74 results about "HBcAg" patented technology

The invention relates to the technical field of gene engineering, in particular to an immune enhanced gene vaccine of a hepatitis B virus core antigen (HBcAg). The vaccine is an immune stimulating complex (ISCOM) vaccine consisting of HBcAg and OX40 ligand (OX40L) dual-gene co-expression recombinant eukaryotic vector, saponin Quil A, cholesterol and lecithin. The vaccine can promote immune response of specific T cells of the HBcAg and efficiently induce the anti-virus replication capacity of immune cells, and has good application prospect in the field of hepatitis B virus infection resistance. The invention also relates to a preparation method for the vaccine with simple and convenient operation and low cost.

The invention discloses a novel coronavirus vaccine based on chimeric virus-like particles, and the vaccine contains chimeric virus-like particles as effective ingredients. The chimeric virus-like particles are mainly formed by the aggregation of a type of recombinant chimeric proteins, and the recombinant chimeric proteins are obtained by fusing the S protein receptor binding region of SARS-CoV-2virus into the major immune determining region of HBcAg. The vaccine of the present invention can produce a strong immune response in human bodies, and humans can resist new coronavirus infections after immunization.

The present invention is a composition that comprises at least two hepatitis B virus surface antigens (HBsAgs), fragments thereof and/or nucleic acids encoding them, the HBsAgs differing in HBV genotype in the S region and/or pre-S1 region and the composition containing no HBV core antigen (HBcAg) or nucleic acid encoding that antigen. The present invention also includes pharmaceutical compositions, especially vaccines, comprising these compositions for the prevention and/or treatment of an HBV infection or an HBV-mediated disease. The present invention further includes a method of preparing a patient-specific medicament for the therapeutic treatment of hepatitis B.

The invention relates to the field of molecular biology and immunology, in particular to an application of an HBcAg (hepatitis B core antigen) virus-like particle serving as a cervical cancer therapeutic vaccine carrier. A preparation method comprises steps as follows: an HPV16 E749-57CTLs epitope peptide fragment is selected, a DNA (deoxyribose nucleic acid ) fragment of the HPV16 E749-57CTLs epitope peptide fragment is inserted between 78 and 79 amino acids of the HBcAg through genetic recombination, an obtained recombinant plasmid pHBcAg-E749-57 is converted into Escherichia coli DH5alpha, and an HBcAg virus-like particle vaccine presenting E749-57 is obtained after induction expression and purification. After a tumor-bearing mouse is immunized with the virus-like particle vaccine, the body of the mouse can be induced to generate a higher HPV16E7 specific cellular immunologic response, and growth of tumors is remarkably inhibited.

The hepatitis B virus (HBV) capsid is made up of a single species of protein called the core antigen (HBcAg) which self-assembles into particles. The particles are highly immunogenic and are able to present heterologous epitopes to the immune system when the epitopes are inserted into a surface-exposed region of the particles called the “e1 loop”. The structural building blocks of the particles are tightly associated dimers of HBcAg in which the adjacent e1 loops are closely juxtaposed. It is proposed that sequences inserted into the e1 loop are conformationally restrained in the assembled particles when presented in monomeric core protein. The invention seeks to solve this problem by covalently linking core proteins as tandem copies (e.g., as dimers) so that insertions can be made independently in each copy. This is particularly useful for insertion of large sequences into the e1 loop because it allows such sequences to be inserted into just one copy of the core protein per tandem repeat, thereby reducing potential conformational clashes in assembly. Alternatively, a different sequence may be inserted into each e1 loop of a tandem repeat, thus increasing the flexibility of HBcAg particles as an epitope delivery system.

A therapeutic DC vaccine for preventing and treating chronic hepatitis B is a hepatitis B specific DC vaccine carried by both HBsAg and HBcAg. The dendritic cells coming from mononuclear cells are carried by both recombinant HBV surface antigen and recombinant HBV core antigen.

The invention closes HBV core region through adopting siRNA technique to combine with a nanometer technique and inhibits the expression of HBcAg, and finally inhibits the expression and copy of HBV. The invention has wide target and obvious effect, which improves gene inducing efficiency through adopting the nanometer technique.

The invention provides a protein comprising a first and a second copy of hepatitis B core antigen (HBcAg) in tandem, in which one or both of the copies of HBcAg comprises influenza virus A surface polypeptide M2 or a fragment thereof in the e1 loop.

