67 results about "Pilin" patented technology

The invention provides chimeric proteins comprising a non-toxic Pseudomonas exotoxin A sequence and a Type IV pilin loop sequence, wherein the Type IV loop sequence is inserted within the non-toxic Pseudomonas exotoxin A. The invention also provides polynucleotides encoding the chimeric proteins, and compositions comprising the polynucleotides or the chimeric proteins. The invention also provides methods for using the chimeric proteins, polynucleotides and compositions of the invention.

Methods and compositions for inducing an immune response against Pseudomonas aeruginosa are provided herein. In one aspect, the invention provides a chimeric immunogen, comprising a receptor binding domain, a translocation domain, and a Pseudomonas pilin peptide comprising an amino acid sequence that is TAADGLWKCTSDQDEQFIPKGCSK (SEQ ID NO.:1), wherein the chimeric immunogen, when administered to a subject, induces an immune response in said subject that is effective to reduce adherence of a microorganism that expresses said Pseudomonas pilin peptide to epithelial cells of said subject. In other aspects, the invention provides nucleic acids encoding chimeric immunogens of the invention, kits comprising chimeric immunogens of the invention, cells expressing chimeric immunogens of the invention, and methods of using chimeric immunogens of the invention.

Methods and compositions for inducing an immune response against Pseudomonas aeruginosa are provided herein. In one aspect, the invention provides a chimeric immunogen, comprising a receptor binding domain, a translocation domain, and a Pseudomonas pilin peptide comprising an amino acid sequence that is TAADGLWKCTSDQDEQFIPKGCSK (SEQ ID NO.:1), wherein the chimeric immunogen, when administered to a subject, induces an immune response in said subject that is effective to reduce adherence of a microorganism that expresses said Pseudomonas pilin peptide to epithelial cells of said subject. In other aspects, the invention provides nucleic acids encoding chimeric immunogens of the invention, kits comprising chimeric immunogens of the invention, cells expressing chimeric immunogens of the invention, and methods of using chimeric immunogens of the invention.

The invention relates to a PCR (polymerase chain reaction) method for specific detection of watermelon acidovorax citrulli. The method designs a pair of primers according to an assumed IV type pilin gene (GenBank: CP000512.1) sequence on the complete genome sequence of a watermelon acidovorax citrulli strain AAC00-1, and a BLAST result indicates that the pair of primers have very good specificity. During PCR detection, the pair of primers not only can eliminate other bacteria, and can distinguish the acidovorax citrulli in watermelons and the acidovorax citrulli in muskmelons. And the pair ofprimers can distinguish an acidovorax citrulli strain in a watermelon from other bacteria accurately and have high specificity.

The invention provides FIG. 1 novel antimicrobial chemical entities based on a nitrothiazolide backbone that exhibit antibacterial and antiparasitic action against a wide range of human pathogens. The new classes of compounds show extended action against Gram positive bacteria including MRSA drug resistant pathogens. In the Gram-positive organisms, they specifically target and functionally inhibit microbial attachment to surfaces and biofilm formation. In Gram-negative bacteria, including enteroaggregative E. coli strains, these compounds function as pilicides by inhibiting the assembly of pilin subunits into adhesive filaments. Several of these compounds show potent antimicrobial action against Gram positive bacteria, perhaps involving novel targets. Many of the benzothiophene derivatives exhibit antimicrobial activity in the low micrograms per ml range and in blocking biofilm formation in the nanomolar range; ranges considered are well within the range of utility as therapeutics.

A protein construct comprising a pilus protein portion, preferably a structurally stabilized pilus-protein, and an additional, or effector, portion other than a pilus protein or chaperone and wherein said effector portion serves to stabilize the pilus protein portion and to confer a therapeutic activity, such as vaccine activity or anti-microbial or anticancer activity, on the protein construct is disclosed. Such effector portion commonly comprises a donor strand complementary segment capable of structurally stabilizing a pilus protein subunit and attaching the auxiliary portion to said subunit to form the pilus protein analog of the invention. Methods of using said protein constructs are also disclosed as well as the formation and use of analogs comprising fragments of a pilus protein linked to effector components to produce immunogenic and / or therapeutic activity.

