56 results about "Flagellin" patented technology

The present invention relates to a flagellin fusion protein and the use thereof, and, more specifically, to a fusion protein comprising a human IgG4 Fc variant and the use thereof using toll-like receptor 5 (TLR5) stimulation activity, wherein the human IgG4 Fc variant has mutation preventing a Fab-arm exchange.

The invention relates to the technical field of microbial metabolism engineering, and discloses a genetically engineered bacterium for high-yield L-histidine as well as a construction method and application of the genetically engineered bacterium. A nucleoside triphosphate pyrophosphate hydrolase coding gene mazG gene and a flagellin synthesis gene flhC gene are knocked out at the same time, an o-aminobenzoic acid phosphoribosyltransferase coding gene trpD gene and a ribulose phosphate 3-isomerase coding gene rpe gene are knocked down, and the recombinant protein is obtained. A ribose-5-phosphate isomerase gene coding rpiA gene and a lysine exponential coding gene lysE gene are overexpressed, and a promoter of 6-phosphoglucose isomerase pgi is replaced by a PfliC promoter, so that key substances for synthesis of L-histidine can be effectively accumulated, the fermentation yield of L-histidine is greatly increased, and compared with an initial strain, the yield of L-histidine is increased by 33.3%.

The invention discloses a salmonella typhimurium EN-VNP strain based on flagellin and LPS attenuation as well as a preparation method and application of the salmonella typhimurium EN-VNP strain, and particularly relates to the technical field of biology. The bacterial strain is formed by sequentially knocking out or silencing flagellin genes fliC and fljB, a phosphoethanolamine transferase gene eptA, a 4-amino-4-deoxy-L-arabinose transferase gene arnT and phosphotransferase yeiU on the basis of a salmonella VNP 20009 bacterial strain, and knocking in or overexpressing a phosphotransferase gene lpxE. The salmonella typhimurium EN-VNP strain disclosed by the invention has relatively high safety, the attenuated strain does not contain flagellin, and a lipid A molecule of LPS only retains one phosphate group, so that the safety of the strain is improved, and the strain is more suitable for serving as a vaccine and a drug delivery carrier.

The invention discloses a carrier protein with an adjuvant function and application thereof. The carrier protein with the adjuvant function has a sequence as shown in SEQ ID NO. 1. According to the carrier protein with the adjuvant function, a part of sequence is intercepted from the flagellin FliC of Burkholderia cepacia, so that the adjuvant activity of the flagellin is reserved, and the immunogenicity and antigenicity of the flagellin are reduced; when being applied to a fusion protein or a small-molecule drug, the fusion protein or the small-molecule drug can enhance the immunogenicity, so that the fusion protein or the small-molecule drug has important significance on subunit vaccines, polypeptide vaccines or small-molecule drug carriers.

Provided herein are recombinant microorganisms that express a subject polypeptide. Microorganisms can comprise an expression construct comprising a flagellin promoter operatively linked with a heterologous nucleotide sequence encoding the subject polypeptide. The flagellin promoter sequence can comprise a genetic modification that reduces CsrA inhibition of translation. Microorganisms also can comprise a genetic modification that reduces FlgM inhibition of SigD initiation of transcription. The target polypeptide can be an aldehyde dehydrogenase. Such microorganisms are useful in the treatment of alcohol hangover.

The present invention relates to the modification of an attenuated live strain of Salmonella enterica serovar Typhi, wherein its native surface-exposed polysaccharide and flagellin antigens can be converted to, or enhanced by, those of other strains of Salmonella, including Salmonella enterica serovar Paratyphi, Salmonella enterica serovar Typhimurium, and Salmonella enterica serovar Enteritidis. The present invention also relates to a modified strain of Salmonella enterica serovar Typhi suitable for use as a component of a vaccine against typhoid enteric fever and salmonellosis.

