33 results about "Flagellin" patented technology

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 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.

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 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.

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 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 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.

The invention relates to a novel Toll-like receptor 5 agonist as well as a preparation method and application thereof, and provides a flagellin variant of which the amino acid sequence is shown as SEQ ID NO: 1. Or a sequence having at least 80%, or at least 90%, or at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or at least 99.5%, or at least 99.9% identity with the amino acid sequence as shown in SEQ ID NO: 1. The flagellin variant not only has low immunogenicity, but also can effectively prevent extremely severe bone marrow acute radiation injury, is good in stability, and shows a potential application value in radiation injury protection.

The invention relates to bacillus flagellin FlgE serving as a specific ligand of animal intestinal tracts TLR5 and application of the bacillus flagellin FlgE in replacement of feed antibiotics, and belongs to the technical field of microbial engineering, FlgE interacts with the animal intestinal tracts TLR5 to promote colonization of probiotics in the animal intestinal tracts and competitively inhibit colonization and growth of harmful bacteria in the animal intestinal tracts, the animal is a pig or a chicken. According to the invention, in-vitro interaction between bacillus flagellin FlgE and pig source TLR5 as well as between bacillus flagellin FlgE and chicken source TLR5 is verified, and a novel TLR5 specific ligand protein is found, so that a novel thought and a novel direction are provided for researching a colonization mechanism of probiotic bacillus in intestinal tracts; related test results also provide theoretical and practical basis for development of safer and more efficient animal feed antibiotic substitutes based on probiotic bacillus.

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.

This invention discloses a sodium caseinate-based Pickering emulsion, its preparation method, and its application, belonging to the field of vaccine adjuvant technology. This invention is the first to propose using Salmonella Typhimurium flagellin and sodium caseinate as raw materials to formulate a Pickering emulsion. The prepared Pickering emulsion has the characteristics of small particle size, good biocompatibility, and high stability, and can induce a high immune response in the body. This invention is also the first to propose the application of the Pickering emulsion formulated using Salmonella Typhimurium flagellin and sodium caseinate as raw materials as a vaccine adjuvant. The Pickering emulsion prepared by this invention has a simple preparation process, requiring only mixing and homogenization, and can effectively load the model antigen OVA. This emulsion has good cellular safety and can promote the endocytosis of exogenous antigens by macrophages. The emulsion can induce the body to produce high levels of antibody titers, with antibody titers reaching more than 120,000 times.

The application belongs to the field of bioengineering, and particularly relates to a Clostridium edematis subunit protein and a preparation method and application thereof. The Clostridium edematis subunit protein provided by the application is a protein formed by artificial design and combination of recombination expression optimization according to the sequences of main immunogenic proteins of Clostridium edematis, such as main core antigenic fragment sequences of cytotoxin CctA, flagellin FliA and cell wall surface anchoring protein, etc. The protein is prepared into a Clostridium edematis subunit vaccine as an antigen, can effectively resist the infection of Clostridium edematis, can maximize the safety and effectiveness of the vaccine, and can simplify the preparation process and reduce the production cost.

The application provides a kind of chimeric flagellin and its application.The chimeric flagellin is formed by the chimeric of A type Seneca virus VP2 fragment and Escherichia coli Nissle 1917 flagellin FliC through Linker (GGGGS) 3, which realizes the soluble expression of chimeric flagellin in Escherichia coli system, and the yield can reach 200 mg / L at most. And through the detection of in vitro Caco2 and Raw264.7 cell models, the recombinant chimeric flagellin shows good TLR5 receptor activity, which can effectively activate TLR5 signal pathway.

The application belongs to the technical field of plant disease prevention and treatment, and discloses a flagellin and application of the flagellin in a plant immune elicitor. The flagellin is separated and extracted from Bacillus amyloliquefaciens Ba168, has good bacteriostatic activity, and has an immune induction effect on tobacco. The application realizes mass expression and purification of the flagellin by using genetic engineering technology. The flagellin can induce a plant immune response, can induce plant cell necrosis, plant active oxygen burst, and an increase of plant defense enzyme activity. The flagellin can induce plant resistance to PVX, and provides a new type of elicitor material for prevention and treatment of PVX.