The present invention relates to a composition and method for preventing or inhibiting biofilm formation on biotic or abiotic surfaces. The composition comprises a peptide based on the C-terminal receptor binding domain of Pseudomonas aeruginosa type IV pilin, which binds to an abiotic surface (e.g., steel, plastic) with high affinity and prevents binding of a variety of P. aeruginosa strains to the surface. The inventive composition represents a non-toxic inhibitor for biofilm formation, particularly on an abiotic surface, which is responsible for a large number of problematic diseases and massive economic losses. The inventive method is useful as a safe and environmentally friendly means of modifying a surface of a variety of biomedical, nanotechnological, and biotechnological devices or articles.

The invention provides chimeric proteins comprising a non-toxic Pseudomonas exotoxin A sequence and a Type IV pilin loop sequence, wherein the Type IV loop sequence is inserted within the non-toxic Pseudomonas exotoxin A. The invention also provides polynucleotides encoding the chimeric proteins, and compositions comprising the polynucleotides or the chimeric proteins. The invention also provides methods for using the chimeric proteins, polynucleotides and compositions of the invention.

The present invention relates to engineered Clostridium difficile type IV pilin (tfp) genes, type IV pilin proteins which can serve as a diagnostic marker for identification of patients infected with C. difficile, and vaccines comprising type IV pilin proteins, antigenic fragments and variants thereof for therapeutic interventions.

The invention discloses ETEC (enterotoxigenic escherichla coli) yolk antibody powder and a preparation method thereof. The method comprises following steps: escherichia coli strains are cultured in a solid culture medium at 35-40 DEG C for 18-24 h; after culture, pilin is extracted coarsely from the solid culture medium and is purified; the purified pilin and equal volume of a freund's adjuvant are mixed, an oil-emulsion vaccine is formed and used for immunizing a primiparous hen, and serum and a yolk antibody are collected after immunization; the yolk antibody is subjected to spray drying through a powder sprayer, and the yolk antibody powder is prepared. Analysis of the titer of the immunized yolk antibody with ELISA (enzyme-linked immunosorbent assay) discovers that higher-titer yolk antibody can be obtained by using purified pilin as an antigen for immunization. After the yolk antibody powder is prepared from the yolk antibody through spray drying by the powder sprayer, the antibody titer is only reduced by one degree of multiple proportions, the yolk antibody has very good high-temperature stability, and a power preparation process adopting a spray drying method is applicable to production of the yolk antibody powder.

The present disclosure provides coding and amino acid sequences for a Campylobacter jejuni pilus protein (and from other species as well). This protein, when administered to a human or animal, elicits the expression of an immune response to Campylobacter jejuni, with the result that colonization and / or infection by this organism is reduced. Recombinant protein or biofilm material comprising the pilus protein is formulated into immunogenic compositions, especially for mucosal administration. Thus, the present invention provides methods for improvement of the microbial quality of food products including poultry, eggs, meat and dairy products, and indirectly of plant foods that may come in contact with agricultural waste, either from fertilization or from irrigation water.

The invention relates to the technical field of microbe, and relates to a lactobacillus rhamnosus immunomagnetic beads electrochemical sensor detection method. The method is characterized in that immunomagnetic beads having a lactobacillus rhamnosus specific pilin subunit SpaA antibody and an electrochemical sensor are combined for performing quantitative determination on lactobacillus rhamnosus in a probiotic product. The method has the advantages of simple operation and no requirement of culture, a detection period is 2-3 hours, the method is convenient for popularization and usage, and is suitable for rapid quantitative determination and quality evaluation on lactobacillus rhamnosus in the probiotic product and a fermented product.

The present invention relates to compositions comprising Haemophilus influenzae Protein E and Pilin A. More particularly, the present application relates to fusion proteins and immunogenic compositions comprising Protein E and PilA, vaccines comprising such immunogenic compositions and therapeutic uses of the same.

