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Preparation method of chiral stationary phase based on photocatalytic cross-linked protein, chiral stationary phase and application thereof

A chiral stationary phase and cross-linked protein technology, applied in the field of chiral stationary phase, can solve problems such as weak interaction and stability, and achieve the goals of increasing area, improving structural inhomogeneity, good uniformity and stability Effect

Active Publication Date: 2021-02-23
泰州学院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, its disadvantage is that CSP prepared by physical adsorption may have stability problems due to the weak interaction between the adsorbed protein and the carrier.

Method used

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  • Preparation method of chiral stationary phase based on photocatalytic cross-linked protein, chiral stationary phase and application thereof
  • Preparation method of chiral stationary phase based on photocatalytic cross-linked protein, chiral stationary phase and application thereof
  • Preparation method of chiral stationary phase based on photocatalytic cross-linked protein, chiral stationary phase and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035]For the photocatalytic cross-linking bovine serum albumin test, take a 50mL centrifuge tube, add 10mg / mL bovine serum albumin solution to it, and then add 0.0125M tris(2,2'-bipyridyl)ruthenium dichloride ( II) Hexahydrate and 0.25M ammonium persulfate (APS), and then placed under a 200W incandescent lamp for photocatalytic crosslinking reaction. And carry out the following test.

[0036] SDS-PAGE test

[0037] Refer to the "Molecular Cloning Experiment Guide" to prepare electrophoresis reagents, separating gel, and stacking gel. First assemble the glass plate and make sure to make a 1mm-thick 10-hole comb gel, and add reagents in sequence according to the formula of the separating gel in Table 1 and mix well. Pour the mixed acrylamide solution into the gap of the glass plate of the electrophoresis device, carefully add deionized water to cover the acrylamide solution, and wait for the polymerization to complete. After polymerization, rinse the unpolymerized acrylamide...

Embodiment 2

[0046] Preparation of chiral stationary phase based on photocatalytic cross-linked protein:

[0047] (1) Preparation of activated silica gel

[0048] Weigh 20g of silica gel, dissolve in 60ml of HCl (6.0M), stir and reflux at 150°C for 8h, filter the activated silica gel with suction, and wash with distilled water until neutral. Finally, the product was dried at 70°C for 8 hours, and put into a reagent bottle for later use.

[0049] (2) Sulfhydryl modification:

[0050] First, add 1.2mL of γ-mercaptopropyltrimethoxysilane into a 20mL beaker containing 10mL of 95% ethanol, mix thoroughly with a vortex mixer, adjust the pH value of the system to 5 with concentrated hydrochloric acid, and pour it into a cone shaped bottle. 15 g of activated silica gel was added, and then the Erlenmeyer flask was placed in a constant temperature shaker at 37° C., 150 rpm, and reacted for 4 hours. After taking it out, first use ethanol to clean repeatedly with ultrasonic vibration, and then ful...

Embodiment 3

[0063] Preparation of chiral stationary phase based on photocatalytic cross-linked protein:

[0064] (1) Preparation of activated silica gel

[0065] Weigh 20g of silica gel, dissolve in 60ml of HCl (6.0M), stir and reflux at 150°C for 8h, filter the activated silica gel with suction, and wash with distilled water until neutral. Finally, the product was dried at 70°C for 8 hours, and put into a reagent bottle for later use.

[0066] (2) Sulfhydryl modification:

[0067] First, add 1.2mL of γ-mercaptopropyltrimethoxysilane into a 20mL beaker containing 10mL of 95% ethanol, mix thoroughly with a vortex mixer, adjust the pH value of the system to 5 with concentrated hydrochloric acid, and pour it into a cone shaped bottle. 15 g of activated silica gel was added, and then the Erlenmeyer flask was placed in a constant temperature shaker at 37° C., 150 rpm, and reacted for 4 hours. After taking it out, first use ethanol to clean repeatedly with ultrasonic vibration, and then ful...

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Abstract

The invention discloses a preparation method of a chiral stationary phase based on a photocatalytic cross-linked protein, the chiral stationary phase and an application thereof; the preparation methodof the chiral stationary phase based on photocatalytic cross-linked protein comprises the steps: photocatalytic cross-linking: sequentially adding tris(2,2'-dipyridyl)ruthenium (II) dichloride hexahydrate and ammonium persulfate into a protein solution, and carrying out photocatalytic cross-linking reaction under illumination; immobilizing: adding the modified and coupled silica gel into a protein solution subjected to photocatalytic cross-linking, fully stirring, carrying out suction filtration, and drying, so as to obtain the chiral stationary phase. The BSA-CSP disclosed by the invention is a chiral separation stationary phase with a brand-new structure, and can be suitable for chiral resolution of chemicals including amino acid and having certain binding capacity with a protein sequence.

Description

technical field [0001] The invention belongs to the technical field of chiral separation, and in particular relates to a preparation method, chiral stationary phase and application of a chiral stationary phase based on photocatalytic cross-linked protein. Background technique [0002] Chirality, expressed as the difference between an object and its mirror image, is a special form of symmetry that exists widely in nature. Chiral objects and their mirror images are called enantiomers. The spatial arrangement order of these enantiomers is different, which often leads to differences in their physiological activities. Enantiomers with different configurations also have different pharmacological activities and side effects. showed a large difference. [0003] The content of single enantiomer is limited in nature. When this substance is needed in large quantities in life, it often needs to be synthesized artificially. However, the asymmetric synthesis of a single enantiomer is re...

Claims

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Application Information

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IPC IPC(8): B01J20/29B01J20/28B01J20/30C07B57/00C07C227/34C07C229/36C07C229/22C07C229/26C07C229/08C07D209/20C07C319/28C07C323/58C07C277/08C07C279/14
CPCB01J20/28011B01J20/29C07B57/00C07B2200/07C07C227/34C07C277/08C07C319/28C07D209/20C07C229/36C07C229/22C07C229/26C07C229/08C07C323/58C07C279/14
Inventor 张鑫冯卫伟高新星何清明彭加平韦平和
Owner 泰州学院
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