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Recombinant cell with high expression of mrgprx2 and stationary phase of membrane receptor with high expression of mrgprx2 and its preparation method and application

A technology of recombinant cells and high expression, applied in the direction of carrier binding/immobilizing peptides, botany equipment and methods, biochemical equipment and methods, etc., to achieve the effect of improving sensitivity and specificity, and complete molecular activity

Active Publication Date: 2019-10-11
XI AN JIAOTONG UNIV
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Problems solved by technology

At present, there is no report on the stationary phase of membrane receptors with high expression of MrgprX2, a hypersensitive component.

Method used

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  • Recombinant cell with high expression of mrgprx2 and stationary phase of membrane receptor with high expression of mrgprx2 and its preparation method and application
  • Recombinant cell with high expression of mrgprx2 and stationary phase of membrane receptor with high expression of mrgprx2 and its preparation method and application
  • Recombinant cell with high expression of mrgprx2 and stationary phase of membrane receptor with high expression of mrgprx2 and its preparation method and application

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Embodiment Construction

[0031] The present invention will be further described in detail below in conjunction with embodiments and drawings.

[0032] The present invention constructs the eukaryotic expression vector LV5-MrgprX2 lentivirus of the full length of MrgprX2 gene and transfects HEK293 cells to obtain stable MrgprX2 high-expressing recombinant cells, which are recorded as HEK293-MrgprX2 cells, and HEK293- Based on MrgprX2 cells, a stationary phase with high expression of MrgprX2 membrane receptor was prepared. That is, the cell membrane of the recombinant cells with high expression of MrgprX2 is extracted and separated by a differential centrifugation method, and the separated cell membrane is physically adsorbed with activated silica gel to form a fixed phase of MrgprX2 high expression membrane receptor. Wet-pack the MrgprX2 high-expressing membrane receptor stationary phase with a column packing machine to obtain a cell membrane chromatography column for identifying allergic components acting...

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Abstract

The invention discloses an MrgprX2 high-expression reconstituted cell and MrgprX2 high-expression membrane receptor stationary phase and a preparation method and application thereof. The reconstituted cell uses HEK293 cells as host cells for transfecting expression carriers containing MrgprX2 full-length genes; the MrgPrX2 receptor macromolecules can be stably expressed on the cell membrane; in addition, the complete receptor biological activity is realized. The MrgprX2 high-expression membrane receptor stationary phase is manufactured by adsorbing cytomembranes of MrgprX2 high-expression reconstituted cells by activated silica gel; the application to the specific screening and identification of MrgprX2 triggered anaphylactoid ingredients can be realized. Through practical application, the condition that the sinomenine hydrochloride can be specifically combined with MrgprX2 is discovered. Meanwhile, the cell level allergic experiments prove that the sinomenine hydrochloride can stimulate mastocytes to release histamine and beta-hexosaminidase; the result proves that the MrgprX2 high-expression membrane receptor stationary phase can really be applied to the specific identification of the anaphylactoid ingredients.

Description

Technical field [0001] The invention belongs to the field of biotechnology, and relates to a MrgprX2 highly expressed recombinant cell and a MrgprX2 highly expressed membrane receptor stationary phase based on the recombinant cell to specifically recognize allergy-like components, and a construction method and application thereof. Background technique [0002] With the increasing clinical application of traditional Chinese medicine, safety "incidents" of traditional Chinese medicine occur from time to time, especially the clinical safety of traditional Chinese medicine injections. Allergy-like reactions are also called pseudoallergic reactions, which are exogenous stimuli that directly stimulate mast cells, causing them to release histamine-like active transmitters. Allergic reactions are generally not mediated by IgE and can be produced when the patient is first treated with medicine. Studies have shown that the G protein activated by allergic stimuli is MRGPRS (MAS-Related G Pr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/10C12N15/867C07K17/14G01N33/68
Inventor 贺浪冲张涛韩省力马维娜曹娇贺怀贞刘瑞车德路吕艳妮
Owner XI AN JIAOTONG UNIV
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