The invention discloses an immune-enhanced hepatitis B therapeutic multivalent vaccine and a preparation method thereof. The vaccine is a compound of an HBcAg epitope peptide, an HBsAg epitope peptide, an HBeAg epitope peptide and an LIGHT gene recombinant expression vector, wherein the HBcAg epitope peptide, the HBsAg epitope peptide and the HBeAg epitope peptide respectively comprise epitopes HBcAg87-95/HBsAg172-180/HBeAg147-155, a transmembrane sequence of HIV-Tat49-57, an endoplasmic reticulum retention signal sequence KDEL and a connector sequence, the transmembrane sequence is arranged at the amino terminal, the endoplasmic reticulum retention signal sequence is arranged at the carboxyl terminal, the epitopes are connected with the transmembrane sequence or the endoplasmic reticulum retention signal sequence through the connector sequence, and the vaccine can successfully enter cells and target the endoplasmic reticulum and effectively stimulate hepatitis B virus protein epitope to enter an MHC-I antigen presentation pathway to activate CTL (Cytotoxic Lymphocyte) responses.

The invention discloses a hepatitis B virus dendritic cell therapeutic vaccine and a preparation method thereof. The preparation method thereof includes the steps that: HBcAg-Fc digenic co-expression recombinant eukaryotic vector is constructed; the obtained digenic co-expression recombinant eukaryotic vector is added with saponin Quil A and then is added with cholesterol and lecithin, ice-bath ultrasonic processing is carried out, so that solution is fully mixed and emulsified, PBS dialysis is carried out, and the obtained micro floccus is immunostimulating complex; and the obtained immunostimulating complex activates and simulates dendritic cell, thus obtaining the cell therapeutic vaccine. The vaccine carries out high efficiency transfection on dendritic cell, the generated fusion protein targets the dendritic cell in high efficiency by virtue of Fc segment, thus activating the dendritic cell, promoting immune response of hepatitis B virus core antigen specificity T cell and inducing the antivirus replication capability of immune cell in high efficiency, so that the vaccine can be taken as hepatitis B virus therapeutic vaccine.

The invention discloses an EGFR and HER2 combined polypeptide epitope vaccine; two B cell epitope mimic polypeptide I and II of HER2 of the HER family, and a B cell epitope mimic polypeptide of EGFR of the HER family are inserted in a serial form between the 78th amino acid and the 79th amino acid of a hepatitis B core antigen HBcAg so as to form fusion protein which is the combined polypeptide epitope vaccine. The EGFR and HER2 combined polypeptide epitope vaccine of the invention breaks through the limitations of existing single epitope vaccines, and the combined epitope vaccine is providedwith more extensive antineoplastic activity.

The invention relates to a hepatitis B virus multi-epitope fusion protein and a preparation method and application thereof. The fusion protein is obtained by inserting hepatitis B virus multi-epitope fusion peptide (with the sequence shown as SEQIDNo.1) formed by serially connecting HBsAg313-321, HBsAg335-343, Pol150-159, Pol455-463 and Padre epitopes through connecting peptide between amino acidat the 78th position and amino acid at the 79th position of a hepatitis B virus core protein; and the preparation method comprises the following steps of: constructing a hepatitis B virus multi-epitope fusion protein expression plasmid pET28-HBc-HP; performing isopropyl thiogalactoside (IPTG) inducing expression by using an Escherichia coli expression system; and purifying by using affinity chromatography. The fusion protein carries a plurality of supertype epitopes of hepatitis B surface antigen (HBsAg), hepatitis B core antigen (HBcAg) and polymerase, is viral particles, has the advantages of strong immunogenicity, wide applicable range and the like, and can be used for preparing therapeutic hepatitis B vaccines.

The invention relates to the technical field of biomedicines. The currently reported hepatitis B virus vaccine mainly comprises one or several of HBsAg, preS 1, preS2 and HBcAg, some cell factor genes with immunological enhancement function or lymphocyte epitope genes and the like, however, the immunoprophylaxis effect and treatment effect of the vaccine are always unsatisfactory. The invention provides a hepatitis B vaccine which comprises hepatitis B virus surface antigen gene HBsAg, core protein gene HBc and e-antigen gene, also comprises a human microRNA181a precursor gene sequence, and can assist stimulating the body immunity pathway. The construction process of the vaccine relates to PCR, enzyme cutting, connection, conversion and other molecularly biological operating means. The hepatitis B vaccine has the advantages of effectively activating the body to produce a specific antibody against the hepatitis B virus, stimulating body cell immunity and secreting various cell factors. Therefore, the aim of treating chronic hepatitis B is fulfilled.

The invention provides a protein comprising a first and a second copy of hepatitis B core antigen (HBcAg) in tandem, in which one or both of the copies of HBcAg comprises a single-domain antibody fragment in the el loop.

