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

Method adopting porous surface substrate to accelerate protein crystal out-phase nucleation

A technique of porous surface and heterogeneous nucleation, which is applied in peptide preparation methods, chemical instruments and methods, solution crystallization, etc., and can solve the problem of low success rate of screening

Inactive Publication Date: 2009-12-16
NORTHWESTERN POLYTECHNICAL UNIV
View PDF0 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The common problem in the screening of protein crystallization conditions is that the screening success rate is relatively low

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Example 1: Porous Surface Substrates for Lysozyme Protein Reproducibility Experiments.

[0015] Step 1: Configure hydrofluoric acid solutions with a volume ratio of 70%, 60%, 50%, and 40%, and pack them in different plastic containers. Use bamboo tweezers to soak the 18cm×18cm cover glass in the prepared solution for 10 seconds, then soak it in saturated sodium hydroxide solution for half an hour, take it out and add a little cleaning agent to ultrasonically clean it for 1 hour, and rinse it with tap water 5 times. Dry in an oven with a temperature control of 60°C for 1 day. After the silicification solution is silicified, take it out and add a small amount of cleaning agent to ultrasonically clean it for 1 hour, rinse it with tap water 10 times, and rinse it with ultrapure water 3 times. Four porous surface substrates with different surface morphologies were obtained.

[0016] Step 2: Use the Pico ScanTM 2500 atomic force microscope to scan four porous surface subst...

Embodiment 2

[0021] Example 2: Screening experiment of proteinase K crystallization conditions using porous surface substrates.

[0022] Step 1: Configure hydrofluoric acid solutions with a volume ratio of 70%, 60%, 50%, and 40%, and pack them in different plastic containers. Use bamboo tweezers to soak the 18cm×18cm cover glass in the prepared solution for 10 seconds, then soak it in saturated sodium hydroxide solution for half an hour, take it out and add a little cleaning agent to ultrasonically clean it for 1 hour, and rinse it with tap water 5 times. Dry in an oven with a temperature control of 60°C for 1 day. After the silicification solution is silicified, take it out and add a small amount of cleaning agent to ultrasonically clean it for 1 hour, rinse it with tap water 10 times, and rinse it with ultrapure water 3 times. Four porous surface substrates with different surface morphologies were obtained.

[0023] Step 2: Use the Pico ScanTM 2500 atomic force microscope to scan four...

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 accelerating protein out-phase nucleation. Hydrofluoric acid solution whose volume ratio is 70%, 60%, 50% and 40% is used to treat cover glass surface to obtain four kinds of porous surface substrate material with different features in order; protein solution performs nucleation and grows on the cover glass of the porous surface treated with the above step; the processed protein solution is put in a temperature controller with temperature of 20 DEG C for 2 days; the protein solution is taken out and observed under a microscope to calculate numbers of growing conditions of protein crystal on a porous surface substrate. Due to the adoption of porous surface as nucleation substrate, the success ratio of the protein crystal screening is improved to 46-76% from 2% of background technology.

Description

technical field [0001] The invention relates to a method for promoting heterogeneous nucleation of protein, in particular a method for promoting heterogeneous nucleation of protein crystals by adopting a porous surface substrate, which is used for repeatability research of protein crystallization and screening of crystallization conditions. Background technique [0002] The analysis of the three-dimensional structure has far-reaching significance for the study of the function of biological macromolecules. Especially as a drug target, these studies are also conducive to more rational drug design and understanding of the drug's mechanism of action. Obtaining protein crystals of high quality and suitable size is the bottleneck of structural analysis, and promoting heterogeneous nucleation of proteins is conducive to obtaining high-quality crystals. [0003] Document 1 "B.Segelke.(2001) Efficiency analysis of sampling protocols used in protein crystallization screening[J].J Cry...

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): C07K1/00B01D9/02
Inventor 尹大川郭云珠王玺凯鹿芹芹刘君
Owner NORTHWESTERN POLYTECHNICAL 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