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Polypeptide and bacterin for preventing and curing infection of polytype human being papilomavirus

A use and antibody technology, applied in the field of immunology, can solve the problems that HPV has no preventive or therapeutic effect, the vaccine cannot target multiple types of HPV at the same time, and there are few cross-reactions between types

Active Publication Date: 2010-04-28
北京祥益华晟医疗器械有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But the disadvantage is that HPV16 L1 VLP is used as vaccine, the production cost is high, and it has no preventive or therapeutic effect on other types of HPV
However, its results have not been confirmed by other researchers' experiments and subsequent related reports, and its reaction is limited to two low-risk HPV types, and it cannot react with high-risk HPV types [15]
[0006] The second category is a vaccine composed of L1 and L2. Although its preventive effect is better than that of VLP composed of L1 alone, it has obvious disadvantages in terms of preparation and cost.
[0012] The biggest shortcoming of the existing HPV vaccine research results is that the designed vaccine cannot target multiple types of HPV at the same time, especially cannot induce immune responses against high-risk and low-risk HPV at the same time, or cannot target multiple types of HPV at the same time. Types of HPV Infection Produce Prophylaxis or Treatment
So far, due to the strong HPV type specificity and less cross-reaction between types, there is no HPV vaccine against multiple types used in clinical practice.

Method used

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  • Polypeptide and bacterin for preventing and curing infection of polytype human being papilomavirus
  • Polypeptide and bacterin for preventing and curing infection of polytype human being papilomavirus
  • Polypeptide and bacterin for preventing and curing infection of polytype human being papilomavirus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Preparation of specific peptide antiserum

[0065] (a) Synthesis of HPV L1 short peptide

[0066] In this example, based on the protein sequence of the HPV16L1448-477aa segment, in order to target multiple types of HPV, a G→A mutation was introduced at position 465.

[0067] The designed HPV L1 short peptide is 30 amino acids in length, and its sequence is as follows:

[0068] N-terminal-EVNLKEKFSADLDQFPLARKFLLQAGLKAK-C-terminal (SEQ ID NO: 1).

[0069] Beijing Saibaisheng Gene Technology Co., Ltd. was entrusted to synthesize the HPV L1 short peptide in the Genemed Synthesis Inc. company of the United States by conventional methods.

[0070] (b) Preparation of antiserum

[0071] Dissolve the short peptide synthesized above in PBS, add Furend.s complete adjuvant 1:1, emulsify and immunize Japanese long-eared white rabbits and Balb / c mice with doses of 25ug and 10ug, respectively, three times in total. On the 14th day of the last immunization, the animals were sacrifi...

Embodiment 2

[0073] ELISA Detection of Immune Specificity of Short Peptide Immune Antiserum

[0074] HPV16 virus particles, recombinant HPV16L1VLP, HPV6b L1VLP, HPV16 virus particle mouse antiserum, recombinant HPV16L1VLP and HPV6b L1VLP rabbit and mouse antisera were obtained from the Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences; Disease Research Institute. HRP-labeled and fluorescent-labeled secondary antibodies were purchased from Beijing Zhongshan Biotechnology Co., Ltd. (Sigma products). Both Caski cells and Hela cells were purchased from the China Center for Type Culture Collection (CCTCC) of Wuhan University.

[0075] Coat HPV16 virus, recombinant HPV16L1VLP, HPV6b L1VLP, Caski cultured cells containing HPV16 virus, and Hela cultured cell lysates containing HPV18 or their culture fluids on plastic plates, and use rabbit or mouse anti-short peptide antiserum to it react.

[0076] The result is as figure 1 shown. The short peptide immune antiserum of m...

Embodiment 3

[0078] Western blot detection of immunospecificity of short peptide immune antiserum

[0079] Caski cells, Hela cells, and insect cells cultured for 72 hours after transfection with HPV6b and 16L1 recombinant baculovirus were lysed, and the lysate was subjected to SDS polyacrylamide gel electrophoresis. anti.

[0080] The result is as figure 2 shown. In the range of protein molecular weight about 56K, there is a positive reaction band.

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Abstract

This invention describes a polypeplide that can simultaneously induce immunity to a wide variety of types, especially to high-risk and low-risk types, and the vaccines containing this polypeplide. Thevaccines in this invention can prevent many types of HPV infection and / or treat a fraction of HPV infective diseases.

Description

technical field [0001] The invention belongs to the field of immunology, and in particular relates to a polypeptide and a vaccine for the prevention and treatment of polytype human papillomavirus (HPV) infection, and the polypeptide is derived from HPV coat protein 1. Background technique [0002] Human papillomavirus (HPV) is a kind of DNA virus with strict host range and tissue specificity, and more than 100 types have been found. Among them, HPV16, 18, 31, 33, 45, and 58 are a main factor causing digestive tract and reproductive tract cancer, especially cervical cancer; HPV6 and 11 are important pathogenic factors causing sexually transmitted diseases such as genital warts [1-5] . In order to effectively prevent the infection and spread of this virus, countries all over the world are actively developing HPV vaccines. [0003] HPV vaccines can be roughly divided into two categories: therapeutic vaccines and preventive vaccines. Since HPV cannot be cultured by tissue and ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/025C07K16/08A61K38/16A61K39/395A61P31/20
Inventor 肖长义万涛黄利鸣袁太宁
Owner 北京祥益华晟医疗器械有限责任公司