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Linear epitope minimum motif peptide of human papilloma virus type 58 E7 protein

An antigen peptide, the smallest technology, applied in the direction of virus antigen components, antiviral agents, peptides, etc., can solve the problems that have not been developed

Inactive Publication Date: 2013-06-19
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, previous studies on E6 and E7 epitopes were mainly focused on HPV16 and HPV18 (Dillner J, et al. Int J Cancer, 45: 529-535, 1990; Bleul C, et al. J Clin Microbiol, 29: 1579- 1588, 1991; Gao L, et al. J Gen Virol, 75: 157-164, 1994), while the identification of the linear epitope of HPV58 E7 protein has not been carried out

Method used

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  • Linear epitope minimum motif peptide of human papilloma virus type 58 E7 protein
  • Linear epitope minimum motif peptide of human papilloma virus type 58 E7 protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Example 1: Linear BCE scanning mapping of human papillomavirus type 58 (HPV58) E7 protein

[0026] Materials and methods:

[0027] 1. HPV58 gene clone (pLink322 / HPV58 plasmid) was purchased from Dr. Matsukura T, Japan. 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 preserved by the State Key Laboratory of Genetic Engineering, Fudan University.

[0028] 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 fro...

Embodiment 2

[0040]Example 2: Identification of minimal motifs of human papillomavirus type 58 (HPV58) E7 protein P15 epitope peptide

[0041] Materials and methods:

[0042] See the corresponding part of Example 1.

[0043] The specific steps for identifying the minimal motif of the P15 epitope peptide are as follows:

[0044] 1. According to the results of E7 protein antigen epitope scanning and positioning in Example 1, the minimum motif identification was carried out by taking the E7-8 epitope peptide P15 as an example. Design a series of 8 peptides that span the P15 epitope peptide with 14 peptide sequences overlapping each other by 7 amino acid residues to encode positive and negative strands of DNA (5'-end plus 5'-gatcc, 3'-end plus taag-3'; Add 5'-tcgactta to the 5'-end of the negative strand, add g-3' to the 3'-end), and send out DNA synthesis. After identifying the only imprinted positive P66 peptide in the P60-P66 series of 8 peptides, design and synthesize short peptides (3-...

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Abstract

The invention belongs to the technical field of bio-medicine and biological detection, in particular to a bcellepitope minimum motif peptide of human papilloma virus (HPV) type 58 oncogenic E7 protein and a use thereof. The invention provides eight epitope peptides containing minimum motif (including the 8-peptide sequence which can be in protein expression by truncated GST188 and other carriers), which can be used as antigens alone or be combined for specific detection of serum of a patient infected with HPV type 58 virus, and a specific minimum motif peptide amino acid sequence of a therapeutic HPV 58 multi-epitope peptide vaccine, which is used for developing and simultaneously inducing humoral immunity. The amino acid sequences of the HPV type 58 E7 epitope minimum motif peptide and a short peptide are respectively shown in SEQ ID No.1-SEQ ID No.16.

Description

technical field [0001] The invention belongs to the technical field of biomedicine and biological detection, and specifically relates to all linear B cell epitopes (B cell epitope, BCE, or called antigenic determinants) and their minimal motif peptides, and the application of these epitope motif peptides. Background technique [0002] Human papillomavirus (HPV) is a general term for a group of viruses that can infect epithelial tissues such as the female reproductive tract. It is known that more than 100 HPV viruses encode 6 early proteins of E1, E2, E4-E7 and 2 late proteins of L1 and L2 (Stanley MA. Human papillomavirus vaccines. Rev Med Virol, 16: 139-149, 2006) The E7 protein encoded by high-risk HPV (such as HPV16 and HPV18, etc.), which can lead to cervical cancer in women, has been proved to be highly carcinogenic. Therefore, the E7 protein of high-risk HPV is one of the important targets for developing immunotherapy for cervical cancer (Preville X, et al. Cancer Re...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K5/113C07K7/06C07K2/00A61K39/295A61P31/20A61P35/00G01N33/569A61K39/12
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