63 results about "Bacterial capsule" patented technology

Precipitated bacterial capsular polysaccharides can be efficiently re-solubilised using alcohols as solvents. The invention provides a process for purifying a bacterial capsular polysaccharide, comprising the steps of (a) precipitation of said polysaccharide, followed by (b) solubilisation of the precipitated polysaccharide using ethanol. CTAB can be used for step (a). The material obtained, preferably following hydrolysis and sizing, can be conjugated to a carrier protein and formulated as a vaccine. Also, in vaccines comprising saccharides from the serogroups A and C, the invention provides that the ratio (w/w) of MenA saccharide:MenC saccharide is >1.

The present invention relates to a polyvalent bacterial capsule polysaccharide-protein conjugate combined vaccine preparation, in particular, it is a combined vaccine containing group A, group C, group Y and group W135 epidemic cerebrospinal meningitis coccal capsule polysaccharide-protein conjugate and b type haemophilus influenzal capsule polysaccharide-protein conjugate.

The present invention relates to a streptococcus pneumoriae polysaccharide-outer membrane protein combined vaccine and its preparation method, belonging to the field of streptococcus pneumoriae vaccine. The main antigen component of said vaccine is a streptococcus pneumoriae capsular polysaccharide-outer membrane protein combined product obtained by covalently connecting the capsular polysaccharide produced by streptococcus pneumoriae with its outer membrane protein. Said capsular polysaccharide is the capsular polysaccharide of one or several kinds of streptococcus pneumoriae, its molecular weight is about 200-500 KDa, every polysaccharide molecule has about 300-700 repeating units. The outer membrane protein is the outer membrane protein of one or several kinds of streptococcus pneumoriae, its molecular weight is about 30-100 Kda. Said vaccine can be used for preventing or curing the diseases induced by streptococcus pneumoriae.

Methods are described for improvement of the serum half life of therapeutic nucleic acids by 3′ conjugation to useful target proteins, or other large molecules with useful function. In one embodiment, a 3′ A, C or G overhang is added to ds-DNA and the primary amines conjugated using biocompatible bifunctional linkers to proteins. The resulting nucleic acid-3′-conjugates are serum nuclease-resistant and retained in vivo for long periods without rapid kidney clearance. Further, the choice of conjugate imparts additional functionality to the nucleic acid-3-conjugate. For example, if the protein in the DNA-protein conjugate is the first component of the complement cascade (Clq or Clqrs) and the DNA aptamer has been developed against surface components of a target cell, it can be used to treat bacterial or parasitic infections and cancers. If the protein is serum albumin or another common (nonimmunogenic) blood protein and the aptamer is directed against a toxin or venom, the aptamer-protein conjugate can be used as an antidote that binds and neutralizes the toxin or venom. Similar DNA (aptamer)-nanotube, -enzyme, and -toxin conjugates could also be used to target and selectively kill bacteria, parasites, and cancer cells in vivo. If the protein is an Fc antibody fragment or C3b protein from the complement system and the aptamer is developed against a bacterial cell capsular material, other cell surface component or viral cell surface component, then the aptamer-3′-protein conjugate can aid in opsonization of the target cells or viruses by phagocytic leukocytes.

This invention relates generally to the discovery of a method of inhibiting, preventing or treating bacterial infections. The invention also relates to a method of inhibiting bacterial capsule biogenesis.

The invention relates to a bacterial capsule staining method, comprising the steps: 1) dropping a droplet of sterile water on a glass slide; 2) bacteria culturing for 24 to 48 hours, selecting the bacteria with a transfering loop, and slowly coating the glass slide in water moisture state spirally from the center to the outside to keep the capsule complete; 3) natural drying or blow drying with electric wind; 4) dropping 2-3 droplets of pure methanol, fixing for 1-2 minutes, and inclining to remove the methanol; 5) dropping carbolic acid fuchsin to stain for 1-3 minutes, and covering a cover glass; and 6) absorbing the liquid surrounding the cover glass with a piece of absorbent paper, inclining the slide glass to absorb partial staining liquid under the cover glass. By improvement on the slide making method, the staining method and the operation steps, the completeness of bacterial capsule is effectively kept, the growing state of all the bacterial capsule in the specimen can be clearly displayed, the operation difficulty of capsule staining is obviously reduced, and the reliability of bacterial capsule recognition is improved.