This application discloses a carrier protein with adjuvant function and its application. The carrier protein with adjuvant function in this application has the sequence shown in SEQ ID NO.1. The carrier protein with adjuvant function in this application is a partial sequence extracted from the flagellin FliC of Burkholderia cepacia, which retains the adjuvant activity of flagellin while reducing its immunogenicity and antigenicity; when applied to fusion proteins or small molecule drugs, it can enhance their immunogenicity, which is of great significance for subunit vaccines, peptide vaccines, or small molecule drug carriers.

The invention discloses lactococcus lactis for promoting intestinal colonization. The lactococcus lactis is recombinant lactococcus lactis capable of expressing flagellin. Accordingly, a corresponding construction method is also established, that is, the flagellin gene is subcloned to a lactococcus lactis expression vector, and then recombinant plasmids are transformed into lactococcus lactis to obtain the lactococcus lactis. Research shows that the recombinant lactococcus lactis with the mFliC3 and the rFliC3 shows an adhesion promoting function in a cell co-culture experiment, and in a sterile mouse experiment, although the bacteria discharge time of the mFliC3 and the rFliC3 is longer than that of an empty vector group, the bacteria discharge time of the rFliC3 is longer than that of the mFliC3. Therefore, both the mFliC3 and the rFliC3 have the function of promoting field planting, and the field planting time of the rFliC3 is longer. Therefore, the intestinal colonization promoting lactococcus lactis disclosed by the invention can be used as probiotics or functional bacteria, is particularly applied to related food or health care products, and can be used for reducing the supplementing times or prolonging the supplementing interval time.

The invention discloses a novel TCR-CAR plasmid for expressing flagellin, a CAR-T cell, a preparation method of the novel TCR-CAR plasmid and the CAR-T cell, and application of the novel TCR-CAR plasmid and the CAR-T cell in tumor resistance, and belongs to the technical field of CAR-T cells. By means of the innovative structural design, after CAR recognizes a target antigen, expression and secretion of flagellin can be started along with activation of T cells, the flagellin is released to a tumor site, after an activation signal disappears, release of the flagellin gradually fades away, and after the target antigen is recognized again, a new round of secretion and release is started. The technical problems that in the prior art, release of flagellin polypeptide cannot be accurately controlled, engineering immune cells release randomly, and then the anti-tumor effect cannot be achieved due to the fact that the engineering immune cells cannot effectively enter the tumor in the application of treating solid tumors can be effectively solved.

This invention relates to the field of biological protein application technology, and discloses the application of flagellin hypervariable region H as an intramolecular adjuvant in vaccine preparation. The amino acid sequence of flagellin hypervariable region H is shown in SEQ ID NO.3, and the nucleotide sequence of the gene encoding flagellin hypervariable region H is shown in SEQ ID NO.4. This invention also discloses an expression cassette, recombinant vector, recombinant cells, or recombinant bacteria containing the gene encoding flagellin hypervariable region H. This invention uses the aforementioned flagellin hypervariable region H as an intramolecular adjuvant in vaccine preparation. This invention connects flagellin hypervariable region H to an antigenic epitope to obtain a fusion protein. Compared with existing technologies, the solubility and activity of the fusion protein are improved. Simultaneously, this invention overcomes the expression form of inclusion bodies in existing E. coli expression systems, eliminating the need for subsequent refolding and assembly, thus simplifying the vaccine preparation process.

The application discloses a method for improving the anti-styrene stress ability of Gluconobacter oxydans, and belongs to the technical field of genetic engineering and microbial engineering. Firstly, the adaptability evolution of wild-type Gluconobacter oxydans to styrene is carried out by gradient addition of styrene in a culture medium, and finally, the evolved strain ST significantly improves the tolerance of Gluconobacter oxydans to styrene. Secondly, the knocking out of flagellin FlgE in Gluconobacter oxydans also improves the stress ability of Gluconobacter oxydans to styrene; the growth performance of the gene deletion Gluconobacter oxydans ST-ΔflgE prepared by the method of the application after 10h culture under 15g / L styrene is far better than that of the wild-type Gluconobacter oxydans.