The invention relates to the technical field of biologic pharmacy, and discloses a method for removing pertussis component pilin 2 / 3 endotoxin. The method comprises the steps that after bordetella pertussis is fermented and cultured, thalli are centrifugally collected, thalli are extracted through a urea phosphate buffer solution, a supernate is centrifugally collected, and PEG-8000 and ammonium sulfate are used for performing precipitation; precipitates are extracted by the phosphate buffer solution, then, a supernate is centrifugally collected, a Q-Sepharose chromatographic column is adoptedfor processing, and a phosphate buffer solution containing sodium chloride is adopted for eluting the Fim 2 / 3 component. By combining PEG8000 precipitating, ammonia sulfate precipitating and Q-Sepharose chromatographic column processing are combined, endotoxin in pilin 2 / 3 is removed through repeated means, on the premise of not introducing allogenic materials, the effective ingredient Fim 2 / 3 biologic activity and recycling rate are kept, and meanwhile the endotoxin in Fim 2 / 3 can be removed to 10 EU / mg or lower.

The invention provides novel antimicrobial chemical entities based on a nitrothiazolide backbone that exhibit antibacterial and antiparasitic action against a wide range of human pathogens. The new classes of compounds show extended action against Gram positive bacteria including MRSA drug resistant pathogens. In the Gram-positive organisms, they specifically target and functionally inhibit microbial attachment to surfaces and biofilm formation. In Gram-negative bacteria, including enteroaggregative E. coli strains, these compounds function as pilicides by inhibiting the assembly of pilin subunits into adhesive filaments. Several of these compounds show potent antimicrobial action against Gram positive bacteria, perhaps involving novel targets. Many of the benzothiophene derivatives exhibit antimicrobial activity in the low micrograms per ml range and in blocking biofilm formation in the nanomolar range; ranges considered are well within the range of utility as therapeutics.

The invention provides an extraction method for an escherichia coli pilus antigen used for preparing a yolk antibody. The extraction method comprises the steps of (1) culturing escherichia coli, collecting thalli, breaking and dissolving the thalli and preparing a primary extract of escherichia coli thalli; 2) purifying the primary extract by a 4FF gel chromatographic column to obtain a purified solution containing escherichia coli pilus protein; and 3) removing endotoxin in the purifiedsolution of escherichia coli pilus protein by using acetone precipitation to obtain the escherichia coli pilus antigen. Pilus protein with relatively high purity is obtained by utilizing gel filtration chromatography; and factors such as endotoxin released by the thalli which are possible to interfere an immune effect are removed. The prepared subunit vaccine immunized laying hens gain high-titer antibodies; and new ideals are provided for preventing and treating related diseases.

Compositions for eliciting an immune response against Gram-negative bacterial infections and methods of making such compositions are provided. The composition comprises glycosylated pilin, the pilin being glycosylated with the O-antigen of a target Gram-negative bacteria of interest. Methods of eliciting an immune response by administration of such compositions are also provided.

The invention discloses an avian Escherichia coli type 1 pilin expression recombinant bacteria, a construction method and application thereof. The strain is imported with avian Escherichia coli type 1 fimbriae operon gene recombinant plasmids and is constructed with the following steps: 1. PCR augmentation of avian Escherichia coli type 1 fimbriae structure gene and each functional gene; and 2. recombinant plasmid construction. The invention further discloses an avian Escherichia coli type 1 fimbriae engineered vaccine. The avian Escherichia coli type 1 pilin expression recombinant bacteria has expression product with protuberant fimbriae, good immunogenicity, high culture expression yield, low cost and easy implementation of industrial vaccine production.

The invention provides PA (pseudomonas aeruginosa) recombinant protein POP, as well as a preparation method and an application thereof. The recombinant protein is formed by sequential connection of a PA III type secretory system component protein PcrV fragment, a PA outer membrane lipoprotein OprI fragment and a PA IV type pilin PilA fragment through a linker. The recombinant protein has a general formula: PcrV fragment-(Linker A)m-Oprl fragment-(Linker B)n-PilA fragment, wherein sequences of the Linker A and the Linker B are respectively independently selected from one of GGGGS, GGSGG and YAPVDV; values allowed for m and n are 1, 2, 3 and 4 independently and respectively. The invention also provides the preparation method and the application of the recombinant protein. Subunit vaccine prepared from the recombinant protein can stimulate the organism to generate high-titer IgG antibodies; the recombinant protein can be used for preparing diagnostic drugs, preventive drugs or therapeutic drugs.