The present invention discloses a subunit vaccine against chicken infectious anemia virus, belonging to the technical field of animal disease vaccines. The present invention provides a subunit vaccine against chicken infectious anemia virus, and the active ingredient is a recombinant fusion protein of the B cell epitope of chicken infectious anemia virus VP1 protein and Salmonella typhimurium flagellin. The B cell epitope of chicken infectious anemia virus VP1 protein is obtained by connecting the B cell antigen epitopes contained in chicken infectious anemia virus VP1 protein through a linker peptide. The Salmonella typhimurium flagellin is obtained by deleting the hypervariable region of Salmonella typhimurium flagellin FliC. After the fusion expression protein of the present invention is administered to 1-day-old SPF chickens, it can induce the production of specific IgG antibodies, and after 28 days of immunization, the immunized chicken flock can resist the challenge of the virulent strain of chicken infectious anemia virus, and 70% of the challenged chicken flock is protected from disease. The present invention has great potential for clinical prevention and control of chicken infectious anemia virus infection.

The invention relates to lipid nanoparticles containing an adjuvant for mucosal administration and a preparation method of the lipid nanoparticles. The lipid nanoparticles comprise total lipid, an adjuvant and an active ingredient, wherein the total lipid is composed of cationic lipid C12-200, non-cationic phospholipid, cholesterol and a derivative thereof, and pegylated lipid; wherein the adjuvant is selected from at least one of AS03, MF59, monophosphoryl lipid A, miniaturized monophosphoryl lipid A, ophiopogonin D, saponin QS21, QS7, QS17, QS18, CpG oligonucleotide, 2 '3'-ring GMP-AMP, 3 '3'-ring GMP-AMP, alpha GC, Trehalose-6, 6-dibehenate, cholera toxin B subunit and flagellin.

The invention discloses a porcine rotavirus VP8 protein dominant B cell neutralizing epitope peptide, a multi-epitope fusion protein and application of the multi-epitope fusion protein. The FliCS.T-VP8 multi-epitope fusion protein prepared by the invention is a multi-epitope fusion protein which takes salmonella typhimurium flagellin FliCS.T as a molecular skeleton and integrates the dominant B cell neutralizing epitope of the VP8 protein of the porcine rotavirus. According to the design, immunodominance epitope screening, multi-epitope fusion protein construction and genetic engineering technologies are fused, and the immunogenicity and immune protection efficacy of the fusion protein are remarkably enhanced. The FliCS.T-VP8 multi-epitope fusion protein not only can effectively induce high-level VP8 protein specific antibody response and cellular immune response, but also shows good porcine rotavirus neutralizing activity. In addition, the fusion protein has relatively high safety and stability, and has the potential of being used as an efficient subunit vaccine for preventing and controlling porcine rotavirus infection in the pig industry.

The present invention relates to a Bacillus subtilis flagellin variant and a use thereof, and more specifically to a flagellin variant and a use thereof, the flagellin variant having improved storage stability and reduced immunogenicity compared to flagellin derived from wild-type Bacillus subtilis.

Provided herein is a peptide array comprising a plurality of flagellin peptides corresponding to highly conserved peptide regions. For example, the peptide array comprises a plurality of Lachnospiraceae flagellin peptides, which can be selected from a hinge region of Lachnospiraceae flagellin. Also provided is antibody of fragment thereof that binds to one or more of the plurality of the flagellin peptides in the peptide array. The peptide array and antibody are useful in determining an immunosignature from a biological sample from a subject. The immunosignature is useful in diagnosis of an immune-mediated disease, in monitoring progression of an immune-mediated disease, in identifying subjects susceptible to certain treatments, and in monitoring treatment response.

The invention belongs to the field of aquatic bacteria detection, and relates to a fluorescent array sensor as well as a preparation method and application thereof. The fluorescent array sensor comprises a plurality of DNA aptamers, the DNA aptamers respectively target lipopolysaccharide, peptidoglycan and flagellin, the DNA aptamers of the target lipopolysaccharide are EA5 and LA27, the DNA aptamer of the target peptidoglycan is Antibac2, and the DNA aptamer of the target flagellin is Aptamer 3; the sequence of the EA5 is shown as SEQ ID NO.1, the sequence of the LA27 is shown as SEQ ID NO.2, the sequence of the Antibac2 is shown as SEQ ID NO.3, and the sequence of the Aptamer 3 is shown as SEQ ID NO.4. The invention further discloses a kit for detecting the content of the EA5, the LA27, the Antibac2 and the Aptamer. According to the present invention, the six bacteria can be simultaneously detected, the detection limit is low, the operation process is simple, and the detection accuracy can achieve 97.6%.

