70 results about "Cholera Toxin B Subunit" patented technology

This invention relates in part to synthesizing high value pharmaceutical proteins in transgenic plants by chloroplast expression for pharmaceutical protein production. We use poly(GVGVP), for example, as a fusion protein to enable hyper-expression of insulin and to accomplish rapid one step purification of fusion peptides utilizing the inverse temperature transition properties of this polymer. We also use insulin-CTB fusion protein in chloroplasts of nicotine free edible tobacco (LAMD 605) for oral delivery. This invention includes expression of native cholera toxin B subunit gene as oligomers in transgenic tobacco chloroplasts which may be utilized in connection with large-scale production of purified CTB, as well as an edible vaccine if expressed in an edible plant, as a transmucosal carrier of peptides to which it is fused to enhance mucosal immunity, and/or to induce oral tolerance of the products of these peptides. The present invention also relates in part to recombinant DNA vectors for enhanced expression of human serum albumin, insulin-like growth factor I, and interferon-α 2 and 5, via chloroplast genomes.

The invention provides a helicobacter pylori multiepitope vaccine. The active ingredient of the helicobacter pylori multiepitope vaccine is a piece of polypeptide; and the helicobacter pylori multiepitope vaccine mainly comprises a multi-copy body of Th and B cell antigen epitopes of helicobacter pylori urease A and B bi-subunit and a mucosal immune adjuvant of a cholera toxin B subunit. The preparation method mainly comprises the following steps of: synthesizing an artificial gene by using a gene synthesis technology, wherein the artificial gene comprises a gene sequence of the multi-copy body of the Th and B cell antigen epitopes of the helicobacter pylori urease A and B bi-subunit; coupling the artificial gene with the gene sequence of the cholera toxin B subunit to form a fusion gene;and expressing the fusion gene by using an escherichia coli prokaryotic expression system, and performing protein purification to obtain the helicobacter pylori multiepitope vaccine. The helicobacterpylori multiepitope vaccine can induce an organism to generate T cell immune response and high-titred specific antibody humoral immune response aiming at the urease A and B bi-subunit, and can be used for preventing and treating helicobacter pylori infection related diseases.

The invention discloses a multimeric protein having the effect of brain targeting, and a preparation method and usage thereof. The method comprises the steps: firstly, expressing cholera toxin B subunit and a fusion protein EGFP-CTA2-TAT of three proteins: an enhanced green fluorescent protein, a cholera toxin A subunit and cell-penetrating peptide in escherichia coli by incompatible double plasmid systems to obtain CTB gene by PCR amplification; cloning the gene segment into a carrier pET-28a to obtain a recombinant plasmid pET-28a-CTB; using wild type CTA2, EGFP and TAT amino acid sequences as templates, inserting 3 enzyme cutting sites and linkers between EGFP and CTA2, and optimizing to obtain codons suitable for expression in escherichia coli; cloning the gene segment into a carrier PET-22b (+) to obtain recombinant plasmid PET-22b-EGFP-CTA2-TAT; using different resistances of PET-28a-CTB and PET-22b-EGFP-CTA2-TAT, and co-transforming the different resistances of the PET-28a-CTB and the PET-22b-EGFP-CTA2-TAT into escherichia coli BL21, to obtain engineering bacteria after screening under double resistance selection pressure of penbritin and kanamycin. CTB5/EGFP-CTA2-TAT chimeric proteins can be obtained after inducible expression of the engineering bacteria by IPTG. The invention further discloses the preparation method and usage of the protein.

The invention discloses a meningococcus capsular polysaccharide polyvalent multivalent conjugate vaccine, a preparation method and application thereof. The polyvalent multivalent conjugate vaccine comprises at least two conjugates of group A, group C, group Y or Group W135 meningococcus capsular polysaccharide and cholera toxin B-subunit. The invention improves the method for extracting the meningococcus capsular polysaccharide, and improves the yield and purity of the capsular polysaccharide by adopting a flocculation extraction method under an electric field.

The invention relates to preparation and application of a mycoplasma hyopneumoniae multi-epitope mucosal vaccine. A mycoplasma hyopneumoniae membrane protein, an adhesive protein P97, a lipoprotein P65, a specific membrane protein P46, a B cell epitope, a Th epitope, a CTL epitope and a cholera toxin subunit B are taken as a vaccine frame structure, a pRSETA carrier is cloned in through flexible linker connection, then Escherichia coli is transformed, and fermentation, purification and preparation technologies are carried out, so that the mycoplasma hyopneumoniae multi-epitope mucosal vaccine with ideal immunogenicity is obtained. A self-made mucosal adjuvant is used in a preparation process, so that production and using processes of the vaccine are simpler and more convenient. Animal experiments show that the mycoplasma hyopneumoniae multi-epitope mucosal vaccine not only has good safety but also can stimulate effective mucosal immunity, humoral immunity and cellular immune reactions.

