43 results about "Clostridium tetani" patented technology

The invention discloses a method for preparing a tetanus toxoid vaccine. According to the process, with clostridium tetani strains as raw materials, the tetanus toxoid vaccine is prepared through the following steps of: culturing of tetanus toxoid, bacterium liquid separation, ultrafiltration and concentration, salting out, ultrafiltration desalting and the like. According to the method, firstly culture liquid is subjected to virus-free treatment and then refined, so that the porous channel plugging caused by accumulation of thalli and other impurity segments at a plate and frame membrane package during plate and frame filtering to remove thalli is reduced, and the smoothness during filtration is increased; toxoid protein and other allergens in the culture liquid are removed by changing the salting-out method; and as the desalting methods of the culture liquid after concentration and salting out adopt the tangential flow ultrafiltration method, the destruction of antigen caused by shearing of toxoid protein is reduced, and the protein precipitation is avoided. By utilizing the method, the time for preparing the tetanus toxoid vaccine is shortened, and the production efficiency is improved.

The invention relates to a mouse hybridoma cell strain secreting a clostridium tetani bacillus exotoxin monoclonal antibody, wherein the strain has a preservation number of CCTCC (China Center for Type Culture Collection) NO. C201257. The invention further relates to a clostridium tetani bacillus exotoxin monoclonal antibody prepared by the mouse hybridoma cell strain; the invention further provides an Fab antibody, comprising a kappa chain and an Fd chain; the kappa chain and the Fd chain are obtained by amplifying from total RNA (ribonucleic acid) of the hybridoma cell strain. The invention further relates to a medicine for preventing or treating clostridium tetani bacillus infection, comprising the clostridium tetani bacillus exotoxin monoclonal antibody and / or the Fab antibody, and a pharmaceutically acceptable carrier. According to the invention, a monoclonal antibody for effectively neutralizing tetanus exotoxin is screened with natural tetanus exotoxin, and an Fab gene engineering antibody which is produced by large scale in vitro and which has toxin neutralizing effect is prepared with a gene engineering antibody technology on the basis of the monoclonal antibody.

Modified bacterial surface layer (S-layer) proteins are disclosed where the modification is the insertion, at an internal location, of a heterologous polypeptide, or polypeptide of interest. The polypeptide is a binding or target protein, such as an antigen or antibody, or part thereof, in particular a bacterial antigen (e.g. from Clostridium tetani such as TTFC). The modified surface layer protein can then be expressed on the surface of the bacterial cell and used in a vaccine. Also disclosed are bacteria which have been modified to express a heterologous surface layer protein, but which do not as a wild-type possess an S-layer (such as L. casei), and modified bacteria which express only a modified surface layer protein (and not the wild-type S-layer protein). The wild type S-layer is completely replaced with the modified version where the polynucleotide encoding the modified version is integrated into the bacterial genome. The modified S-proteins can form crystalline arrays, sheets or layers that can be used to bind functional molecules (e.g. receptors) to solid surfaces (Au, silicon wafers) in biosensors.

The invention relates to a method for a tetanus toxin receptor binding domain Hc to be subjected to high-level soluble expression in Escherichia coli through nucleotide sequence optimization. According to the sequencing result of a domestic C.Tetani virulent strain CMCC64008, the tetanus toxin receptor binding domain Hc sequence is analyzed and optimized, the optimized sequence is SEQ ID No.1 and the coded protein sequence is SEQ ID No.2. The synthesized Hc gene is linked into an expression vector pET32a(+) after undergoing double enzyme digestion, the recombinant Hc is subjected to high soluble expression in Escherichia coli and the target protein accounts for about 46% of the total protein in the supernatant undergoing bacteriociasis. After QFF column purification, phenyl hydrophobic column purification and SP column purification, the purity of the target protein can be more than 95% and the yield thereof is more than 300mg / L. The recombinant protein prepared by the method of the invention has good immunogenicity, can induce the mice to produce high-titre protective antibodies and can resist attack of high-dose lethal toxins. The method has extensive application prospect in large-scale high-level preparation of the tetanus toxin recombinant subunit vaccine Hc.