The invention belongs to the technical field of veterinary biology, and in particular relates to a yolk antibody for preventing and treating colibacillosis and a preparation method and application of the yolk antibody. The escherichia coli yolk antibody is prepared by using a mixed solution of extracted avian pathogenic escherichia coli O78 outer membrane protein and pilin as antigen immune chicken; and experiments show that the yolk antibody is safe and efficient and has a good protective effect on homogenous and multiple alien escherichia coli. The yolk antibody can be used for preventing and treating multi-serotype colibacillosis, and can also be used as an additive of health-care products, food and feed or used for preparing preparations for detecting related diseases caused by escherichia coli infection.

The present invention provides synthetic chimeric fimbrin peptides which induce an immunogenic response in animals to non-typable Haemophilus influenzae and that do not require tedious purification techniques. The synthetic chimeric fimbrin peptides reduce the severity of otitis media caused by Haemophilus influenzae. The synthetic chimeric fimbrin peptides are synthesized using commercially available peptide synthesizers. The synthetic chimeric fimbrin peptides comprises three peptide units. The first peptide unit is a subunit of the fimbrin protein. Preferably, the fimbrin subunit is comprised of the amino acids of Sequence ID No. 1 or Sequence ID No. 2. The second peptide unit is a t cell epitope, and preferably has the amino acid sequence of SEQ ID NO. 3. The third peptide unit is a linker peptide unit which joins the first and second peptide unit. The linking sequence preferably has from about 2 to about 15 amino acids, more preferably from about 2 to about 10 amino acids, most preferably from about 5 to about 6 amino acids. The synthetic chimeric fimbrin peptides are useful immunognes against NTHi and also useful as laboratory tool for detecting antibodies in sera. The invention also relates to an immunogenic composition containing the synthetic chimeric fimbrin peptides and a pharmacologically acceptable carrier.

The present invention relates to engineered Clostridium difficile type IV pilin (tfp) genes, type IV pilin proteins which can serve as a diagnostic marker for identification of patients infected with C. difficile, and vaccines comprising type IV pilin proteins, antigenic fragments and variants thereof for therapeutic interventions.

The invention provides a preparation method for a piglet enterotoxigenic escherichia coil-resistant egg yolk antibody, and belongs to the technical field of antibody preparation. The preparation method for the piglet enterotoxigenic escherichia coil-resistant egg yolk antibody adopts an existing piglet enterotoxigenic escherichia coil gene engineering inactivated vaccine and adopts piglet escherichia coli diarrhea diseased pig intestinal tract isolate enterotoxin and adhesive pilin to perform muscle immune on an SPF laying hen, in an immunologic process, Enterococcus faecium Anp01 CCTCC M 200919 is fed and a traditional Chinese medicine composition is smashed for feeding serving as an immunopotentiator, eggs of 2-6 weeks of immune infectious enteritis are collectd, an egg yolk aqueous extract solution is extracted, the egg yolk aqueous extract solution is purified by using a freeze-thaw method, so as to obtain the egg yolk antibody IgY. The antibody has a practical value to emergency treatment of diarrhea caused by piglet enterotoxigenic escherichia coil, and has important significance on reducing economic loss of pig industry and promoting development of a new veterinary biological drug.

The invention provides novel antimicrobial chemical entities based on a nitrothiazolide backbone that exhibit antibacterial and antiparasitic action against a wide range of human pathogens. The new classes of compounds show extended action against Gram positive bacteria including MRSA drug resistant pathogens. In the Gram-positive organisms, they specifically target and functionally inhibit microbial attachment to surfaces and biofilm formation. In Gram-negative bacteria, including enteroaggregative E. coli strains, these compounds function as pilicides by inhibiting the assembly of pilin subunits into adhesive filaments. Several of these compounds show potent antimicrobial action against Gram positive bacteria, perhaps involving novel targets. Many of the benzothiophene derivatives exhibit antimicrobial activity in the low micrograms per ml range and in blocking biofilm formation in the nanomolar range; ranges considered are well within the range of utility as therapeutics.

