAL UNIVERSITY
View PDF8 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Such a method takes a long time, has low screening efficiency, heavy workload, and high cost; subsequently developed gelatin zymography, radioisotope method, high performance liquid chromatography (HPLC) assay, spectrophotometry, etc. There are corresponding defects, such as the radioisotope method takes a long time and pollutes the environment; HPLC assay requires a large amount of mobile phase and expensive chromatographic columns, etc.

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
  • Method for rapidly screening ERp57 inhibitors by high throughput
  • Method for rapidly screening ERp57 inhibitors by high throughput
  • Method for rapidly screening ERp57 inhibitors by high throughput

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0020] The following examples further illustrate the content of the present invention, but should not be construed as limiting the present invention. Any modifications or substitutions made to the methods, steps or conditions of the present invention without departing from the essence of the present invention fall within the scope of the present invention. Unless otherwise specified, the technical means used in the embodiments are conventional means well known to those skilled in the art.

[0021] All reagents used in the examples are commercially available unless otherwise specified.

[0022] The sample to be screened can be a traditional Chinese medicine extract or a mixture of multiple compounds. In this patent, the extract of the traditional Chinese medicine Salvia miltiorrhiza is taken as an example.

[0023] 1. Preparation of ERp57 protein with His tag:

[0024] Add 10 ml of binding buffer to suspend 2 g wet weight of E. coli BL 21 strain (transformed with the constru...

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 relates to the field of biological medicines, and particularly discloses a high-throughput drug screening method based on a combination principle. The high-throughput drug screening method based on the combination principle comprises the following steps: (1) preparing target proteins with histidine-tags; (2) incubating to-be-screened samples with protein; (3) combining the mixture with agarose magnetic beads chelated with nickel; (4) magnetically separating the magnetic beads, eluting non-specific conjugates on the surfaces of the magnetic beads; (5) eluting a target compound, and identifying micromolecules combined to the target protein by using a mass spectrograph; and (6) further verifying biological activity of the micromolecules which are combined to the target protein.If a to-be-measured sample has a biological regulation function of the protein, then the to-be-measured sample is positive. The method has the characteristics of rapidness, low cost, capability of saving samples and the like, and a new screening method for discovering a target compound is provided.

Description

technical field [0001] The invention relates to the field of biomedicine, and relates to a high-throughput screening method for protein ligands based on the principle of nickel ion chelated agarose magnetic beads affinity chromatography. Background of the invention [0002] The traditional method for drug screening of small molecule inhibitors is to evaluate the biological activity of the tested compounds one by one. Such a method takes a long time, has low screening efficiency, heavy workload, and high cost; subsequently developed gelatin zymography, radioisotope method, high performance liquid chromatography (HPLC) assay, spectrophotometry, etc. There are corresponding defects, such as the radioisotope method takes a long time and pollutes the environment; the HPLC assay requires a large amount of mobile phase and expensive chromatographic columns. Therefore, the development of fast, effective, simple and high-throughput screening methods is an inevitable trend to realize...

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 Applications(China)
IPC IPC(8): G01N27/62G01N30/02
CPCG01N27/62G01N30/02
Inventor 崔国祯周鹤峰师伟邹佳毛皓李惠民
Owner ZUNYI MEDICAL UNIVERSITY
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