The present invention relates to a DNA construct comprising a gene encoding a flagellin protein and a gene encoding an adjuvant protein. The present invention is an effective cancer treatment that selectively kills only cancer cells. The attenuated Salmonella strain is designed to produce an immunostimulatory substance in cancer tissue, thereby inducing a strong anti-cancer immune response. This can be useful as a preventive or therapeutic composition that significantly suppresses tumor size in primary cancers as well as metastatic cancers, thereby improving survival rates.

The present invention relates to a combination of (i) a flagellin polypeptide and (ii) an antibiotic, for the simultaneous or sequential use in the treatment of bacterial infections due to Pseudomonas aeruginosa antibiotic-resistant strain. The present invention also provides a flagellin polypeptide, for use in a method for enhancing sensitivity to an antibiotic of a patient suffering from bacterial infection, due to Pseudomonas aeruginosa antibiotic-resistant strain. Indeed, using an in vivo mouse model, inventors show that while standalone prophylactic flagellin attenuates infection caused by a MDR strain of PA (PAMDR), its combination with an antibiotic such as gentamicin (GNT) leads to a strong decrease in bacterial load in the lung and a significant reduction in cell infiltration and inflammatory cytokines. Moreover, mice receiving flagellin in combination with GNT showed a 100% survival rate, a result that GNT alone was unable to provide against PAMDR.

The present invention relates to a strain for controlling tomato bacterial wilt and promoting tomato growth, and to a method for controlling tomato bacterial wilt and a method for promoting tomato growth using same. A microbial agent containing the motile mutant strain or a metabolite of production change due to deletion of a flagellin protein gene, according to the present invention, can control tomato bacterial wilt at a level comparable to registered levels of synthetic pesticides by an eco-friendly and low-input method. In addition, a microbial formulation comprising a non-pathogenic Ralstonia solanacearum Gj707 strain or RSp1180 protein, which is the adhesin protein represented by SEQ ID NO: 2, according to the present invention, can control tomato bacterial wilt at a level comparable to registered levels of synthetic pesticides. Furthermore, the tomato root adhesion protein of the bacterial wilt pathogen can be utilized for developing agents with site-specific action against bacterial wilt, thereby enabling efficient and health-friendly management of bacterial wilt, for which no dedicated agents are currently registered. Unlike general synthetic pesticides, such formulations also promote tomato growth, and thus are expected to increase the production and income of tomatoes in farmhouses.

The invention relates to the technical field of molecular biology, and particularly discloses a preparation method and application of universal influenza virus nasal spray subunit vaccine candidate protein, and the preparation method comprises the following preparation steps: synthesis of a recombinant plasmid pET-28a-FluA; the preparation method comprises the following steps: carrying out transformation induction on a recombinant plasmid pET-28a-FluA to prepare a recombinant protein; purifying the recombinant protein to obtain a target protein; preparing a chitosan solution, and emulsifying the chitosan solution and a target protein to obtain a universal influenza virus nasal spray subunit vaccine candidate protein; according to the invention, a plurality of epitopes with good immunogenicity and conservative property are connected in series through bioinformatics prediction to construct the recombinant protein, and meanwhile, the protective capability and the protective range of the recombinant protein are increased to the greatest extent by utilizing the enhancement effect of an intramolecular adjuvant flagellin on mucosal immunity; compared with a traditional subunit vaccine, the subunit vaccine has better immunogenicity and a wider protection effect.

The invention belongs to the technical field of plant disease control, and discloses flagellin and application thereof in a plant immune resistance inducer. The flagellin is separated and extracted from bacillus amyloliquefaciens Ba168, has better bacteriostatic activity and has an immune induction effect on tobacco. According to the invention, a gene engineering technology is utilized to realize mass expression and purification of the flagellin, and it is identified that the flagellin can induce plant immune response, plant cell necrosis, plant reactive oxygen bursting and increase of plant defensive enzyme activity. The flagellin can induce the resistance of plants to PVX, and a novel resistance inducing material is provided for prevention and treatment of PVX.