The invention relates to the technical field of genetic engineering and biomedicines, in particular to an antigen epitope peptide and application thereof. The amino acid sequence of the antigen epitope peptide provided by the invention is shown as SEQ ID NO:1; the antigen epitope peptide is obtained by connecting HBcAg (or/and HBsAg) and surface antigen epitope peptide HBV Pre-S2 in series. Experimental results show that the mature of dendritic cells can be effectively induced, the antigen presentation capacity can be effectively improved and more cell factors can be secreted by adopting the antigen epitope peptide provided by the invention; after the antigen epitope peptide is co-cultured with lymphocytes, specific cytotoxicity can be produced to kill cells, so that the tumour killing efficiency is improved; the growth of a tumour can be effectively inhibited; an anti-tumour effect is achieved; a new choice is provided for clinical treatment of hepatitis B and related liver cancer thereof.

The invention relates to a recombinant fusion protein HSP65-HbcAg, which is fused from heat shock protein 65 and multi-epitope hepatitis B virus HbcAg, in the recombinant fusion protein, the multi-epitope hepatitis B virus HbcAg comprises 5 different antigen epitopes through interconnection by mathematical combination modes. the recombinant fusion protein can make hepatitis B virus HbcAg specific cell toxic T lymphocyte CTL eradicate the cells infected by hepatitis B virus, thus achieving the goal of treating and/or preventing hepatitis B. The invention also provides the nucleic acid sequence encoding the recombination fusion protein.

The present invention is related to the branch of medicine, particularly to the new formulations of vaccine antigens.

The technical objective pursued with the present invention is, precisely, the development of formulations that are able to enhance the immune response to mucosally administered antigens, minimising the number of compounds in the formulation and generating strong mucosal and systemic responses through a synergic interaction between the antigens in the formulation.

These formulations enable: a) to broaden the spectrum of the anti-hepatitis B immune response, containing as main compounds HBsAg and HBcAg, b) to enhance the response against HBsAg with a viral nucleocapsid c) to generate combined vaccines through the mucosal route with HBsAg as a central antigen. Stabilizers and preservatives can be introduced.

The formulations of this invention can be applied in the pharmaceutical industry as human or veterinary vaccine formulations.

The present invention relates to a composition, comprising: i) an HBcAg<1-x>, wherein x is a whole number from the range from 100 to 160, a fragment of this antigen, a variant of this antigen or of the fragment of this antigen or at least two thereof, ii) an adjuvant comprising a saponin or a saponin derivative or a mixture of at least two thereof, and optionally iii) an HBsAg, a fragment from the antigen, a variant of this antigen or of the fragment of this antigen or at least two thereof. The invention also relates to a process for production of a composition, the composition obtainable by this process, a pharmaceutical formulation, the use of the composition or of the pharmaceutical formulation for treatment and/or prevention of HBV infections and HBV-mediated diseases, the use of the composition or of the pharmaceutical formulation for production of a pharmaceutical for treatment and/or for prevention of HBV-infections and HBV-mediated diseases as well as a process for treatment and/or for prevention of HBV-infections and HBV-mediated diseases.

The invention provides a fusion protein related to HBV, a preparing method thereof and an application thereof. Particularly, a prokaryotic expression carrier related to HbcAg is built and expressed through the genetic engineering technology, and the obtained fusion protein can remarkably induce an HBV-specificity CTL reaction in vitro and vivo.

The invention provides a purification method for HBcAg (hepatitis B virus core antigen)-VLP or an HBcAg-VLP derivative. The method comprises the following steps: treating a loading solution containingHBcAg-VLP and a loading solution containing the HBcAg-VLP derivative by hydrophobic interaction chromatographic packing, and purified HBcAg or the purified HBcAg-VLP derivative is obtained. Accordingto the purification method, the problems of numerous steps, low yield, expensive consumables and the like of existing methods are solved, purification can be achieved by only one-step hydrophobic chromatography; when a gel filtration refining method is not used for treatment, the yield can reach up to 90% or higher, purity can reach up to about 86%, and the purification effect is greatly improved; if the purification method is supplemented with refining and purification methods such as further gel filtration, the purity can reach 99% or higher. The purification method has good application prospect and very high application value.

The invention relates to an antigen epitope, in particular to an immune dominant HLA-A3 (human leukocyte antigen A3) super-type restrictive CTL (cytotoxic T lymphocyte) epitope of a hepatitis B virus core antigen (HBcAg) and an identification method for the super-type CTL epitope. The super-type CTL epitope consists of the following amino acid sequence: Leu-Leu-Asp-Thr-Ala-Ser-Ala-Leu-Tyr-Arg. The method is also suitable for identification of other antigen super-type CTL epitopes, and can provide a useful tool for design and research of therapeutic polypeptide vaccines. The invention also relates to application of the super-type CTL epitope in preparing hepatitis B therapeutic polypeptide vaccines, and provides a new strategy and a new method for efficient research of the hepatitis B therapeutic polypeptide vaccines. The method is expected to break through hepatitis B immune tolerance, reconstruct the immune function of cells and efficiently inhibit and clear hepatitis B viruses.