A method for measuring content of free protein in solder of bacterial capsule protein-polysaccharide includes determining out carrier protein concentration in bacterial capsule protein-polysaccharide solder to be detected by utilizing Lowry manner, using carrier protein corresponding to solder to be detected as sample one and using solder to be detected as sample two, using high efficiency gel chromatography to obtain chromatogram of sample one and two separately then applying external-labeling means to calculate out content of free protein.

The invention relates to the fungal polysaccharide preparation field, and concretely relates to a preparation method of Cryptococcus Neoformans capsular polysaccharide. The invention provides a method for preparing the Cryptococcus Neoformans capsular polysaccharide through utilizing cetyl trimethyl ammonium bromide. The method comprises the following steps: 1, processing a secretion supernatant containing the capsular polysaccharide through using calcium acetate and glacial acetic acid, and precipitating through using ethanol to prepare a crude capsular polysaccharide; and 2, processing a solution of the crude capsular polysaccharide through using the cetyl trimethyl ammonium bromide to obtain the purified capsular polysaccharide. The method has the advantages of simplicity, high efficiency, low cost, and effective avoiding of the use of toxic and harmful chemical reagents.

Compositions and methods for the detection, prevention, or treatment of anthrax or other infectious diseases. In one aspect, the present invention provides methods of immunizing humans or animals against Bacillus anthracis or other capsulated pathogens. The methods include administering a capsular polypeptide of a pathogen of interest and a CD40 agonist to a human or animal. The capsular polypeptide or the CD40 agonist is administered in such an amount or frequency that an immunoprotective response can be elicited in the human or animal against the pathogen of interest. In another aspect, the present invention provides methods of using passive immunization with anti-capsular polypeptide antibodies to prevent or treat infections caused by Bacillus anthracis or other pathogens. In yet another aspect, the present invention provides methods useful for diagnosis of anthrax by detection of capsular polypeptide in serum or other biological samples.

The invention discloses a method for rapid purification of bacterial capsular polysaccharide. The method can fast remove pollutants comprising proteins and nucleic acid from a specific bacterial fermentation broth and retains purified capsular polysaccharide. The method comprises the following steps of fermentation of capsular polysaccharide-containing bacteria, acid adjustment of a pH value of a fermentation broth, precipitation of thalli and impurities of the fermentation broth, centrifugation, microfiltration, ultrafiltration condensation and filter wash so that a crude bacterial capsular polysaccharide solution is obtained. A basic principle of the method comprises that through acid adjustment, a pH value of a fermentation broth is in a range of 3 to 5 so that thalli having whole shapes and other impurities are removed and capsular polysaccharide is retained in the fermentation broth and thus purification of capsular polysaccharide is realized. The method can be used for purification of capsular polysaccharide of pneumococcocci, haemophilus influenzae type b, epidemic meningococcocci and typhoid bacillus.

The invention relates to a sedum rhizosphere cadmium-resistant bacillus sp. BCd5 strain and its screening method. The invention specifically relates to a sedum rhizosphere cadmium-resistant bacillus sp. BCd5 strain which has been preserved in the China General Microbiological Culture Collection Center (CGMCC) on December 1st, 2014 with the preservation number being CGMCC NO. 10077. 16S rDMA gene sequence of the strain is as shown in the Figure 1. The strain is a Gram-negative bacterium, aerobasilus having flagellum and capsule. The strain has high resistance to cadmium and has practical significance and important engineering application value in plant-microbial remediation of cadmium-contaminated soil.

The invention provides a preparation method of artificial capsule type multifunctional particles, and belongs to the field of surface water pollution treatment. According to the preparation method, ore is used as a carrier, microorganisms are loaded on the ore by adding a binder and a flocculant, and meanwhile, mineral particles are coated with a binder layer, a flocculant layer and a high-efficiency degrading bacterium layer to form structures similar to bacterial capsules. The capsule structure not only can protect microorganisms in mineral particles, but also has the capability of immobilizing external efficient degrading bacteria. Minerals, a binding-flocculating material and microorganisms in the multifunctional particles coordinate with one another, comprehensive utilization is realized, the advantages of each components are maintained, optimal adjustment is performed according to different pollution characteristics of water body, the floating water purification capacity is effectively improved, the effects of energy conservation, emission reduction and resource utilization are achieved, and the method can be widely applied to pollution treatment of rivers, lakes and reservoirs and has good market application prospects.

