110 results about "Human papillomavirus types" patented technology

The invention provides an sgRNA combined with immunogene to inhibit high risk HPV expression, a gene knockout vector thereof, and an application of the sgRNA and the vector. The sgRNA sequences of a human papilloma virus 16 type gene suitable for CRISPR-Cas9 targeting editing and a human PD-1 gene are selected, and the plasmid vector of the sgRNA and a CRISPR-Cas9 nuclease gene expression vector are transferred to HPV16+ human cervical carcinoma cells and HPV16+ transplantation tumor-bearing mice, and can obviously inhibit HPV16 expression and inhibit tumor growth. The gene expression vector prepared in the invention has the advantages of simple method steps, good sgRNA targeting property, high CRISPR-Cas9 knockout efficiency, accurate targeting splicing of high risk HPV16 type and PD-1 genes, efficient reduction of the expression of the high risk HPV16 type gene, and obvious inhibition of the tumor growth through combined application.

The invention relates to a truncated human papillomavirus type 16 L1 protein, virus-like particles composed of the protein, a vaccine containing the virus-like particles, and an application thereof in preventing cervical cancer.

A fusion protein for inhibiting cervical cancer is disclosed, which comprises a peptide sequence of human papillomavirus type 16, a peptide translocating peptide for translocation, and a peptide within a carboxyl terminal fragment. The present invention further comprising a composition of antibody, which conjugates to E7 peptide, wherein the nucleotide sequence corresponding to the amino acid sequence of the E7 peptide is shown as SEQ. ID. NO.1.

The invention relates to a truncated L1 protein of the Human Papillomavirus Type 11, a virus-like particle consisting of the protein, a vaccine comprising said virus-like particle, and the use of the vaccine in the prevention of condyloma acuminatum or HPV infections.

The invention relates to a truncated L1 protein of the Human Papillomavirus Type 16, a virus-like particle consisting of the protein, a vaccine comprising said virus-like particle, and the use of the vaccine in the prevention of cervical cancer.

The invention discloses chimeric protein. The chimeric protein is obtained by inserting an epitope E7 of a human papillomavirus type 16 at a ring position of human papillomavirus type 16 (HPV 16) protein L1, namely between the 140th amino acid and the 141st amino acid, wherein the epitope E7 of the human papillomavirus type 16 has genes of protein (a) with the following codes: the protein is formed by an amino acid sequence shown by an MLDLQPETT epitope (12-20E7 for short), a RAHYNIVTF epitope(49-57E7 for short), an LLMGTLGIV epitope (82-90E7 for short) or a TLGIVCPI epitope (86-93E7 for short). The invention also discloses virus-like particles formed by aggregating the chimeric protein, a mixture formed by the virus-like particles and application of the virus-like particles and the mixture of the virus-like particles to the preparation of the vaccine for HPV 16 L1-virus-like particles.

The invention relates to a 6,11 type human papillomavirus fluorescence quantitative PCR detection method and kit thereof and PCR amplification primer and probe sequence for 6,11 type nucleotide segments of human papillomavirus. The primer sequence comprises: primer pair of human papillomavirus 6,11 type general upstream primer and general downstream primer, upstream primer complementation sequenceor downstream primer complementation sequence, also comprises primer sequences obtained in range of upstream primer extending towards 5'end by 10 basic groups and the upstream primer extending towards 3'end and 5'end by 10 basic groups. The probe sequence comprises: fluorescent probe: SEQ ID NO:3 and its complementation sequence, and probe sequence obtained in range of the extending SEQ ID NO:3 and its complementation sequence towards 3'end and 5'end by 10 basic groups. The method is used for quantitative detection of 6,11 type human papillomavirus with real clinical application value.

The invention provides a gene for encoding HPV (human papilloma virus) 6-type L2N120E7E6 fusion protein, expression vectors, a method, a bacteria strain and an application. The invention is characterized in that prokaryotic expression vectors are utilized to fuse HPV 6-type L2N-end 120 amino acids, E7 proteins and E6 proteins, and constructing the encoded gene according to the amino acid of the fusion protein. In the invention, the gene for encoding the HPV 6-type L2N120E7E6 fusion proteins is successfully designed and the expression vectors and the transformation bacteria strain of the high-expression HPV 6-type L2N120E7E6 fusion proteins are successfully constructed; and in the fusion proteins, the immunogenicity is strong, the immunoreaction of specific T cells can be induced, the tumor forming time of mice can be obviously delayed, and the growth of the tumors of 20% mice is completely restrained. The invention can be used for preventing and treating HPV 6-type infection and relevant diseases, such as genital wart.

