Application of nachr-α1-ecd protein in medical detection
A 1-ECD, protein technology, applied in the field of biochemistry, can solve the problems of difficult separation, low yield, complicated operation, etc.
Active Publication Date: 2020-10-23
SHANGHAI CHANGHAI HOSPITAL
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Problems solved by technology
[0003] However, there is currently no human AChR-Ab clinical detection kit for sale on the market, and the old method continues to use animal or human muscle extraction, which is complex and costly, and cannot meet the needs of a large number of patients in reality.
However, directly according to the amino acid sequence of human nAChR for gene expression, although the target protein can be obtained, it is often difficult to separate or the purity of the separation is not high and the yield is low due to the nature of the target protein.
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[0038] In order to make the technical means, creative features, goals and effects of the present invention easy to understand, the following examples are combined with the accompanying drawings to describe the nAChR-α1-ECD protein of the present invention and its preparation method and the application of the protein in medical testing. elaborate.
[0039] The reagents presented in the examples were purchased commercially unless the sources were indicated otherwise.
[0040] IPTG, isopropyl-β-D-thiogalactopyranoside.
[0041] LB medium, Luria-Bertani medium.
[0042] mM means mmol / L.
[0043] Tris is tris(hydroxymethyl)aminomethane buffer solution.
[0044] Gua-HCl is guanidine hydrochloride.
[0045] M is mol / L.
[0046] EDTA stands for ethylenediaminetetraacetic acid.
[0047] DTT is dithiothreitol.
[0048] WB is the abbreviation of Western blot.
[0049] Tween, Tween, polysorbate, the following values indicate that the types of fatty acids combined with polymerize...
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Abstract
The invention provides an application of nAChR-alpha 1-ECD protein in medical detection. The application is characterized in that the amino acid sequence of the protein is SEQ ID 1, and the protein has the molecular weight of 28.7 kDa and the isoelectric point of 6.0. Besides, the invention further provides a specific method for detecting whether nAChR-Ab exists in patient serum or not or furtherdetecting the concentration of the nAChR-Ab by use of the nAChR-alpha 1-ECD protein with an ELISA indirect method and a WB (western blotting) method. As Escherichia coli expressed nAChR-alpha 1-ECD protein is used, the components are clear, the purity is high, and the detection result is reliable and good in repeatability. The detection cost is low due to the fact that the protein is artificiallyprepared.
Description
technical field [0001] The invention relates to the application of nAChR-α1-ECD protein in medical detection and belongs to the field of biochemistry. Background technique [0002] The Department of Neurology of Shanghai Changhai Hospital began to carry out clinical trial research on myasthenia gravis (MG) in 1978. It used electric rays and human muscle to extract nAChR antigen and detected myasthenia gravis nAChR antibody (nAChR-Ab). This method had to be discontinued due to the increasingly limited materials. [0003] However, there is currently no human AChR-Ab clinical detection kit for sale on the market, and the old method continues to use animal or human muscle extraction. The operation is complicated and costly, and it cannot meet the needs of a large number of patients in reality. However, gene expression after artificial synthesis directly based on the amino acid sequence of human nAChR, although the target protein can be obtained, is often difficult to separate o...
Claims
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IPC IPC(8): G01N33/68
CPCG01N33/6887G01N33/6893G01N2333/4712G01N2800/28
Inventor 陈园园涂来慧毕晓莹刘海平张秀天刘善荣
Owner SHANGHAI CHANGHAI HOSPITAL