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Mi-2 recombinant antigen as well as preparation method and application thereof

A recombinant antigen and mi-2 technology, applied in biochemical equipment and methods, recombinant DNA technology, chemical instruments and methods, etc., can solve the problems of inability to meet the performance of diagnostic reagents, low immunogenicity, etc., and improve the space position The effect of overcoming the problem of resistance, overcoming low immunogenicity, and overcoming the difficulty of extraction

Active Publication Date: 2020-10-30
ZHUHAI LIVZON DIAGNOSTICS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The Mi-2 antigen protein is composed of 1,912 amino acids, and the relative molecular weight of the protein is relatively large. Although the full-length Mi-2 antigen protein recombinantly expressed by insect cells is easy to obtain, its immunogenicity is low, which cannot meet the performance of diagnostic reagents.

Method used

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  • Mi-2 recombinant antigen as well as preparation method and application thereof
  • Mi-2 recombinant antigen as well as preparation method and application thereof
  • Mi-2 recombinant antigen as well as preparation method and application thereof

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Embodiment 1

[0056] Through the bioinformatics analysis of the antigenic epitope of Mi-2 antigen protein (UniProtKB-Q14839(CHD4_HUMAN), 1912aa), Mi-2 was divided into 8 segments and expressed in Escherichia coli respectively. The corresponding amino acid sequences are SEQ ID NO.1 (amino 1-239), SEQ ID NO.2 (amino 240-448), SEQ ID NO.3 (amino 449-677), SEQ ID NO.4 (amino 678-948) , SEQ ID NO.5 (amino 949-1209), SEQ ID NO.6 (amino 1210-1498), SEQ ID NO.7 (amino 1499-1696) and SEQ ID NO.8 (amino 1697-1912).

[0057] Plasmid construction and protein expression

[0058] [1] The primers corresponding to the 8 fragments are as follows:

[0059] The primers of the fragment of SEQ ID NO.1 are:

[0060] F1: 5'-atggcgtcgggcctgggc-3' (SEQ ID NO.11);

[0061] R1: 5'-gctctccaccacagctac-3' (SEQ ID NO.12);

[0062] The primers of the fragment of SEQ ID NO.2 are:

[0063] F2: 5'-atggtgacagccactgaggttg-3' (SEQ ID NO.13);

[0064] R2: 5'-atggtggtcatcctcctcttcg-3' (SEQ ID NO.14);

[0065] The primers o...

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Abstract

The invention relates to the technical field of biology, and particularly provides a Mi-2 recombinant antigen as well as a preparation method and application thereof. The Mi-2 recombinant antigen comprises at least one sequences representing by SEQ ID NO.1, SEQ ID NO.5 or SEQ ID NO.7, and does not contain sequences representing by SEQ ID NO.2 and / or SEQ ID NO.6. The Mi-2 recombinant antigen improves the sensitivity and specificity of Mi-2 antibody detection, is used for Mi-2 antibody detection or polymyositis diagnosis, can greatly improve the accuracy, and provides a new technical support forproduction and application.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a Mi-2 recombinant antigen and its preparation method and application. Background technique [0002] Polymyositis (PM) is a common autoimmune disease, and Mi-2 antibody is a kind of myositis-specific antibody, which is of great significance in the diagnosis of myositis. The target antigen of the anti-Mi-2 antibody is the Mi-2 antigen protein with a relative molecular mass of 218kD located in the nucleoplasm. [0003] The common way to obtain natural Mi-2 antigen is to extract it from rabbit thymus, which is difficult to extract and has a low yield. The Mi-2 antigen protein is composed of 1,912 amino acids, and the relative molecular weight of the protein is relatively large. Although the full-length Mi-2 antigen protein recombinantly expressed in insect cells is easy to obtain, its immunogenicity is low and cannot meet the performance of diagnostic reagents. . [0004] In view of ...

Claims

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Application Information

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IPC IPC(8): C12N9/00C12N15/52C07K19/00C12N15/70G01N33/68G01N33/573C12R1/19
CPCC12N9/00C12N15/70G01N33/6854G01N33/6893G01N33/573C07K2319/00G01N2800/10
Inventor 胡志强况承钰曾敏霞张晨光
Owner ZHUHAI LIVZON DIAGNOSTICS
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