This application relates to compositions and methods used to treat animals with at least one microbial protein, such as a flagellin protein. In embodiments, application relates to compositions and methods to treat animals with a fusion protein comprising at least one adjuvant protein, such as a flagellin protein. In embodiments, the composition further includes at least one adjuvantic protein fused with at least one antigen protein, and the fusion protein is expressed in diatoms.

The application belongs to the technical field of biology, and discloses a preparation method of a Salmonella typhimurium flagellin and Senecavirus antigen fusion protein. The application selects Escherichia coli Rosetta (DE3) as a host, fuses a Salmonella typhimurium flagellin gene and a Senecavirus antigen gene, optimizes codons, and then clones the fusion protein into a pET-28a (+) vector, so that the obtained fusion protein can keep the adjuvant activity of the flagellin, improve the solubility and expression level of VP2, and the yield can reach 40 mg / L, thereby greatly reducing the expression and purification process cost. The fusion protein provided by the application can activate the TLR5 path and activate mouse macrophages, and the production process is simple and low in cost, so that the application has important significance for the research and production of Senecavirus subunit vaccines.

The present invention relates to the manufacture and application of flagellin immunopotentiating derivatives with TLR5 activity using an eukaryotic cell expression system. The flagellin derivative according to the present invention has a high immunostimulatory effect and can be used in various vaccine and immunotherapy compositions.

The present invention relates to a DNA construct comprising: a gene encoding flagellin and a gene encoding an immunopotentiator (or adjuvant) protein. In order to achieve effective cancer treatment by selectively killing only cancer cells, the attenuated Salmonella strains of the present disclosure are designed to produce immunogenic substances in cancer tissue to induce a strong anti-cancer immune response to significantly inhibit tumor size in primary and metastatic cancers. Therefore, the strain can be advantageously used in a prophylactic or therapeutic composition for increasing survival rate.

The present disclosure relates to IL-13 / IL-4 / Flagellin fusion proteins for the stimulation of the production of anti-IL-4 and anti-IL13 antibodies in a non-human mammal, in particular antibodies that reduce Interleukin (IL)-4 and Interleukin (IL)-13 in a subject.

The invention relates to application of recombinant flagellin in preparation of an anti-Vibrio harveyi medicine for aquatic animals. Belongs to the technical field of bioengineering and immunology. The amino acid sequence of the recombinant flagellin is shown as SEQ ID No. 2; the nucleotide sequence of the gene of the recombinant flagellin is as shown in SEQ ID No. 1; the recombinant vector comprises an expression vector and a gene of the recombinant flagellin; the expression vector is a prokaryotic expression vector pET32a; the recombinant bacteria comprise host bacteria and the recombinant vector; and the host bacterium is a Rosetta engineering strain. After the vaccine prepared from the recombinant flagellin is injected into a fish body for 4 weeks and 8 weeks, the immune protection rates on vibrio harveyi are 61.12% and 47.83% respectively; after the vaccine prepared from the recombinant flagellin is injected into a fish body, the expression of a specific antibody can be remarkably improved; after the vaccine prepared from the recombinant flagellin is injected into a fish body, the enzyme activity of serum can be remarkably improved, and innate immune response and inflammation and cellular immune response are activated.

The application belongs to the technical field of biological medicine, and discloses a recombinant protein subunit vaccine constructed by combining Salmonella flagellin and a COE region of porcine epidemic diarrhea virus (PEDV). The application specifically discloses a fusion protein, wherein the fusion protein comprises Salmonella flagellin FliC L3A a mutant and a PEDV structural protein connected to the C terminal of the Salmonella flagellin FliC L3A The subunit vaccine obtained by emulsifying the fusion protein and the adjuvant can effectively prevent the G2b subfamily of PEDV, initiate a downstream immune response, induce a series of immune responses of the body, make the mucosal immune system of the body produce a large number of PEDV-specific antibodies to resist the invasion of external pathogens, and has the effect of specific immune protection.