The present invention relates to human papillomavirus type bivalent virus-like particle mixture, and human papillomavirus type bivalent virus-like particle mixing protein antigen and its construction method. The present invention includes one kind of cervical carcinoma virus HPV16 L1 antigen and one kind of condyloma acuminata virus HPV11 L1 antigen. The HPV11 L1 gene is first cloned from condyloma acuminata tissue through molecular geological method, and both HPV16 L1 gene cloned from cervical carcinoma tissue and the HPV11 L1 gene are then expressed in the insect cell expression system of baculovirus, so as to constitute recombinant virus strain Bacmid / HPV16-HPV11L1. Animal immunizing test shows that the constituted recombinant protein has excellent immunogenicity and immunoreactivity, and may be used as candidate antigen of gene engineering multivalent vaccine for preventing condyloma acuminata and cervical carcinoma.

The present invention is directed to a method of expressing the papillomavirus capsid protein coding sequence in a cell using an expression system under conditions facilitating expression of the protein in the cell.In another aspect of the invention, it has been discovered that virus-like particle(s) (VLPs), fragment(s), capsomer(s) or portion(s) thereof are formed from the papillomavirus capsid protein. It was further discovered that the virus-like particle(s) comprises antigenic characteristics similar to those of native infectious papillomavirus particles.In an embodiment of the invention, there is provided a method of expressing the L1 major capsid protein of human papillomavirus type-11 (HPV-11) in Sf-9 insect cells using the baculovirus expression system, and the production of HPV-11 virus-like particles.

The invention relates to a codon-optimized gene of a 16-type L2E7 recombinant protein of human papilloma virus (HPV), which can be expressed efficiently in colibacillus. The optimized gene can be expressed efficiently by being inserted in a prokaryotic expression vector pET9a, and the expression level accounts for 50% of all viruses; an expression protein is immune to mice after being purified. Proved by experiment results, the protein has the same immunogenicity with a protein which is not expressed by the optimized gene and can induce the release of specific gamma-1NF; proved by tumor growth inhibition animal experiments, protein immunity has evident inhibition effect on tumor growth and the growth of 90% of mouse tumor cells can be inhibited completely. The prokaryotic high-efficiency expression system of L2E7 recombinant protein, which is established by the optimized gene, can be used in the pilot scale production and drug discovery of preventive and curative vaccines of HPV16 infection and relevant cervical carcinoma.

Provided are an N-terminal truncated L1 protein of the Human Papillomavirus Type 58, a coding sequence and preparation method thereof, and a virus-like particle comprising the protein. Uses of the protein and the virus-like particle in the preparation of a pharmaceutical composition or a vaccine are further provided. The pharmaceutical composition or vaccine is used for prevention of HPV infection and a disease caused by HPV infection.

The invention relates to a truncated L1 protein of the Human Papillomavirus Type 18, a virus-like particle consisting of the protein, a vaccine comprising said virus-like particle, and the use of the vaccine in the prevention of cervical cancer.

The invention provides a polynucleotides gene fragment of recombinant human papilloma virus type 33 L1 protein, a carrier containing the gene fragment, a host cell containing the comprise, and a HPV33L1 protein pentamer expressed by the gene fragment and an anti-HPV type 33 infection vaccine composed of the pentamer.

The invention relates to a recombinant human papilloma virus 16L1 protein and its use, and provides a new polynucleotides gene fragment coding the recombinant HPV16L1 protein, a vector containing the gene fragment, a host cell containing the vector, an HPV16L1 protein pentamer expressed by the gene fragment in translation, and an anti-HPV16 infection cervical carcinoma vaccine composed of the pentamer.

Five optimized genes of capsid L1 protein from human papillomavirus type 16, 58, 18, 6 and 11, which are modified by using insect's preferred codons and so on. Method for modifying those genes to express more highly in insect cells. Virus-like particle's vaccine compositions comprising HPV L1 proteins or their functional relatives produced by using those modified genes. Those optimized genes of L1 can be used to produce HPV 16 VLP, HPV 58VLP, HPV 18VLP, HPV 6 VLP, HPV 11VLP in insect cells. Yields of virus-like particles derived from those optimized HPV L1 genes are high. Mixed multivalent vaccines comprising above optimized HPV L1 genes can be used to prevent and treat multiple HPV infection and diseases related with it.

