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

SCX/HIC (Strong Cation Exchange/Hydrophobic) mixed-mode chromatograph stationary phase and preparation method thereof

A mixed-mode chromatography and stationary phase technology is applied to a hydrophobic/strong cation-exchange dual-functional mixed-mode chromatography stationary phase. In the field of preparation of the mixed-mode stationary phase, problems such as limited types can be solved, and the synthesis cost is low and the service life is long. , good separation effect

Inactive Publication Date: 2012-07-04
NORTHWEST UNIV
View PDF3 Cites 12 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are still few dual-function chromatographic packing materials specially designed for separating proteins, and the types are very limited

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
  • SCX/HIC (Strong Cation Exchange/Hydrophobic) mixed-mode chromatograph stationary phase and preparation method thereof
  • SCX/HIC (Strong Cation Exchange/Hydrophobic) mixed-mode chromatograph stationary phase and preparation method thereof
  • SCX/HIC (Strong Cation Exchange/Hydrophobic) mixed-mode chromatograph stationary phase and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] （1） Take 3.3g (about 13.7 mmol) of cystine and dissolve it in 150 mL of 0.5 mol / L Na 2 CO 3 buffer solution, then adjust the pH to 11.0, add it to a 250 mL three-necked flask, and slowly add 3 mL (about 13.7 mmol) of TM560 dropwise under ice-bath stirring. 65°C, reacted for 24 hours, the reaction was completed, the solution was light yellow, cooled to room temperature, adjusted the pH of the reaction solution to 5.5 with glacial acetic acid, added 2 g of silica gel, reacted at 90°C for 2 hours, filtered, and washed three times with water, methanol, Wash with acetone and methanol, and dry the obtained solid under vacuum at 50° C. for 8 hours to obtain a cystine-bonded silica gel derivative. Take 5.0 mL of ninhydrin ethanol solution, add a small amount of cystine silica gel derivative, heat at 90°C for 5 min, and find that the solution turns purple, indicating that cystine has been successfully bonded to the silica gel surface.

[0047] (2) Disperse 2.0 g of cystine-bo...

Embodiment 2

[0055] The chromatographic filler prepared in Example 1 was used to pack the column, and then the five standard proteins were separated under the strong cation exchange mode. Separation conditions:

[0056] Mobile phase: Liquid A: 20 mmol / L KH 2 PO 4 (pH 6.5); B liquid: 20 mmol / L KH 2 PO 4 + 1.0 mol / L NaCl (pH 6.5), linear gradient elution, 0→100%B, 30 min; flow rate 1.0 mL / min, for five proteins including myoglobin, RNase B, RNase A, cytochrome c, and lysozyme good separation (as figure 2 1, 2, 3, 4, and 5 shown are myoglobin, RNase B, RNase A, cytochrome c, and lysozyme, respectively).

Embodiment 3

[0058] The chromatographic filler prepared in Example 1 was used to pack the column, and then the seven standard proteins were separated in the hydrophobic mode. Separation conditions:

[0059] Mobile phase: Liquid A: 20 mmol / L KH 2 PO 4 (pH 7.0) + 3.0 mol / L (NH 4 ) 2 SO 4 (pH 7.0); Solution B: 20 ​​mmol / L KH 2 PO 4 (pH 7.0), linear gradient elution, 0→100%B, 30 min, flow rate of 1.5 mL / min, achieved cytchrome c, myoglobin, RNase A, lysozyme, OVA, α-amylase, insulin and other seven proteins good separation (eg image 3 1, 2, 3, 4, 5, 6, and 7 shown are cytchrome c, myoglobin, RNase A, lysozyme, OVA, α-amylase, insulin, respectively).

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 strong cation exchange / hydrophobic mixed-mode chromatograph stationary phase, wherein X is -OCH3 or -OCH2CH3; R is shown in the specification, wherein n is equal to 1-5, or R is shown in the specification, wherein n is equal to 1-6; or R is PEG (Polyethylene Glycol) 200-1000. A preparation method comprises the following steps of: bonding cystine on the activated silica gel surface with hydroxyl groups by a silane coupling agent, then using DTT to open cystine disulfide bonds on the silica gel surface and form sulfydryl silica gel, then using H2O2 to oxidize the sulfydryl groups into sulfonic acid groups and form silica-gel derivatives bonded with the sulfydryl groups, and finally reacting with fatty alcohol (or aromatic alcohol or PEG and the like) to obtain the hydrophobic / strong cation exchange chromatograph stationary phase. The stationary phase can realize effective separation of proteins under the hydrophobic mode and the strong cation exchange mode, and one chromatographic column filled with a double-function separating medium can replace two common strong cation exchange / hydrophobic chromatographic columns to carry out separation and purification on the proteins.

Description

technical field [0001] The present invention relates to a high-performance liquid chromatography stationary phase for protein separation, specifically a hydrophobic / strong cation exchange dual-function mixed-mode chromatography stationary phase, and the invention also relates to the preparation method and application of the mixed-mode stationary phase . Background technique [0002] With the development of biotechnology and life sciences, both the production of recombinant protein drugs and the research of proteomics depend on the efficient and rapid separation technology of proteins. The analysis of complex samples puts forward higher and higher requirements for separation science, and the development of new high-efficiency separation materials, separation modes and high-sensitivity detection methods is one of the effective ways to solve this problem. [0003] Multidimensional liquid chromatography (MDLC) is a key technology for the separation and analysis of complex sampl...

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): B01J20/286B01J20/30C07K1/18
Inventor 白泉赵开楼
Owner NORTHWEST 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