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Linear epitope minimum motif peptide of human papilloma virus type 58 L1 protein and application thereof

A technology of L1 protein and epitope peptide, which can be applied in the directions of viral antigen components, antiviral agents, and medical preparations containing active ingredients, etc., which can solve the problem of inability to identify linear antigenic epitopes.

Inactive Publication Date: 2012-09-26
广东南湾医药技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the limitations of the previous epitope identification methodology, it was impossible to identify all linear epitopes on a target protein, especially the minimal motif (which can improve the accuracy of this detection, and avoid the formation of normal human serum. Thousands of unknown antigens and antibodies have cross-reactions), so far there is no specific and sensitive serological detection method for HPV infection, although in the early days, many serums from HPV-infected patients were used to find specific markers for serological detection of HPV infection (Dillner J, et al. al. Int J Cancer, 45: 529-535, 1990; Dillner J, et al. Int J Cancer, 46: 703-711, 1990; Bleul C, et al. J Clin Microbiol, 29: 1579-1588, 1991; Jenison S, et al. J Virol, 65: 1208-1218, 1991)

Method used

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  • Linear epitope minimum motif peptide of human papilloma virus type 58 L1 protein and application thereof
  • Linear epitope minimum motif peptide of human papilloma virus type 58 L1 protein and application thereof
  • Linear epitope minimum motif peptide of human papilloma virus type 58 L1 protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Linear BCE scanning mapping of human papillomavirus type 58 (HPV58) L1 protein

[0034] Materials and methods:

[0035] 1. HPV58 gene clone (pLink322 / HPV58 plasmid) is commercially available. The pRSET C plasmid was purchased from Invitrogen, USA. The heat-inducible expression plasmid pXXGST-1 was constructed by the inventor of this patent (patent application number: 200710173305.2). Escherichia coli BL21 (DE3) strain was commercially available.

[0036] 2. Restriction enzymes EcoR I, BamH I, Sal I, Taq enzyme and T4 DNA ligase were purchased from Japan TaKaRa Biotechnology Company, pre-stained protein molecular weight standards and horseradish peroxidase-labeled goat anti-rabbit secondary antibody (IgG / HRP) Shanghai Sangon Biotechnology Service Company. ECL chemiluminescent color development kit was purchased from GE Healthcare UK. A 0.2 μm nitrocellulose membrane was purchased from Whatman Gmbh Hahnestr. 6 × His monoclonal antibody was purchased from...

Embodiment 2

[0048] Embodiment 2: HPV58 viral L1C protein P27 epitope peptide minimum motif identification

[0049] Materials and methods:

[0050] See the corresponding part of Example 1.

[0051] The specific steps for identifying the minimal motif of the P27 epitope peptide are as follows:

[0052] 1. According to the results of scanning and positioning the epitope of the L1C protein antigen in Example 1, the minimal motif identification of the P27 epitope peptide was carried out as an example. Design a series of 8 peptides (P139-P149) that span the 18 peptide sequences of the P27 epitope peptide and overlap each other by 7 amino acid residues (P139-P149) to encode the positive and negative strands of DNA (5'-end plus 5'-gatcc, 3'-end plus taag-3'; minus-strand 5'-end plus 5'-tcgactta, 3'-end plus g-3'), send out DNA synthesis.

[0053] The 2-4 operation steps are the same as the 2-4 steps in the embodiment 1.

[0054] 5. The blotted membrane was repeatedly washed four times with P...

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Abstract

The invention belongs to the technical field of biomedicine and bioinstrumentation, in particular to an epitope (Bcellepitope) minimum motif peptide of human papilloma virus (HPV) type 58 L1 structural protein and application thereof. The invention provides 16 amino acid sequences containing minimum motif peptide (including 8 peptide sequence capable of cutting off expression of carrier protein such as GST188), and the amino acid sequences can be taken as antigen for independently or combinedly specific detection of blood serum of HPV type 58 virus infection patient and can be used for developing preventive HPV type 58 multi-epitope peptide vaccine capable of simultaneously inducing humoral immunity. The amino acid sequences of HPV type 58 L1 epitope minimum motif peptide and short peptide are shown as SEQIDNo.1-SEQIDNo.32.

Description

Technical field [0001] The present invention belongs to the field of biomedicine and biological testing technology, and it involves all linear B cells (B CELL EPITOPE, BCE, or Bcetop, BCE, orIt is called the application of antigen tables or decision clusters) and its minimum base -order peptide, as well as the application of these surface -based basis peptides. Background technique [0002] Chemical synthetic peptide vaccines have been expected to be the third milestone after the history of human application vaccine development in the history of human application vaccine and virus sub -unit vaccine.However, the development of preventive synthetic peptide vaccines composed of one or two tablets lacks full and effective protection. Since the 1970s, there have been no clinical application of the approved person.Therefore, in order to achieve the effectiveness of protection of human clinical application, the synthetic peptide vaccine should be used to combine as many linear antigen t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K5/113C07K5/117C07K5/11C07K5/083C07K5/103C07K7/06C07K2/00A61K39/295A61P31/20A61P35/00G01N33/569A61K39/12
Inventor 徐万祥顾少华何亚萍季朝能唐海平王健孙志达谢毅
Owner 广东南湾医药技术有限公司
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