The invention discloses an HPV 16 type E7 protein functional antagonist peptide and an encoding gene and an application thereof. The amino acid sequence of the HPV 16 type E7 protein functional antagonist peptide is shown in a sequence 1 in a sequence list. The HPV 16 type E7 protein is taken as the target protein, micromolecular short peptides which can be combined with the HPV 16 type E7 protein and is used for inhibiting the activity of combination between the HPV 16 type E7 protein and the pRb protein are screened to obtain a group of HPV 16 type E7 protein functional antagonist peptides with consistent sequences, and the HPV 16 type E7 protein functional antagonist peptides can be taken as the drug candidate for curing the cervical carcinoma induced by the HPV infection. The HPV 16 type E7 protein functional antagonist peptide has wide application value and broad market prospect in the preparation of the drug for curing the cervical carcinoma induced by the HPV infection.

Disclosed is a vaccine antigen capable of inducing a cross-reacting and neutralizing antibody directed against a high-risk-type human papillomavirus. Specifically disclosed are: a chimeric protein comprising an L2-epitope of a human papillomavirus (HPV) type-16 inserted in a loop region of a human papillomavirus type-16 L1 protein; and a capsid which is an aggregate of the chimeric protein. The loop region to which the L2-epitope is to be inserted is located between an amino acid residue at position-430 and an amino acid residue at position-433. The L2-epitope has an amino acid sequence represented by any one of the following formulae: YKTCKQAGTCPPDIIPKVEG (SEQ ID NO: 2) (18-32 L2-epitope); GGLGIGTGSGTGGRTGYIPL (SEQ ID NO: 3) (56-75 L2-epitope); and DPVGPLDPSIVSLVEESSFI (SEQ ID NO: 4) (96-115 L2-epitope).

The invention provides a novel polynucleotide gene segment of coded recombinant protein HPV6L1, a vector containing the gene segment, a host cell containing the vector, a protein HPV6L1 pentamer obtained by translating and expressing the gene segment, and an HPV6 infection-resistant condyloma acuminate vaccine consisting of the pentame.

The invention relates to a recombinant human papilloma virus 18L1 protein and its use, and provides a new polynucleotides gene fragment coding the recombinant HPV18L1 protein, a vector containing the gene fragment, a host cell containing the vector, an HPV18L1 protein pentamer expressed by the gene fragment in translation, and an anti-HPV18 infection condyloma acuminatum vaccine composed of the pentamer.

The invention discloses a method, primer, probe and kit for identifying various gene mutations or substance genetic typing. In one embodiment, the disease is pathogenic. In the other embodiment, the invention is applied to detection of multidrug-resistant Mycobacterium tuberculosis, hepatitis B virus, beta-globulin mutation, thrombophilia related mutation; or various sexually transmitted pathogens, consisting of Trichomonas vaginalis, Chlamydia trachomatis, Neisseria gonorrhoeae, Mycoplasma genitalium, Mycoplasma hominis, Ureaplasma urealyticum, Ureaplasma parvum, Treponema pallidum, Herpes simplex virus 1 , Herpes simplex virus 2, Human papillomavirus type 6, and Human papillomavirus type 1.

The invention relates to a method for guiding the expression of major capsid proteins L1 of HPV16 (human papilloma virus 16) in pichia pastoris by using a formaldehyde dehydrogenase promoter, wherein the expression of major capsid proteins L1 of HPV16 is controlled by a pichia pastoris formaldehyde dehydrogenase promoter. The method disclosed by the invention mainly comprises the following steps: i) building a pichia pastoris expression vector guided by a pichia pastoris formaldehyde dehydrogenase promoter; and ii) under the guidance of the formaldehyde dehydrogenase promoter, expressing the major capsid proteins L1 of HPV16 in different carbon / nitrogen source combined culture media. According to the invention, based on a pichia pastoris expression system, and starting with the regulating characteristics of the formaldehyde dehydrogenase promoter, the optimized expression of major capsid proteins L1 of HPV16 is implemented in the different carbon / nitrogen source combined culture media according to the characteristics of the promoter, so that the efficient expression of the major capsid proteins L1 of HPV16 is realized, thereby obtaining a low-cost virus-like particle preparation technology and implementing the low-cost preparation of human papilloma virus vaccines.

The present invention provides methods, primers, probes, and kits for identifying gene mutation or material genotyping. In one embodiment, the materials are disease-causing agents. In another embodiment, the present invention is applied to detecting multidrug-resistant mycobacterium tuberculosis, hepatitis B virus, beta-globulin mutation, thrombosis-related mutations; or multiple sexually transmitted diseases causing agents, including but not limited to, trichomonas vaginalis, chlamydia, neisseria gonorrhoeae, mycoplasma, mycoplasma hominis, ureaplasma urealyticum, Mycelia, Treponema pallidum, Herpes simplex virus type 1 and type 2 and human papilloma virus type 6 and type 11.