The invention relates to the field of biomedicine, in particular to a preparation method of a subunit vaccine based on prokaryotic expression of a norovirus antigenic epitope. The invention provides the prokaryotic expressed subunit vaccine which is used for stimulating a body to produce a specific antibody against norovirus, and is characterized in that the norovirus recombinant antigen sequenceis as shown in SEQ ID NO: 1, source: NCBI Reference Sequence: NC_029646.1, a neutralizing epitope of human norovirus GII type: 105 bp, and an expression vector: a modified hepatitis B core antigen (HBcAg) gene cloned into a pThioHisA expression vector.

The invention provides an immunity-enhanced type virus-like particle for presenting recombinant cat sensitinogen rFel d 1 protein, an expression vector of the immunity-enhanced type virus-like particle and preparation and application of the immunity-enhanced type virus-like particle. The protein subunit forming the virus-like particle is fused protein of hepatitis B core antigen and polypeptides with the amino acid sequence of SEQ ID NO:1. The amino acid sequence of the fused protein is formed by inserting the sequence SEQ ID NO:1 of polypeptides between the 78th to 81st amino acid of hepatitis B core antigen and replacing the 79th to 80th amino acid of hepatitis B core antigen. By means of HBcAg-fFel d 1 fused protein, the immunogenicity of fFel d 1 can be remarkably improved, and therefore the detection sensitivity of fFel d 1 recombined and expressed in vitro in application of in-vitro diagnosis cat allergies is improved.

By adopting methods of fluorescent quantitation PCR, fluorescent quantitation RT-PCR, HBV DNA quantitation, HBsAg quantitation, HBeAg quantitation, cccDNA quantitation, Northern blot, Southern blot, Western blot, immunohistochemistry, and the like in the research and using the maintenance dose within the treating dose range for a long time (30-60 days), supernatant HBV DNA HBsAg HBeAg cultured by HepG-2.2.15 cells can completely disappear, HBsAg, HBeAg, HBcAg, and the like in the cells are completely turned to be negative, HBV DNA is in a high inhibited state, HBV cccDNA is completely negative, fluorescent quantitation RT-PCR detection finds that the mRNA for expressing HBsAg and HBcAg antigens in the cells is completely negative, and in addition, the curative effect of the atabrine is 30 times that of lamivudine as a first-line drug for resisting HBV in current clinic. In order to illustrate the action mechanism of the atabrine and the pyronaridine as a drug belonging to the same kind with the atabrine, an HBV genome is divided into 3 segments which are respectively inserted into an Xb1 position on a luciferase report carrier PGL3, a multifunctional microplate reader detects and finds that the light production value of the luciferase is remarkably reduced, which shows that the molecules of the atabrine and the pyronaridine as a drug belonging to the same kind with the atabrine can be nonspecifically combined with the HBV DNA in the cells, thereby inhibiting the copying of the virus and ensuring that the HBV DNA content of the virus in cells copied by the filial generation is reduced till to disappear. The patent requires protecting the application of 3 linked benzyl structures (named as ethyleneimine) and radicals such as CH3O-,-NH-, CL-, and the like for resisting HBV virus in clinic.

The invention relates to a method for preparing a chimeric monoclonal antibody capable of neutralizing EV71 (enterovirus 71). The chimeric monoclonal antibody for neutralizing EV71 or a fragment of the chimeric monoclonal antibody is characterized in that a heavy chain variable region of the chimeric monoclonal antibody or the fragment of the chimeric monoclonal antibody has an amino acid sequence shown in SEQ ID NO:1; a light chain variable region has an amino acid sequence shown in SEQ ID NO:2. EV71 VP4 (viral protein 4) protein is fused into an HBcAg (hepatitis B core antigen) virus-like particle to be prepared into fusion protein, and the fusion protein is taken as an antigen to endow a Balb/c mouse with immunity. A phage display antibody library technique is used for screening to obtain the monoclonal antibody capable of specifically neutralizing EV71 VP4 protein. The invention provides the method for preparing a potential monoclonal antibody medicine with broad spectrum neutralization activity and for preventing and treating a hand foot mouth disease.

The invention discloses a preparation method and an application of a human heat shock protein gp96. The method is characterized in that the human heat shock protein gp96 is expressed in an insect cell. The human heat shock protein gp96 expressed by an insect cell expression system does not bind with an antigen polypeptide, so the human heat shock protein gp96 can be well bound with HBcAg and HBsAg, thereby the immune functions of the human heat shock protein gp96 are enhanced, and HBV therapeutic vaccines developed with the human heat shock protein gp96 secreted and expressed by the insect cell as an adjuvant can greatly improve the anti-HBV curative effect of the HBV therapeutic vaccines.