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Preparation method of anti-HPV infection antibody

An antibody and antigen technology, applied in chemical instruments and methods, antiviral immunoglobulin, egg-derived immunoglobulin, etc., can solve the problems of heterologous microbial contamination, genetic instability of antigen-expressing strains, and low yield of antibody preparation. , to prevent infection

Inactive Publication Date: 2016-08-17
LANJIATANG BIOLOGICAL MEDICINE FUJIAN CO LTD
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  • Abstract
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Problems solved by technology

[0004] According to the current technological development, there are some technical bottlenecks in the preparation of active and passive immunization antigens, such as: genetic instability of antigen expression strains, contamination of heterologous microorganisms during fermentation, instability of purification yield, low yield of antibody preparation, etc.

Method used

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  • Preparation method of anti-HPV infection antibody
  • Preparation method of anti-HPV infection antibody
  • Preparation method of anti-HPV infection antibody

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preparation example Construction

[0021] The technical solution adopted in the present invention is: providing a method for preparing an antibody against HPV infection, comprising the following steps: passing human heparan sulfate proteoglycan, furin-furin and capsid protein of pathogenic subtype of HPV through Prokaryotic expression, yeast or CHO expression preparation, or through the preparation of virus-like particles, the above-mentioned key proteins of virus infection, including virus receptor, auxiliary protease and virus capsid protein, are used as antigens to immunize animals to obtain antibodies against HPV infection.

[0022] Preferably, in the above method for preparing an antibody against HPV infection, the animal is a laying hen.

[0023] Preferably, in the preparation method of the above-mentioned anti-HPV infection antibody, the heparan sulfate-like proteoglycan is prepared through prokaryotic expression and yeast / CHO expression, and a relatively pure target protein is obtained after separation a...

Embodiment 1

[0027] The chicken egg yolk immunoglobulin (antibody) preparation of embodiment one anti-HPV-16 infection

[0028] 1 Gene cloning and protein separation and purification

[0029] 1.1 Query the gene sequence of related protein molecules through NCBI / Genebank, including: HPV (low-risk type, high-risk type) L1, L2 full sequence, heparan sulfate proteoglycan and furin-furin, and obtain the complete gene through gene synthesis sequence.

[0030] Wherein: HPV type-16 L1 gene sequence: (SEQ ID NO: 1).

[0031] HPV type-16 L2 gene sequence: (SEQ ID NO: 2).

[0032]1.2 After the L1 and L2 of the above-mentioned heparan sulfate proteoglycans, furin- and HPV-16 subtypes are synthesized by genes, they are cloned into prokaryotic expression vectors or eukaryotic expression vectors (or have SV40 promoter and Eukaryotic enhancers, and appropriate reporter genes such as HRP, fluorescent proteins, etc.). The genetic engineering steps include: designing PCR primers according to the gene seq...

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Abstract

The invention relates to the technical field of biopharmaceutics, in particular to a preparation method of an anti-HPV infection antibody, comprising the following steps: transforming capsid protein of subtype of HPV pathogenicity into 293TT cells, packaging pseudovirus, preparing virus-like particles and using the virus-like particles as part of antigen to immunize animals to acquire an anti-HPV infection antibody. The antibody obtained by the preparation method of the anti-HPV infection antibody is a polyclonal specific antibody of certain purity and concentration obtained by immunizing laying hens (or other bird species) with key proteins of invasion in an HPV infection course, namely HPV L-1 / L2(17-36 short peptides), furin and heparan sulfate proteoglycan and adding proper adjuvant through an antibody engineering technique, and the antibody can quickly neutralize HPV and block the HPV invasion passage to play the effect of preventing high risk type HPV infection.

Description

technical field [0001] The invention relates to the technical field of biopharmaceuticals, in particular to a method for preparing an antibody against HPV infection. Background technique [0002] Human papillomavirus (HPV) mainly causes verrucous lesions on the skin and mucous membranes. According to its relationship with tumorigenesis, HPV can be divided into high-risk types and low-risk types, among which high-risk HPV infection has been proved to be the main cause of genital cancer including female cervical cancer; low-risk type mainly causes genital warts. Cervical cancer is known as the "beauty killer". Among women aged 25-45, the mortality rate of cervical cancer ranks second. According to the statistics of the World Health Organization, there are 529,000 new cases of cervical cancer every year in the world, and about 250,000 people die from cervical cancer, of which more than 90% are from developing countries. [0003] With the in-depth research on molecular biology...

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Application Information

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IPC IPC(8): C07K16/08C07K16/02
CPCC07K16/082C07K16/02
Inventor 胡博志邹潮兰增金
Owner LANJIATANG BIOLOGICAL MEDICINE FUJIAN CO LTD
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