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Preparation method of carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) magnetic protein grains and application thereof

A magnetic particle and protein technology, applied in the fields of molecular biology and applied microbiology, can solve the problems of difficult regeneration of activated carbon, high processing cost, and small processing volume, and achieve the effects of efficient method, simple preparation method, and low production cost

Inactive Publication Date: 2013-04-24
FIRST AFFILIATED HOSPITAL OF KUNMING MEDICAL UNIV
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Problems solved by technology

Among them, the mechanical removal method has good algae removal effect and large treatment capacity, but the cost is high; manual salvage, ultrasonic and electromagnetic wave algae removal have small treatment capacity, and are only suitable for small-scale algae removal; activated carbon adsorption has a good effect on algae removal, but activated carbon regeneration Difficult and, therefore, expensive to process

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  • Preparation method of carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) magnetic protein grains and application thereof
  • Preparation method of carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) magnetic protein grains and application thereof
  • Preparation method of carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) magnetic protein grains and application thereof

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Embodiment Construction

[0020] Using the human hepatocyte cDNA library as a template, the target gene human CEACAM1 (NCBI AccessionNo.: NM_001712) was amplified by PCR, and the primer sequences used were:

[0021] Ceacam1-F:5'-GAAA GGATCC ATGAGCTGGTACAAAGGGGA-3',

[0022] Ceacam1-R:5'-GAAA CTCGAG TTAGGTGTCTGAAGGGGAAATG-3'.

[0023] The human CEACAM1 coding region sequence treated with BamHI / XhoI and pET-28a(+) were ligated to construct an expression vector. The constructed expression vector was transformed into E.coli BL21(DE3), and after being induced by IPTG, E.coli BL21(DE3) massively expressed human CEACAM1 in the form of inclusion body. After the inclusion bodies were washed and resuspended, human CEACAM1 with higher purity was obtained through denaturation, gel filtration chromatography, refolding and secondary gel filtration chromatography in sequence. with Fe 3 o 4 As a carrier, after being coated with silicic acid, it reacts with silane in a high-temperature acidic environment and r...

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Abstract

The invention relates to an expressing and purifying method and application of protein fused with an extracellular special structure domain of a carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1). The preparation method disclosed by the invention comprises the following steps: adopting the hepatocyta cDNA library as a template to amplify the target human genome CEACAM 1; adopting the heterologous expression technique to express CEACAM 1 protein fused with the special structure domain outside the cells in escherichia coli (E.coliBL21); carrying out the resilience and gel filtration chromatography for the protein so as to obtain soluble CEACAM 1 fusion protein; and adopting the Fe3O4 as a carrier, coating with silicic acid, reacting with silane in an acid environment, and reacting with glutaric anhydride in an alkaline environment, thus finally obtaining the CEACAM1 protein magnetic grains. A lot of extracellular special structure domains of the carcinoembryonic antigen-related cell adhesion molecules 1 (CEACAM1) with high purity can be produced by the method; and the micro / nano magnetic particle-CEACAM 1 compound is prepared from the special structure domains as basic ingredients. Research at the present stage shows that the compound can be used for quickly separating and detecting Neisseria gonorrhoeae and blue algae, so that the compound has good application prospect.

Description

technical field [0001] The invention relates to a method for expressing, purifying and applying a human carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) extracellular specific domain fusion protein, belonging to the fields of molecular biology and applied microbiology. Background technique [0002] Carcinoembryonic antigen-associated cell adhesion molecule 1 (CEACAM1, NCBI GenBank: NM_001712) is a transmembrane glycoprotein widely expressed on the surface of neutrophils, macrophages, epithelial cells and lymphocytes, and belongs to the carcinoembryonic antigen family immunoglobulin Superfamily adhesion molecules, whose biological functions include promoting the formation of blood vessels, regulating the remodeling of blood vessels, participating in the regulation of apoptosis, promoting the formation of gland lumen and regulating the clearance of insulin to maintain the sensitivity of cells to insulin, At the same time, CEACAM1 is also an adhesion receptor...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/70C07K17/14C12Q1/02C12R1/36C12R1/89
Inventor 孟照辉谢月辉李玉叶叶秋芳
Owner FIRST AFFILIATED HOSPITAL OF KUNMING MEDICAL UNIV
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