The invention belongs to the field of biological products and relates to a preparation method of pneumococcal capsular polysaccharide. According to the preparation method, pneumococcus inactivated andclarified fermentation liquid as an initial feed liquid is used for purification and preparation by steps as follows: ultrafiltering and concentrating the clarified fermentation liquid, adding CTAB to form a precipitate with polysaccharide, performing centrifugation to collect the precipitate, and performing depolymerization by sodium chloride and chromatography purification to finally obtain pneumococcal capsular polysaccharide; viscosity control comprises steps of ultrafiltering, clarifying and filtering, precipitation by CTAB, depolymerization by sodium chloride and chromatography.

The invention discloses a screening method of feed probiotics capable of preventing chicken's infection caused by clostridium perfringens. The method comprises following steps: screening chickens infected by clostridium perfringens in clinic and disease resistant chickens; collecting the intestinal chyme samples; carrying out high throughput sequencing on microbial floras in the intestinal tractsof infected chickens and disease resistant chickens; comparing and analyzing the microbes in the intestinal tracts of infected chickens and disease resistant chickens through a metagenomics method; and screening and identifying probiotics capable of preventing infection caused by clostridium perfringens. According to the method, the metagenomics big data of floras in the intestinal tracts of infected chickens and disease resistant chickens is analyzed; probiotics capable of preventing infection caused by clostridium perfringens are selected from massive microbial floras in the intestinal tracts of disease resistant chickens, the screened probiotics are from same animals, the rejection between probiotics and a host is avoided, moreover, the screening is targeted, aimless screening is avoided, and a large amount of labor, resources, and time is saved.

The invention discloses a method for purifying bacterial capsular polysaccharide. The method comprises the following steps: treating a streptococcus pneumonia fermentation culture solution with an inactivator, adding acid for regulating the pH value to be less than or equal to 6.5, precipitating impurities, centrifuging, and carrying out microfiltration for removing precipitates, so that polysaccharide clarified liquor is obtained; carrying out ultrafiltration liquid change on the polysaccharide clarified liquor by adopting a membrane with the molecular weight cut-off of 10-100KDa and concentrating, and carrying out chromatography on the obtained capsular polysaccharide solution by combining compound type ion exchange chromatography and hydroxyl phosphate grey salt chromatography, so that the purified capsular polysaccharide solution is obtained; and concentrating and carrying out ultrafiltration on the capsular polysaccharide solution, and carrying out sterile filtration and storing. By combining the compound type ion exchange chromatography and the hydroxyl phosphate grey salt chromatography, the contents of protein and nucleic acid impurities can be effectively reduced to be lower than 1%, the quality of a purified polysaccharide product is higher than European Union pharmacopoeia standard requirements, and the recovery rate is more than 60%. The method disclosed by the invention has the advantages that the technology is simple and rapid, the enlargement is easy, and industrial production can be realized.

The invention discloses a high-throughput screening method of a colloid bacillus salt tolerant mutant strain, which comprises the following steps: firstly, after being activated in nitrogenfree medium, colloid bacillus is placed in a liquid culture medium for restraining the growth of capsula, and cultured to the logarithmic growth phase, thallus is centrifuged and collected, physiological saline or phosphate buffer is used for suspending the thallus, and the concentration of the thallus is adjusted to 10<4>-10<5>cfu ml<-1>; secondly, Co<60> is used for inducing the thallus suspending liquid, and the amount of a mutagenic agent is 1-8kGy; the thallus suspending liquid after being induced is centrifuged to obtain the induced thallus; thirdly, after being diluted to 10<3>-10<4>cfu ml <-1> by using the liquid screening culture medium, the mutate thallus is cultured after being separately packaged into a culture plate with 96 holes for the high-throughput screening, after the secondary screening is performed, the salt tolerant mutant strain is obtained. In the invention, nuclear radiation inducement and the high-throughput screening method are used for obtaining the salt tolerant mutant strain of the colloid bacillus, so as to provide the strain for the rehabilitation of greenhouse soil.

