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Method for synchronous enriching and desalinating bovine serum albumins by using novel solid phase extractant

A technology of bovine serum albumin and solid-phase extraction agent, applied in the field of biochemical analysis and identification, can solve the problems of dialysis and electrodialysis, which are time-consuming, difficult to scale up large-scale production, and impossible to achieve separation, etc., to achieve easy industrial production, Low-cost and time-consuming effects

Inactive Publication Date: 2016-11-09
SUZHOU BAIRUI BIOLOGICAL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Dialysis and electrodialysis take a long time, the sample dilution is large, and it is not easy to scale up for large-scale production; gel filtration has certain limitations, such as when the molecular weight of salt and protein is similar, it is impossible to achieve the separation effect

Method used

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  • Method for synchronous enriching and desalinating bovine serum albumins by using novel solid phase extractant
  • Method for synchronous enriching and desalinating bovine serum albumins by using novel solid phase extractant
  • Method for synchronous enriching and desalinating bovine serum albumins by using novel solid phase extractant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] A kind of preparation method of graphene microsphere solid-phase extraction agent, concrete steps are:

[0023] Step 1: Add deionized water to graphene oxide, prepare a graphene oxide solution with a mass concentration of 3.0wt%, stir it under a magnetic stirrer for 8 hours, and then ultrasonicate it in an ultrasonic machine for 3 hours to obtain dispersed graphene oxide solution;

[0024] Step 2: Put the mixed solution in an ultrasonic nebulizer connected with a quartz tube, and the ultrasonic frequency is 1.5MHz. Make it atomized into aerosol droplets, in N 2 Driven by a vacuum pump and a suction filter, it slowly passes through a quartz tube heated to 550°C, and the end is collected with a PTFE filter membrane. Dry at room temperature for 3 hours, then put it in a vacuum desiccator and dry at 60°C for 12 hours to obtain a graphene microsphere solid-phase extraction agent with a particle size of 1 μm-2 μm, such as figure 1 shown.

Embodiment 2

[0026] Carry out compression resistance experiment with the graphene microsphere gained in embodiment 1, concrete steps are:

[0027] Three kinds of adsorbents were selected as graphene microspheres, C18 silicon and carbon nanotubes, four pressures of 10 MPa, 20 MPa, 30 MPa and 40 MPa were set, and the pressure action time was 3 minutes. A pressure test is carried out on a material, and the material under pressure is immediately observed under a scanning electron microscope. It was found that the graphene microspheres still maintained their original appearance under the four pressure environments, and their structure did not change, indicating that the compression resistance was strong; C18 silicon had a small amount of fragmentation under the condition of 10 MPa, and as the pressure increased, the fragmentation The degree of carbon nanotubes gradually intensifies. When the pressure is 40 MPa, they are all in powder form, and the compression resistance is poor. Flaky, loses i...

Embodiment 3

[0029] Using the graphene microspheres obtained in Example 1 as a solid-phase extractant for enrichment and desalination of bovine serum albumin, the specific steps are:

[0030] Step 1: Weigh 20 mg of graphene microspheres and add them to a solid-phase extraction column with a volume of 3 ml, seal the upper and lower sieve plates, and activate the column with 10 ml of methanol solution with a volume percentage of 70%, and control the flow rate to 1.2ml / min;

[0031] Step 2: Add sodium chloride with a mass ratio of 5% to the bovine serum albumin solution, use a constant-flow pump to pass the salt-containing protein solution through the solid-phase extraction column, control the flow rate to 1.2ml / min, and add 3ml volume after loading the sample Rinse with 10% methanol solution to remove unadsorbed protein, and vacuum to dryness. Use 2ml of acetonitrile as the protein eluent, control the flow rate to 0.2ml / min, and collect the eluate. Select non-solid-phase extraction soluti...

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Abstract

The invention relates to a method for synchronous enriching and desalinating bovine serum albumins by using a novel solid phase extractant. The method comprises the following steps: carrying out ultrasonic atomization on a graphene oxide dispersion, allowing the atomized graphene oxide dispersion to go through a 450-550DEG C high temperature furnace, drying the obtained atomized graphene oxide dispersion, and collecting the dried atomized graphene oxide dispersion to obtain a graphene sphere solid phase extractant; adding solid sodium chloride powder to a bovine serum albumin solution, allowing the salt-containing protein solution to go through a solid phase extraction column, leaching the salt-containing protein solution, carrying out vacuum pumping until the column is dry, eluting the dried salt-containing protein solution, and collecting the obtained eluate; and carrying out micro-porous filtration on the eluate, and carrying out AKTA-Start analysis detection. The graphene microsphere solid phase extractant prepared in the invention has the advantages of simple technology, low cost, large specific surface area, realization of convenient, fast and high-dosage enrichment of proteins, realization of synchronous implementation of enrichment and desalination, provision of an economic and rapid new method for desalination of the proteins, and great application potential, and has an obviously better anti-pressure ability than commercial adsorbents.

Description

technical field [0001] The invention relates to the technical field of biochemical analysis and identification, in particular to a method for synchronous enrichment and desalination of bovine serum albumin by a novel solid-phase extraction agent. Using graphene microspheres as an adsorbent can enrich proteins conveniently, quickly, and in high doses, achieving simultaneous enrichment and desalination. Background technique [0002] Graphene, a new class of carbon nanomaterials, has attracted great interest from scientists due to its unique two-dimensional nanostructure. Graphene has extraordinary electrical, thermodynamic and mechanical properties, such as ultra-high specific surface area, good thermal conductivity, ultra-fast charge mobility, and Young's modulus, which make it of great application value in chemical analysis. Wrinkled graphene balls still have a π-conjugated structure, so they can have a stronger affinity with polycyclic aromatic hydrocarbons. More important...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/40
CPCG01N1/405
Inventor 聂华丽刘琳
Owner SUZHOU BAIRUI BIOLOGICAL SCI & TECH CO LTD
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