The present invention provides a traditional Chinese medicine preparation for treating tetanus, made from cow-bezoar, periostracum cicadae, gastrodia tuber, pearl, notopterygium root, musk, uncaria, frankincense, red peony root, alum, myrrh, poria with hostwood, centipede, borneol, dahurian patrinia, pseudo-ginseng, Ligusticum wallichii, dragons blood, safflower and zhangdan in certain proportioning by weight. The product of the invention can effectively prevent and treat the tetanus, has the function of fever reduction, abirritation, convulsions stopping and convulsion and spasm relieving, has obvious inhibition effect to clostridium tetani, is low in cost, and is safe and effective on the treatment of the tetanus with 95% of effective rate and 89% of curative ratio.

The present invention relates to an improved process of conjugation to obtain synthetic oligosaccharide-protein (OS-PR) conjugates. The process of synthetic OS-PR conjugation is a rapid process providing oligosaccharide-protein conjugates which are highly immunogenic and elicit specific and homogenous immune responses. The synthetic oligosaccharide comprising of four to eight repeating units of respective monomers and at least one in-built terminal amino linker, said synthetic polysaccharide mimics natural polysaccharide obtained from gram negative bacteria such as Neisseria meningitidis serogroups A, C, Y, W, X and Haemophilus influenzae and carrier protein is obtained from gram positive bacteria such as Clostridium tetani (tetanus toxoid) or Corynebacterium diphtheriae (CRM197) or their recombinant versions. The conjugation chemistry of the said oligosaccharide-protein conjugate of the present invention is thio-ether linkage. The present invention takes complete process time in the range of 14-22 hours. The said oligosaccharide-protein conjugates are useful in production of monovalent vaccine or multivalent combination vaccines and as diagnostic tool.

The invention relates to a mouse hybridoma cell strain secreting a clostridium tetani bacillus exotoxin monoclonal antibody, wherein the strain has a preservation number of CCTCC (China Center for Type Culture Collection) NO. C201257. The invention further relates to a clostridium tetani bacillus exotoxin monoclonal antibody prepared by the mouse hybridoma cell strain; the invention further provides an Fab antibody, comprising a kappa chain and an Fd chain; the kappa chain and the Fd chain are obtained by amplifying from total RNA (ribonucleic acid) of the hybridoma cell strain. The invention further relates to a medicine for preventing or treating clostridium tetani bacillus infection, comprising the clostridium tetani bacillus exotoxin monoclonal antibody and / or the Fab antibody, and a pharmaceutically acceptable carrier. According to the invention, a monoclonal antibody for effectively neutralizing tetanus exotoxin is screened with natural tetanus exotoxin, and an Fab gene engineering antibody which is produced by large scale in vitro and which has toxin neutralizing effect is prepared with a gene engineering antibody technology on the basis of the monoclonal antibody.

The invention relates to non-toxic tetanus toxin and clostridium novyi alpha toxin recombinant fusion protein. The prepared recombinant fusion protein is produced in the manner that The prepared recombinant fusion protein is produced in the manner that through codon optimization, a tetanus toxin C fragment, C terminal of clostridium novyi alpha toxin, and N-end non-toxic epitope of clostridium novyi alpha toxin are subjected to fusing expression, so that the immunogenicity of two kinds of toxin protein can be reserved to the maximum extent, and biology potential safety hazard of natural toxincan be avoided. The recombinant fusion protein can be used for preparing clostridium tetani and clostridium novyi subunit vaccines. Compared with the clostridium tetani and clostridium novyi subunit vaccines commercialized in China at present, the non-toxic tetanus toxin and clostridium novyi alpha toxin recombinant fusion protein has the advantages of being simpler in preparation technology, lower in immunizing dosage, better in vaccine effects and the like, the biology security risk in the production process of the vaccine is greatly reduced, and the non-toxic tetanus toxin and clostridium novyi alpha toxin recombinant fusion protein is an ideal candidate vaccine antigen for upgrading and regenerating two clostridial toxin vaccines. When the non-toxic tetanus toxin and clostridium novyialpha toxin recombinant fusion protein and other antigens are in jointed preparation of a combined vaccine, the using dosage of the combined vaccine does not need to be increased, and the combined vaccine can be prepared.