Provided are delivery immunostimulatory bacteria that have enhanced colonization of tumors, the tumor microenvironment and / or tumor-resident immune cells, and enhanced anti-tumor activity. The immunostimulatory bacteria are modified by deletion of genes encoding the flagella, or by modification of the genes so that functional flagella are not produced, and / or are modified by deletion of pagP or modification of pagP to produce inactive PagP product. As a result, the immunostimulatory bacteria are flagellin and / or pagP−. The immunostimulatory bacteria optionally have additional genomic modifications so that the bacteria are adenosine or purine auxotrophs. The bacteria optionally are one or more of asd−, purI−, and msbB−. The immunostimulatory bacteria, such as Salmonella species, are modified to encode immunostimulatory proteins that confer anti-tumor activity in the tumor microenvironment, and / or are modified so that the bacteria preferentially infect immune cells in the tumor microenvironment, or tumor-resident immune cells, and / or are modified to induce less cell death in immune cells than in other cells. Also provided are methods of inhibiting the growth or reducing the volume of a solid tumor by administering the immunostimulatory bacteria.

The invention belongs to the technical field of biology, and particularly relates to a flagellin mutant capable of enhancing TLR5 ligand activity and application of the flagellin mutant. An amino acid mutation site of salmonella typhimurium flagellin is selected, recombinant protein is expressed by using an escherichia coli system through a site-specific mutagenesis technology, in-vitro experiments prove that the recombinant protein shows good biological activity, and compared with a wild flagellin mutant, the recombinant protein can better activate the activity of a TLR5 ligand and can be used for preparing a TLR5 mutant. And a good foundation is provided for enhancing the activity of flagellin adjuvants and vaccines.

The present invention relates to a chimeric recombinant protein having a therapeutic effect on cervical cancer by fusing genetic modified E6 and E7, which are carcinogenesis-inducing proteins of human papillomavirus high-risk group type 16, with a fusion protein for increasing immunogenicity, the HPV type 16 E6, E7 chimeric recombinant protein fused with the flagellin fusion protein of the present invention showed the lowest tumor cell volume, and the immune response of specific T cells according to the recombinant antigen was significantly confirmed, and when the prophylactic effect was measured, it was confirmed that the volume of tumor cells was low and the antibody titer was increased, therefore human papillomavirus recombinant antigen of the present invention shows tumor treatment and prophylaxis and can be applied as a therapeutic / prophylactic vaccine composition.

The present invention relates to a FLT3L–flagellin hybrid adjuvant with enhanced antigen cross-presentation efficacy and a vaccine composition comprising same. The hybrid adjuvant of the present invention demonstrated significant therapeutic efficacy as an adjuvant for a therapeutic cancer vaccine in a preclinical mouse model of cervical cancer, resulting in complete tumor regression and sustained protection. In addition, the hybrid adjuvant of the present invention induced tumor-specific antigen CD8+ T cell responses and progenitor-exhausted CD8+ T cells (Tpex) due to an increase in cross-presentation by conventional type 1 dendritic cells (cDC1) in tumor-draining lymph nodes and the tumor microenvironment, and the combination of TCV having the hybrid adjuvant applied thereto and anti-PD-1 therapy significantly improves survival outcomes in anti-PD-1–resistant tumors, and thus can be advantageously applied in anticancer immunotherapy using therapeutic cancer vaccines.

The application belongs to the technical field of biology, and discloses a Salmonella typhimurium flagellin derivative and Senecavirus antigen fusion protein and application thereof. The Salmonella typhimurium flagellin derivative gene and the Senecavirus antigen gene are fused, and after codon optimization, the fusion protein is cloned into a pET-28a(+) vector, and the fusion protein is expressed by using an Escherichia coli system. In-vivo experiment verification shows that the fusion protein has good biological activity, can activate TLR5 ligand activity, and promote macrophage activation, and lays a foundation for development of a Senecavirus subunit vaccine.

The invention belongs to the technical field of biology, and particularly relates to salmonella typhimurium flagellin capable of enhancing human TLR5 binding affinity and application of the salmonella typhimurium flagellin. A key amino acid mutation site of flagellin is screened and determined, and recombinant expression is realized in an escherichia coli system by adopting a site-directed mutagenesis technology. In-vitro experiments prove that the derivative can significantly activate the function of a TLR5 ligand. The invention solves the problem that the existing flagellin has a limited effect on activating the activity of the human TLR5, and provides a new technical scheme and theoretical basis for the application of the flagellin in the fields of vaccines, immunotherapeutic agents and radioprotectants.