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Protective HPV (Human Papilloma Virus) immunoglobulin of yolk (IgY) and application thereof

A yolk antibody, HPV16L1 technology, applied to the yolk antibody of HPVL1 recombinant microcircular DNA, the application field of prevention/treatment of HPV infection, can solve the problem of no effective treatment method for HPV virus, etc., achieves low production cost, high antibody titer, Simple preparation process

Inactive Publication Date: 2018-08-14
江苏润洁生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no precedent for HPV L1 yolk antibody obtained by immunization with nucleic acid antigen to prevent cervical cancer
[0007] At present, there is no effective treatment for intracellular HPV in cervical mucosa

Method used

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  • Protective HPV (Human Papilloma Virus) immunoglobulin of yolk (IgY) and application thereof
  • Protective HPV (Human Papilloma Virus) immunoglobulin of yolk (IgY) and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Example 1 Preparation of yolk antibody

[0067] 1. Preparation of recombinant minicircle DNA

[0068] 1) Design and synthesize the HPV-L1 recombinant fragment with BglII and SalI restriction sites added at both ends to obtain the target gene;

[0069] 2) The above-mentioned target gene fragment and the ZY781 vector are simultaneously double-cut with BglII and SalI, and the target fragment is cloned into the multiple cloning site of the vector ZY781 according to the conventional method after the gel is recovered, and the recombinant plasmid is obtained, which is the parental plasmid (PP) ;

[0070] 3) Transform Escherichia coli ZYCY10P3S2T with the above-mentioned parental plasmid, culture it overnight, after extracting the plasmid, the plasmid is digested with enzyme electrophoresis and plasmid sequencing to verify that the correct fragment is inserted;

[0071] 4) Put the above-sequenced Escherichia coli into TB culture medium containing 50μg / ml kana, and cultivate overnight at...

Embodiment 2

[0084] Example 2 Gel for prevention / treatment of HPV infection and its effect evaluation

[0085] 1. Ingredients: HPV L1 chicken egg yolk antibody (containing nano liposomes), PHMB, lactose, glycerin, distilled water.

[0086] Use vaginal gel formulations to infuse patients with highly active anti-HPV antibodies. Nanoliposomes are used to achieve transmembrane transport of antibodies, so that the antibodies can directly neutralize the HPV virus in the cell, which solves the current clinical problem of HPV in cervical mucosa cells. There is no effective cure for the virus. At the same time, the yolk antibody can also neutralize the newly released virus in the infected area, making it lose the ability to transmit it through sexual contact.

[0087] The pH is 5.0, which is weakly acidic, and the vaginal environment is also acidic to avoid damage to the homeostasis of the vaginal environment.

[0088] In addition to vaginal gel, the chicken egg yolk antibody of the present invention can ...

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Abstract

The invention belongs to the field of biomedicines and particularly relates to an immunoglobulin of yolk (IgY) of HPV (Human Papilloma Virus) L1 recombinant minicircle DNA (DeoxyriboNucleic Acid) andapplication of the IgY in preventing / treating HPV infection, wherein the HPV L1 recombinant minicircle DNA contains an L1 gene fragment of HPV16 and a signal peptide sequence, and is used as a nucleicacid antigen for directly immunizing laying hens to obtain the IgY disclosed by the invention. The biological activity of the IgY obtained by immunizing the laying hens through DNA nucleic acid antigen intramuscular injection is far superior to that of IgY prepared by a general technique (subcutaneous injection of a protein antigen), the antibody titer is higher and the specificity is higher. TheIgY disclosed by the invention can be used for preparing a drug for preventing / treating the HPV infection; a high-activity anti-HPV antibody is inputted into HPV-positive patients by vaginal medication; by combining with a nano-liposome method, an antibody transmembranely enters the cell and directly neutralizes an HPV virus in cells. The IgY disclosed by the invention is of great significance for preventing cervical cancer.

Description

Technical field [0001] The invention belongs to the field of biomedicine, and specifically relates to a yolk antibody of HPV L1 recombinant minicircle DNA and its application in the prevention / treatment of HPV infection. Background technique [0002] Cervical cancer accounts for the second place among female gynecological malignancies, and has become an important disease threatening women’s health. Nearly 50,000 people die of cervical cancer in China each year. Research on the cause of cervical cancer has always been paid attention to by scholars at home and abroad. The key to cervical cancer is HPV infection. HPV belongs to the papillomavirus genus of the papillomavirus family and is a spherical, double-stranded circular DNA virus. HPV mainly infects humans through direct or indirect contact with contaminated items or through sexual transmission. HPV infection is host-specific and tissue-specific, and it can only infect human skin and mucous membrane damaged basal cells. [000...

Claims

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Application Information

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IPC IPC(8): C12N15/62C12N15/85C07K16/08C07K16/02A61K39/42A61K9/127A61K31/785A61P31/02A61P31/20A61P35/00
CPCA61K9/0034A61K9/127A61K31/785A61K39/42A61K2039/505A61P31/02A61P31/20A61P35/00C07K14/005C07K16/02C07K16/084C07K2317/11C07K2319/02C07K2319/21C12N15/85C12N2710/20022C12N2800/106A61K2300/00
Inventor 庞以芹谢亦武吴必杰魏勇
Owner 江苏润洁生物科技有限公司
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