Production of protein conjugate vaccines by use of transpeptidase enzymes, such as sortase enzymes. For example, homogenous immunoconjugates (e.g., a population of molecules having the same structure) formed by conjugating an antigenic polypeptide and a bacterial capsule component are provided. In certain aspects, methods for generating an immune response to B. anthracis by use of protective antigen-PDGA immunoconjugates are provided.

Disclosed are a sulfate reducing bacterium with a polyacylamide degradation function and applications of the bacterium. The invention relates to a microorganism and applications of the microorganism. The invention firstly put forward the sulfate reducing bacterium with the polyacylamide degradation function, the bacterium is named as Salmonella Daqing MF, namely, Salmonella Daqing MF (preservation code: CGMCC No.1910), preserved in the China General Microbiological Culture Collection Center, and belonging to the Salmonella Genera and the Salmonella Daqing Species in taxonomy. The bacterium belonging to the Gram negative short-bacilli is covered by a capsule and is obligate anaerobic, variable in size bound in 0.35Mu-0.7Mum width multiplied by 0.7Mum-1.6Mum length, without flagella on the bacterium body; when growing on a solid medium, the bacterium develops into a black round colony, smooth in surface and irregular in edge.

Several species of bacteria capable of invasive infections, such as S. pyogenes, S. equi and P. multocida, contain hyaluronic acid (HA) in their capsules. Bacterial species such as Staphylococcus aureus and related Staphylococci have capsules that contain acidic polysaccharides. Bacterial capsule or bacterial surface binding peptides were synthesized and tested in a culture model of invasive bacterial infections, specifically translocation through polarized keratinocyte cultures. The peptides reduced the translo-cation of a variety of bacterial species, with a concomitant increase in bacterial internalization by the keratinocytes. In vivo, subcutaneous inoculation of encapsulated GAS treated with peptides delayed bacterial dissemination. In a mouse surgical wound control model infected with S. aureus, treatment with peptides reduced the numbers of bacteria and inflammation at the wound site.

The invention relates to a culture method for increasing the yield of the capsule of avian pasteurella multocida. The culture method comprises the following steps: (1) inoculation: inoculating avian pasteurella multocida into a triangular flask with a culture medium; (2) shake culture: putting the triangular flask after inoculation in the step (1) into an incubator to undergo shake culture at a temperature of 36-38 DEG C for 5.5-6.5 hours, wherein the shaker speed is 180-220rpm; (3) stationary culture: putting the triangular flask after shake culture in the step (2) into the incubator to stand, wherein the culture temperature is 36-38 DEG C and the stationary culture time is 7.5-8.5 hours. The yield of the capsule of avian pasteurella multocida can be obviously increased by adopting the culture method provided by the invention.

The invention discloses a preparation process of crude polysaccharide of an A-group Neisseria meningitidis bacterium capsule. The preparation process of the crude polysaccharide of the A-group Neisseria meningitidis bacterium capsule comprises the following steps that (1) cultivated Neisseria meningitidis is sterilized by utilizing formaldehyde, and supernate is centrifugally collected from a sterilized culture solution; (2) CTAB is added into the supernate till the final concentration of the CTAB is 1.0 g/L, then standing is performed overnight after full stirring, and polysaccharide is centrifugally collected; (3) the precipitated polysaccharide is stirred by using a calcium chloride solution for 3 hours, and the supernate is centrifugally collected; (4) ethyl alcohol is added into the supernate till the final concentration by volume of the ethyl alcohol is up to 20%-25%, standing is performed at the temperature of 2 DEG C to 8 DEG C overnight, the supernate is centrifugally collected, then ethyl alcohol is added into the supernate till the final concentration by volume of the ethyl alcohol is up to 75%-80%, full shaking is performed, standing is performed for more than 18 hours, and a precipitate is centrifugally collected; (5) absolute ethyl alcohol and acetone are adopted to perform washing for three times respectively, and then drying is performed to obtain the crude polysaccharide. The preparation process of the crude polysaccharide of the A-group Neisseria meningitidis bacterium capsule has the advantages of being high in extraction efficiency and the like.