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Recombinant rubella virus E1 protein and uses thereof

A rubella virus and protein technology, applied in the field of genetic engineering, can solve problems such as the difficulty in the separation and purification of antigenic component E1, the lack of research and development of commercial diagnostic kits, and the difficulty in the separation of antigenic components, so as to meet the needs of clinical diagnosis and sensitivity High and specific effect

Inactive Publication Date: 2011-05-11
吴丽霞 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the separation of the E1 / E2 / C antigenic components of rubella virus is relatively difficult, especially the separation and purification of the main antigenic component E1, which is basically not used in the research and development of commercial diagnostic kits

Method used

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  • Recombinant rubella virus E1 protein and uses thereof
  • Recombinant rubella virus E1 protein and uses thereof
  • Recombinant rubella virus E1 protein and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Preparation of recombinant rubella virus E1 protein

[0034] The amino acid sequence of the recombinant rubella virus E1 protein is designed according to the amino acid sequence of RV envelope glycoprotein E1 (glycoprotein E1), and the sequence is 203-295 amino acids of the amino acid sequence of RV envelope glycoprotein E1 (glycoprotein E1).

[0035] 1.1 Synthesis of recombinant DNA of rubella virus E1 protein

[0036] The amino acid sequence of RV envelope glycoprotein E1 (glycoprotein E1) is analyzed by computer, combined with the confirmed antigenic epitope, and a part of the sequence with strong antigenicity is selected.

[0037] RV包膜糖蛋白E1(glycoprotein E1)氨基酸全序列如下:EEAFTYLCTAPGCATQAPVPVRLAGVRFESKIVDGGCFAPWDLEATGACICEIPTDVSCEGLGAWVPAAPCARIWNGTQRACTFWAVNAYSSGGYAQLASYFNPGGSYYKQYHPTACEVEPAFGHSDAACWGFPTDTVMSVFALASYVQHPHKTVRVKFHTETRTVWQLSVAGVSCNVTTEHPFCNTPHGQLEVQVPPDPGDLVEYIMNYTG NQQSRWGLGSPNCHGPDWASPVCQRHS PDCSR LVGATPE RPRLRLVD ADDPL LR TAPGPGE VWVTP...

Embodiment 2

[0078] Example 2 Preparation and Performance Test of the Kit for Detecting Rubella Virus IgM Antibody Infection

[0079] 2.1 Preparation of a kit for detecting rubella virus IgM antibody infection

[0080] The recombinant rubella virus E1 protein prepared in Example 1 was used as the substrate for the preparation of the enzyme-labeled antigen of the kit, and the RVIgM antibody was detected by ELISA.

[0081] The kit was developed and used as follows:

[0082] 1) The principle of the kit: this strain uses anti-human IgM (μ chain) coated microwell plate, horseradish peroxidase (HRP) labeled recombinant rubella virus E1 as tracer, TMB chromogenic system, application capture The principle of the method is to detect the anti-rubella virus IgM antibody in human serum or plasma.

[0083] 2) The main components of the kit:

[0084] a) Pre-coated plates:

[0085] b) Coating solution: 0.05mol / l, carbonate buffer system with pH9.6

[0086] c) Blocking solution: composed of 0.01mol / L...

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Abstract

The invention relates to a recombinant rubella virus E1 protein and an application thereof. The adopted technical proposal is as follows: the amino acid sequence of the recombinant rubella virus E1 is shown in SEQ ID NO: 2, a reagent kit which is prepared by the recombinant rubella virus E1 protein and used for detecting the rubella virus IgM antibody infection is adopted, compared with the similar reagent kits on the market, the reagent kit has the advantages of strong specificity, high sensitivity, and the like, thereby well meeting the needs of clinical diagnosis of rubella virus infection. The invention further provides the application of the recombinant rubella virus E1 protein in the preparation of monoclonal antibodies, polyclonal antibodies and protein chips.

Description

Technical field: [0001] The invention relates to the field of genetic engineering, in particular to a recombinant rubella virus E1 protein and its application. Background technique: [0002] Rubella virus (rubella virus, RV) RV is the only member of the Rubellavirus genus in the Togaviridae family, and humans are its only natural host. Rubella itself is not serious, and acquired infection mainly manifests as acute fever, submandibular lymph node enlargement and other symptoms. The greatest threat to RV is its teratogenicity. Pregnant women infected with RV can cause congenital damage to the fetus, especially if infected with RV in the first 3 months of pregnancy, it can infect the fetus through the placenta, resulting in miscarriage, stillbirth, or a series of organ malformations in the newborn, that is, congenital rubella syndrome. Establishing a rapid and accurate detection method for RV infection is the key to preventing RV infection. [0003] At present, the diagnosti...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/40C07K14/19C12N15/63C12N1/21G01N33/68C07K16/10C12R1/19
Inventor 吴丽霞赵玉红于庭包洪王贵全李凤娣姜禾董亚俊徐田雪
Owner 吴丽霞
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