The invention discloses a vaccine for blocking transmission of echinococcosis pathogeny echinococcus granulosus from a source. The vaccine provided by the invention is a CTB-EgM123 vaccine or a complete set of vaccine consisting of the CTB-EgM123 vaccine and a subunit vaccine of an EgM123 protein recombinant mycobacterium smegmatis vaccine. An active ingredient of the CTB-EgM123 vaccine is a fusion protein formed by fusion of a cholera toxin B subunit and an echinococcus granulosus EgM123 protein; the active ingredient of the EgM123 protein recombinant mycobacterium smegmatis vaccine can express the recombinant mycobacterium smegmatis of the echinococcus granulosus EgM123 protein. The vaccine provided by the invention can stimulate the immune response in a host, improves the immune level of the host, enhances the intestinal mucosal immune response, and plays an important protective role in resisting infection of echinococcosis.

The invention discloses transformant. The transformant comprises a recipient bacterium and a recombinant vector transferred to the recipient bacterium, wherein the recipient bacterium is lactococcus lactis NZ9800; the original vector of the recombinant vector is pNZ8112; and the exogenous target gene of the recombinant vector comprises a porcine somatostatin gene, a cholera toxin B subunit gene and a cell wall anchor stator M6 gene. The transformant can enter the porcine intestinal tract by oral administration; compared with injection immunization, the oral immunization mode reduces the toxicity caused by direct contact with a circulatory system and increases the safety of vaccines; and the immune response of organisms is enhanced and the immune effect is improved by combined immunization of the transformant, an antigen gene and an immune enhancing factor.

A process for preparing cholera toxin B subunit features that by the expression system of Bombyx mori nuclear polyhedrosis virus, the virus with CTB gene is recombined to obtain the recombinant virus (CGMCC No.0594) which has been preserved by Ordinory Microbe Preservation Center of China Microbial Strain Preservation and Management Committee. An oral medicine for treating cholera is prepared through inoculating the virus to the larva or pupa of Bombyx mori, expressing, separating, purifying, and freeze drying. Its advantages are high curative effect, and low cost.

The invention discloses a fusion protein vaccine capable of inhibiting Streptococcus and/or preventing Streptococcus infection. With fusion protein obtained through linkage of sortase A and a cholera toxin B subunit through connecting peptide, the Th17 cell activation level is remarkably increased, the effects of preventing pathogenic bacteria from settling and quickly removing the pathogenic bacteria can be realized, and the fusion protein has protecting effects on Group A Streptococcus of different serotypes and has the superiority of high efficiency, broad spectrum and low cost. With the fusion protein, the use amount of immunogen is reduced, the production technology is simplified, and the production cost is reduced. Meanwhile, the vaccine adopts the way of mucosal immunity, has the characteristics of being free of tissue damage, free of local side effects and convenient to use, and is easy to popularize and use.

The invention relates to an immune modulator for treating arteriosclerosis. Aiming at antigen peptides's weak immunogenicity, a series of epitopes peptides of antigens (SEQ ID NO.1) originating from heat shock protein (HSP60) are inserted into the downstream of Cholera toxin B subunit(CTB), and fusion expression of antigens epitopes peptides and CTB is realized. Produced fusion protein (CTB-p277) and Heat shock protein 65 (HSP65) are coupled chemically, without adjuvant, and directly used immunization. The coupled protein can stimulate body to produce high titer antibody aiming at P277 through mucosal immunity pathway, obviously inhibit occurrence of New Zealand white rabbits's arteriosclerosis, and can have a function for treating arteriosclerosis.

The present invention discloses a preparation method of cholera toxin B subunit protein having biological activity. According to the present invention, a lot of cholera toxin subunits are expressed in Escherichia coli by using a prokaryotic expression vector, and then in vitro efficient renaturation is performed to obtain a large number of the cholera toxin B subunit having biological activity; the operations can be performed in the general laboratory, and no pollution is generated; and the most important advantage is that the labeling with various fluorescence, isotopes and tracers can be performed according to the experimental needs.

The invention discloses an expression vector PVX-6His-CTBt-Bt for producing a multi-epitope vaccine of hepatitis c virus. The multi-epitope vaccine 6His-CTBt-Bt of hepatitis c virus expressed by the vector comprises a histidine tag, an orally-taken immunologic adjuvant cholera toxin B subunit (CTB) and six B cellular antigen epitopes Bp of refractory hepatitis c virus genetype 1. To facilitate high-efficiency expression of the vaccine in tobacco, the 6His-CTBt-Bt gene optimized by an artificially synthesized codon is cloned to a high-efficiency plant virus expression vector potato X virus vector PVX201 to obtain the recombinant expression vector PVX-6His-CTBt-Bt of the multi-epitope vaccine 6His-CTBt-Bt of the hepatitis c virus; the expression vector is inoculated to the tobacco; the tobacco is regarded as a reactor to produce the anti-hepatitis c virus vaccine which aims to the refractory hepatitis c virus, is easy to purify, and can achieve a good orally-taken immune effect and resist against virus infection in vitro.