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Protein imprinted material and use thereof for removing albumin from human blood serum

A technology of western blotting and human serum, applied in the application field of albumin, can solve the problems of high preparation cost, dye ligand shedding, sample contamination, etc., and achieve the effect of simple operation process, reduced loss, and low production cost

Active Publication Date: 2011-04-06
中科榆林能源技术运营有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods have the following inherent defects: (1) Most of the affinity ligands are high-purity monoclonal antibodies or other molecules of biological origin, such as lectins, coenzymes, etc., and their production and purification processes are complicated, and the preparation cost is very high
(2) The physical and chemical properties of bioaffinity ligands are greatly affected by the environment, easily lose their activity, and have strict requirements on transportation, storage and operating conditions
(3) The dye ligand is easy to fall off from the matrix and cause sample contamination

Method used

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  • Protein imprinted material and use thereof for removing albumin from human blood serum
  • Protein imprinted material and use thereof for removing albumin from human blood serum
  • Protein imprinted material and use thereof for removing albumin from human blood serum

Examples

Experimental program
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Embodiment Construction

[0022] A western blot material, which can be prepared as follows:

[0023] (1) Mix functional monomers, cross-linking agents, porogens, surfactants and template proteins in phosphate buffer at pH 5-6;

[0024] (2) After the above-mentioned mixture is mixed uniformly, nitrogen gas is passed into it, and then an initiator and a speed-accelerator are respectively added therein to carry out a polymerization reaction, the polymerization temperature is controlled between 20-30° C., and the reaction time is 5-24 hours;

[0025] The amount of each substance used in the process of synthesizing imprinted materials is shown in the table below.

[0026]

[0027]

[0028] (3) After the imprinting material was synthesized according to the amount in the above table, the template protein on the polymer was eluted with a volume concentration of 9-11% acetic acid mixture containing 9-11% sodium dodecylsulfonate by weight. Subsequently, the polymer is washed with water and pH 5-6 phosphat...

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Abstract

The invention relates to a preparation method of a pig serum albumin molecularly imprinted material and application thereof to selectively removing the albumin from human serum. The method comprises the following steps: the pig serum albumin is taken as a template; a protein imprinted material is obtained by an in situ polyreaction of a functional monomer, a cross-linking agent, a pore-forming agent, a surfactant and template protein, and the characteristic that the human serum albumin has high homology with the pig serum albumin is employed to successfully applying the material to removing the albumin from the human serum. The method can help solve the problem of difficult albumin sources by taking human albumin as imprinted templates, and realize the selective removal of the albumin from the human serum by the prepared molecularly imprinted material. The invention provides a new high-abundance protein removal method for proteomics studies.

Description

technical field [0001] The invention relates to a method for removing high-abundance proteins in human serum, in particular to a western blot material and its application in removing high-abundance albumin in human serum. Background technique [0002] With the completion of the Human Genome Project, exploring the biological significance of the genome and elucidating its functions has become a research hotspot in life sciences. Proteomics (proteomics) research has a pivotal strategic position in this frontier field and has received more and more attention. In October 2001, the International Human Proteome Organization (HUPO) was established in the United States and planned to launch the Human Proteome Project (HPP). HPP is another major international scientific research project after the Human Genome Project. Its first implementation plans include: Human Liver Proteome Project (HLPP) and Human Plasma Proteome Project (Human Plasma Proteome Project, HUPOPPP) ) (Omenn, G.S., ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08F2/44C08F20/56C08J9/26
Inventor 张丽华邓启良刘晋湘马俊峰梁振张玉奎
Owner 中科榆林能源技术运营有限责任公司