The present invention relates to an improved process of conjugation to obtain synthetic oligosaccharide-protein (OS-PR) conjugates. The process of synthetic OS-PR conjugation is a rapid process providing oligosaccharide-protein conjugates which are highly immunogenic and elicit specific and homogenous immune responses. The synthetic oligosaccharide comprising of four to eight repeating units of respective monomers and at least one in-built terminal amino linker, said synthetic polysaccharide mimics natural polysaccharide obtained from gram negative bacteria such as Neisseria meningitidis serogroups A, C, Y, W, X and Haemophilus influenzae and carrier protein is obtained from gram positive bacteria such as Clostridium tetani (tetanus toxoid) or Corynebacterium diphtheriae (CRM197) or their recombinant versions. The conjugation chemistry of the said oligosaccharide-protein conjugate of the present invention is thio-ether linkage. The present invention takes complete process time in the range of 14-22 hours. The said oligosaccharide-protein conjugates are useful in production of monovalent vaccine or multivalent combination vaccines and as diagnostic tool.

The present invention relates to a pharmaceutical vaccine composition comprising: (a) a pathogen-derived antigen selected from the group consisting of Mycobacterium tuberculosis antigen, Bacillus anthracis antigen, HAV (hepatitis A virus) antigen, HBV (hepatitis B virus) antigen, HCV (hepatitis C virus) antigen, HIV (human immunodeficiency virus) antigen, influenza virus antigen, HSV (herpes simplex virus) antigen, Hib (Haemophilus influenzae type b) antigen, Neisseria meningitidis antigen, Corynebacterium diphtheriae antigen, Bordetella pertussis antigen, Clostridium tetani antigen and Varicella virus antigen; (b) a deacylated non-toxic LOS (lipooligosaccharide); and (c) a pharmaceutically acceptable carrier.

The invention relates to non-toxic tetanus toxin and clostridium perfringens beta toxin recombined fusion protein. By means of codon optimization, clostridium tetani fragments C and the recombined fusion protein containing multiple clostridium perfringens beta toxin mutants with amino acid mutation, the immunogenicity of two kinds of toxin protein is reserved to the maximum extent, and potential biosafety hazards caused by single amino acid mutation are avoided. The recombined fusion protein can be used for preparing unit vaccines of clostridium tetani, B type clostridium perfringens and C type clostridium perfringens; compared with a current commercial clostridium natural toxin inactivated vaccine in China, the recombined fusion protein has the advantages of being simple in preparation process, low in immunizing dose, excellent in vaccine potency and the like, the biosafety risk in the vaccine production process are greatly reduced, and the recombined fusion protein is an ideal candidate vaccine antigen for upgrading and updating the three kinds of clostridium toxin vaccines. When the recombined fusion protein is used for preparing a combined vaccine with other antigens, the combined vaccine can be prepared without increasing the use dose of the combined vaccine.

The invention relates to the field of molecular biology, and particularly discloses a method and a kit thereof for detecting clostridium tetani by using a loop-mediated isothermal amplification method. The method comprises the following steps of: amplifying clostridium tetani spasm toxin genes in a sample DNA by using loop-mediated isothermal amplification technology and using an upstream inner primer, a downstream inner primer, an upstream outer primer and a downstream outer primer, wherein the nucleotide sequences of the primers are expressed as SEQ ID No: 1-4; and detecting the amplification results. The kit comprises the upstream inner primer, the downstream inner primer, the upstream outer primer, the downstream outer primer, DNA quick extraction liquid and isothermal amplification liquid. The method and the kit thereof for detecting the clostridium tetani by using the loop-mediated isothermal amplification method have the advantages of high clostridium tetani specificity, high sensitivity and simple and quick detection process.

The invention discloses a method for preparing a tetanus toxoid vaccine. According to the process, with clostridium tetani strains as raw materials, the tetanus toxoid vaccine is prepared through the following steps of: culturing of tetanus toxoid, bacterium liquid separation, ultrafiltration and concentration, salting out, ultrafiltration desalting and the like. According to the method, firstly culture liquid is subjected to virus-free treatment and then refined, so that the porous channel plugging caused by accumulation of thalli and other impurity segments at a plate and frame membrane package during plate and frame filtering to remove thalli is reduced, and the smoothness during filtration is increased; toxoid protein and other allergens in the culture liquid are removed by changing the salting-out method; and as the desalting methods of the culture liquid after concentration and salting out adopt the tangential flow ultrafiltration method, the destruction of antigen caused by shearing of toxoid protein is reduced, and the protein precipitation is avoided. By utilizing the method, the time for preparing the tetanus toxoid vaccine is shortened, and the production efficiency is improved.