The invention provides an L1 major capsid protein gene of human papillomavirus type 16 and a pichia pastoris strain expressing the gene. The gene sequence of the L1 protein is the gene sequence which is optimized by codon preferred by pichia pastoris, and corrects the optimized codon frequency to express the L1 protein. The L1 major capsid protein gene of human papillomavirus type 16 is an optimized L1 gene, and has the advantages that the optimized gene is more suitable for high expressing target proteins in pichia pastoris host and can meet the requirement of industrial production; meanwhile, a pichia pastoris expression system is adopted, so the cost is low, the yield is high, and the product nature is more uniform and stable.

The invention discloses a proposal for constructing gland related virus containing 16 type antisense E7 gene of human papilomavirus(HPV), and particular application of a recombination gland related virus constructor obtained by the proposal in tumor treatment. By techniques of full length enlarging by the PCR method, enzyme cutting, connection, homologous recombination, transfection, purification of gland related virus and the like, the recombination gland related virus constructor is obtained. The technical characteristic comprises that a 16 type E7 gene cDNA segment of the full length exogenous HPV is oppositely inserted in a NotI enzyme cutting site. The recombination gland related virus constructor has the following obviously technical advantages: chromosomes can be integrated in a fixed site; the inserted reverse exogenous gene is a full length segment, and has good specificity; and the constructor has strong effect on cells infected with the 16 type HPV, and has prevention function. The recombination gland related virus constructor provides an ideal gene target point specificity treatment and prevention application way for treating and preventing tumor gene.

The present invention relates to a novel nucleic acid sequence (HPV16 L1S) encoding antigenic HPV16 L1 protein as provided in SEQ ID NO: 1, wherein the sequence has atleast one modified codon for optimum stability of recombinant plasmid vector when transformed in the prokaryotic micro-organism for improved immunogenicity of the resulting prokaryotic micro-organism. The invention further relates to constructing recombinant vectors pFS14nsdHPV16L1 and pFS14nsdHPV16 kan L1S harboring SEQ ID NO: 1, wherein the former carries Ampicillin and the latter, Kanamycin as a selection marker. The invention also relates to an attenuated strain of a prokaryotic micro-organism transformed with nucleic acid encoding HPV16 (Human Papillomavirus) major capsid protein and expressing the corresponding protein. In addition the invention discloses a process of producing a vaccine based on prokaryotic micro-organism for the treatment of papillomavirus infection and associated risk of cancer.

A polypeptide drug or bacterin for curing tumor caused by Human Papillomavirus Type 18 is characterized in that the aminophenol order is amido end- VAWDSVYYM- carboxyl end, which is a multi-copy serial structure with lysine as a joint and a plurality of aminophenol orders connected. The present invention has the advantages of convenient transport and storage as well as automatic mass production. The present invention also has the capacity of activating specific cytotoxic T lymphocytes (CTL) and the specific CTL cell can kill, damage and dissolve target cells with the relative tumor sign Human Papillomavirus Type 18. The present invention has obvious effect and no side effect.

The invention discloses a quadruple fluorescent quantitative PCR typing detection method for common types of human papillomavirus. It includes a four-fold PCR reaction system based on fluorescent quantitative PCR technology, including forward and reverse primers and AllGlo fluorescent probes for four human papillomavirus (HPV) genotypes, which can be used under suitable PCR conditions. Simultaneously detect the DNA load of HPV-6, -11, -16, and -18 in one reaction tube at most, including the design of specific primers and fluorescent probes, the construction of standard molecules, the preparation of standard curves, and the detection of clinical specimens. The present invention can easily and quickly detect clinically common four-type HPV infection simultaneously and quickly in one reaction tube, realize simultaneous typing and detection of four-type HPV by single-tube PCR, and can quantitatively detect, simple and fast operation, high sensitivity and specificity , good repeatability, accurate and reliable results, and has great clinical value for early prevention and treatment of genital warts and cervical cancer in women caused by common types of HPV, blocking the source of infection, reducing HPV infection, and monitoring clinical efficacy.

Disclosed is a HPV-16 peptide antigen, and the antigen comprises a peptide sequence of SEQ ID No: 1. The peptide antigen of the invention is able to bind with MHC class I antigen on the cell and be presented on the cell surface. It can be recognized by cytotoxic T lymphocytes (CTLs), therefore induces CTL responses for cytolysis to control the replication and expression of the HPV-16.