The present invention relates to the use of mutant glycoproteins from pathogenic bacteria lacking one or more phosphorylcholine and / or glycosylation post-translational modifications as immunogens. These post-translational modifications act as masking structures that elicit an immune response which does not confer protection on an infected individual. The removal or modification of these masking structures alters the protein such that it elicits a stronger immune response to the protein and / or the bacterial pathogen. Particular examples are pilin proteins and nitrite reductase glycoproteins of Neisseria bacteria.

The invention discloses a construction method of an anti-oxidation adhesion type genetically engineered bacterium, the engineering bacterium and application of the engineering bacterium, and relates to the technical field of construction of the anti-oxidation adhesion type genetically engineered bacterium. According to the invention, superoxide dismutase and catalase are subjected to constitutive overexpression in a probiotic chassis by using a synthetic biological technology, and meanwhile, intestinal trefoil factor peptide fused pilin is subjected to overexpression. According to the present invention, the research results show that the active oxygen scavenging ability of the anti-oxidation adhesion type gene engineering probiotic is significantly improved, the adhesion ability of the anti-oxidation adhesion type gene engineering probiotic in the animal intestinal tract is significantly enhanced, and the radiation enteritis symptom caused by the whole abdomen ionizing radiation irradiation can be effectively relieved; the strain can be used as a potential safe viable bacterium medicine for preventing and treating radiation enteritis and other active oxygen related enteritis. The anti-oxidation adhesion type genetic engineering probiotics can be prepared into viable bacteria preparations such as viable bacteria suspension, dairy products, viable bacteria powder, bacterial tablets or capsules, can be proliferated and colonized in human intestinal tracts after being orally taken, and is applied to prevention and treatment of radiation enteritis.

A Tularemia vaccine proposes to induce antibodies which through their binding to the target proteins of the vaccine strategy will inactivate the proteins essential to mediating the secretory function of the pilus proteins of the “type IV pilus proteins” or Tfp proteins, thereby impairing the formation of the pilus proteins on the surface of the FT which are needed for binding and entry of the FT into macrophages, and will inhibit the secretion of several factors of FT which are necessary for the virulence of the FT, and which are missing in attenuated strains of FT used in current attenuate strain vaccination strategies. The vaccine may be a mixture of DNA plasmids encoding TAA / ecdCD4OL protein vaccines in which the TAA is comprised of fragments of the following Tularemia proteins: Tfp, pilA, pilC, pilT, and pilQ,

The present invention relates to engineered Clostridium difficile type IV pilin (tfp) genes, type IV pilin proteins which can serve as a diagnostic marker for identification of patients infected with C. difficile, and vaccines comprising type IV pilin proteins, antigenic fragments and variants thereof for therapeutic interventions.

The present invention relates to compositions comprising Haemophilus influenzae Protein E and Pilin A. More particularly, the present application relates to fusion proteins and immunogenic compositions comprising Protein E and PilA, vaccines comprising such immunogenic compositions and therapeutic uses of the same.

The invention belongs to the technical field of biopharmaceutics, and in particular to a group B Neisseria meningitidis recombinant pilin Fim, and a preparation method and application thereof. The present market lacks a vaccine which directly blocks adhesion of Neisseria meningitidis pilin to prevent meningitis. The invention provides a group B Neisseria meningitidis recombinant pilin Fim gene with a nucleotide sequence shown as SEQID NO:1. The group B Neisseria meningitidis recombinant pilin Fim gene is first found to be a key molecule for adhesion and colonization of Neisseria meningitidis.The group B Neisseria meningitidis recombinant pilin Fim gene is cloned into a protein expression vector and then is expressed on a large scale, and the recombinant protein is prepared after purification, wherein the purity of the recombinant protein is high and reaches 90% or above. The repeatability is good and the recovery rate is high. The protein can be used as a vaccine candidate to preventinfection of group B Neisseria meningitides, has the advantages of soluble expression, easy purification, high purity and simple preparation method, and has notable economic benefits.