Production of metal alloyed nano-zeolite affinity chromatography separating material

A nano-zeolite and metal doping technology, which is applied in the field of nanotechnology preparation, can solve the problems of affecting the biological stability of biological macromolecules, the loss of affinity ligands such as metal ions, and the difficulty of increasing the leaching speed, etc., to achieve excellent physical properties. Chemical stability, avoiding degradation and structural interference changes, low back pressure effect

Inactive Publication Date: 2010-04-14
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Most of the traditional immobilized metal ion affinity chromatography technology uses natural or synthetic polymer chromatography matrix materials such as dextran gel and agarose gel, which obviously have poor mechanical strength and it is difficult to improve the elution speed; it is easy to breed dangerous microorganisms shortcoming
[0003] Find through the document retrieval to prior art, J.G.Krabbe etc. published " Selective detection and identification of phosphorylated proteins by Simultaneousligand-exchange fluorescence detection and mass spectrometry" ("Simultaneous ligand-exchange fluorescence detection and mass spectrometry method for selective analysis and identification of phosphorylated proteins"), reported the use of polymer chromatography by immobilized metal ion affinity chromatography The matrix material separates phosphorylated protein molecules, but since the affinity ligands such as metal ions are combined with the corresponding matrix material through chelation between ions, there will be loss of affinity ligands such as metal ions during the washing process, etc. Bottleneck” problem will directly affect the biological stability of biological macromolecules such as target protein molecules in the biological separation system, so that its further practical application is limited

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] (1) Weighing solid FeCl 3 .6H 2 O 0.196g, dissolved in 5.12g deionized water, to obtain a concentration of 0.14moL / L yellow FeCl 3 The solution is ready for use;

[0016] (2) Weigh 0.2934g of solid tetrabutyl titanate, add 12.373g of 25% tetraethylammonium hydroxide dropwise (the molar ratio of the two is 0.06:1), and the solution is 12.7mL in total, and then fully stirred by a magnetic stirrer 3 hours, until the solution is transparent;

[0017] (3) Add 2.346g of white carbon black with a mass percentage of 99% in the solution obtained in step (2), stir rapidly to obtain a white suspension solution, and the above-mentioned prepared FeCl 3 solution was added dropwise to this solution, so that FeCl 3 The final concentration of the solution is 0.04moL / L, and it is stirred rapidly and fully;

[0018] (4) Completely transfer the solution obtained in step (3) to a stainless steel reaction kettle lined with a polytetrafluoroethylene reaction vessel, and conduct a hydroth...

Embodiment 2

[0022] (1) Weighing solid FeCl 3 .6H 2 O 0.196g, dissolved in 6.53g deionized water, to obtain a concentration of 0.11moL / L yellow FeCl 3 The solution is ready for use;

[0023] (2) Weigh 0.2 g of solid tetrabutyl titanate, add 20 g of 15% tetraethylammonium hydroxide dropwise (the molar ratio of the two is 0.04:1), the solution is 20 mL in total, and then fully stir for 3.5 hours by a magnetic stirrer , until the solution is transparent;

[0024] (3) Add 2.2g of white carbon black with a mass percentage of 90% in the solution obtained in step (2), stir rapidly to obtain a white suspension solution, and the above-mentioned prepared FeCl 3 solution was added dropwise to this solution, so that FeCl 3 The final concentration is 0.03moL / L, and stir rapidly;

[0025] (4) Completely transfer the solution obtained in step (3) to a stainless steel reaction kettle lined with a polytetrafluoroethylene reaction vessel, and conduct a hydrothermal crystallization reaction at a tempera...

Embodiment 3

[0028] (1) Weighing solid FeCl3 .6H 2 O 0.196g, dissolved in 5.12g deionized water, to obtain a concentration of 0.11moL / L yellow FeCl 3 The solution is ready for use;

[0029] (2) Weigh 0.3 g of solid tetrabutyl titanate, add 13 g of 25% tetraethylammonium hydroxide dropwise (the molar ratio of the two is 0.055:1), the solution is 13 mL in total, and then fully stir for 4 hours by a magnetic stirrer , until the solution is transparent;

[0030] (3) Add 3.7g of white carbon black with a mass percentage of 80% in the solution obtained in step (2), stir rapidly to obtain a white suspension solution, and the above-mentioned prepared FeCl 3 solution was added dropwise to this solution, so that FeCl 3 The final concentration is 0.035moL / L, and stir rapidly;

[0031] (4) The solution obtained in step (3) is completely transferred to a stainless steel reaction kettle lined with a polytetrafluoroethylene reaction vessel, and then subjected to a hydrothermal crystallization reactio...

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Abstract

A metal doped nano-zeolite affinity chromatographic separation material is prepared through proportionally dissolving FeCl3.6H2O in deionized water to obtain FeCl3 solution, adding ammonium tetraethylhydroxide to tetrabutyl titanate, stirring until the solution becomes transparent, adding white carbon black, stirring to obtain white suspension, adding said FeCl3 solution, stirring, hydrothermal crystallizing in oven, cyclically washing the resultant with deionized water, and drying. It can be used for specifically enriching and separating phosphopeptide and phsphoprotein.

Description

technical field [0001] The invention relates to a preparation method in the field of nanotechnology, in particular to a preparation method of a metal-doped nano zeolite affinity chromatography separation material. Background technique [0002] With the continuous deepening of modern life science research, the role and significance of functional biomacromolecules in complex biological sample systems in life activities are clearly clarified, so as to further explore the causes of major diseases that seriously threaten human health, such as cancer and cardiovascular diseases. The pathogenic mechanism has become a hot issue concerned by many related disciplines such as life sciences, medicine, and pharmacology. In this research process, the establishment of an effective separation and purification technology for functional protein molecules is a very critical link. The reversible process of protein phosphorylation and dephosphorylation is involved in almost all life activities ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): B01J20/283B01J20/18
Inventor 徐芳王德举杨芃原张蔚霞杨明
Owner SHANGHAI JIAO TONG UNIV
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