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HPV egg yolk antibody and application thereof in preparing drug for treating HPV infection

A technology of egg yolk antibody and medicine, which is applied in the application field of HPV egg yolk antibody and medicine, can solve the problems of no obvious therapeutic effect, and achieve the effect of low production cost, high antibody titer and strong specificity

Active Publication Date: 2019-04-23
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 E6 / E7 yolk antibodies obtained by immunization with nucleic acid antigens for the treatment of HPV infection.
[0007] The current preventive vaccine Gardasil / Cervarix can effectively prevent HPV infection, but it has no obvious therapeutic effect on patients with existing HPV infection or precancerous lesions, so it is necessary to develop therapeutic drugs against HPV infection

Method used

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  • HPV egg yolk antibody and application thereof in preparing drug for treating HPV infection
  • HPV egg yolk antibody and application thereof in preparing drug for treating HPV infection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] The preparation of embodiment 1 yolk antibody (IgY)

[0070] 1. Preparation of recombinant microcircle DNA

[0071] 1) designing and synthesizing the HPV E6 / E7 recombinant fragment shown in SEQ ID NO: 8 with BglII and SalI restriction sites added at both ends to obtain the target gene;

[0072] 2) Digest the above target gene fragment and the ZY781 vector with BglII and SalI at the same time, and clone the target fragment into the multi-cloning site of the vector ZY781 according to the conventional method after gel recovery to obtain a recombinant plasmid, which is the parental plasmid (PP) ;

[0073] 3) Transform Escherichia coli ZYCY10P3S2T with the above-mentioned parental plasmid, culture overnight, extract the plasmid, and then verify the insertion of the correct fragment by electrophoresis and sequencing of the plasmid;

[0074] 4) Inoculate the Escherichia coli with correct sequencing above into TB medium containing 50 μg / ml kana, culture overnight at 37°C and ...

Embodiment 2

[0083] Embodiment 2 treats the gel 1 that HPV infects

[0084] 1. Preparation of Liposome Microcapsules

[0085] ① Dry the specific IgY obtained in Example 1 with a lyophilizer to make a dry powder. Add it into an ultrafine pulverizer for grinding and grinding, and process it into ultrafine particles with a size between 1-100nm and a particle size of more than 15,000 meshes to obtain nano-sized antibodies.

[0086] ② Dissolve lecithin and cholesterol in ether, then add nano-specific IgY to 4mmol / L phosphate buffered saline (PBS) to make IgY solution, sonicate for 2min (each treatment 0.5min, intermittent 0.5min), immediately Evaporate under reduced pressure in a water bath until it becomes a gel, vortex to invert the phase of the gel, continue to evaporate to remove the ether, and then ultracentrifuge (35000r / min, 30min) to separate and remove the unencapsulated IgY, and wash the precipitate with water for two Once, centrifuged to obtain a precipitate, which was diluted with...

Embodiment 3

[0091] Embodiment 3 treats the gel 2 that HPV infects

[0092] 1. Preparation of Liposome Microcapsules

[0093] Same as Example 2.

[0094] 2. Gel Preparation

[0095] In parts by weight, 15 parts of liposome microcapsules, 40 parts of glycerin, 22 parts of water, 2.5 parts of lactose, 3 parts of sucrose, and 0.2 parts of sodium citrate were mixed and adjusted to pH 5.0.

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Abstract

The invention belongs to the field of biomedicine, and particularly relates to an egg yolk antibody of HPV E6 / E7 recombined minicircle DNA and application of the antibody in preparing a drug for treating HPV infection. The egg yolk antibody is obtained by utilizing HPV E6 / E7 recombined minicircle DNA as a nuclear acid antigen for direct immunization of a laying hen. The egg yolk antibody can be used for preparing the drug for treating HPV infection. The egg yolk antibody is prepared into freeze-dried powder and subjected to superfine grinding, after lipidosome encapsulating, a nano-liposome encapsulating the egg yolk antibody is prepared, the antibody enters cells in a transmembrane mode, the high-activity anti-HPV antibody is input to a patient with positive HPV, and E6 / E7 protein in an HPV virus inside the cells is directly neutralized. The antibody is combined with antibacterial active ingredients (antibacterial compounds or gynecologic inflammation pathogen specific antibodies) andacceptable pharmaceutical carriers to be prepared into vagina spraying agents, vagina washing liquor, vagina gels, vagina soft and hard capsules, vagina suppositories, vagina membranes, vagina tablets, effervescent tablets, ointment, creams and the like, and can also be prepared into lozenges, chewable tablets, chewing gums and the like for both male and female.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to an HPV egg yolk antibody and its application in preparing a medicine for treating HPV infection. Background technique [0002] Cervical cancer accounts for the second place among female gynecological malignancies, and has now become an important disease that threatens women's health. In China, nearly 50,000 people die from cervical cancer every year. The etiology of cervical cancer has always been valued by scholars at home and abroad. The key to cervical cancer is HPV infection. HPV is a spherical, double-stranded circular DNA virus belonging to the genus Papillomavirus of the family Papillomaviridae. HPV infects humans mainly through direct or indirect contact with contaminated objects or through sexual transmission. HPV infection has host specificity and tissue specificity, and can only infect damaged basal cells of human skin and mucous membranes. [0003] The damage...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/62C12N15/85C07K16/08C07K16/02A61K39/42A61K9/127A61K31/785A61P31/02A61P31/20A61P35/00
CPCA61K9/0034A61K9/127A61K31/785A61K2039/505A61P31/02A61P31/20A61P35/00C07K14/005C07K16/02C07K16/084C07K2317/11C07K2319/02C07K2319/21C12N15/85C12N2710/20022C12N2800/106
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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