58results about How to "High titer" patented technology

The invention discloses a recombinant herpes zoster vaccine as well as a preparation method and application thereof, and particularly relates to the technical field of biological medicines. The recombinant herpes zoster vaccine comprises a recombinant herpes zoster vaccine fusion protein purified product HN101 and a liposome mixture HA201, and the recombinant herpes zoster fusion protein gE-gD is constructed by recombining reconstructed gE protein and reconstructed gD protein. The gE-gD recombinant herpes zoster fusion protein is obtained by reconstructing gE protein of varicella-herpes zoster virus and gD protein of HSV through a gene recombination technology and then recombining the gE protein and the gD protein. After the gE-gD recombinant herpes zoster fusion protein is combined with a liposome mixture HA201, stronger immune response can be induced, the antibody titer and cellular immunity are improved, the vaccine effect is enhanced, and meanwhile, the potential infection risk is avoided.

The invention belongs to the technical field of biology, and discloses a method and application for regulating and controlling the fermentation process of schizochytrium limacinum through plant hormones to improve the oil titer, and the method is characterized in that in the fermentation process of schizochytrium limacinum, one or more plant hormones are added into a culture medium to promote accumulation of oil. Plant hormones are added into a fermentation culture medium of schizochytrium limacinum, the plant hormones comprise gibberellin (GB), 6-benayl aminopurine (6-BAP), forchlorfenuron (CPPU) and brassinolide (BR), and the plant hormones promote grease accumulation by adjusting activity of enzymes related to fatty acid synthesis, so that the plant hormones can adjust metabolic pathways of the schizochytrium limacinum and promote grease accumulation. The synthesis efficiency and the yield of the grease are obviously improved.

The invention relates to the technical field of gene engineering, and particularly discloses application of a gene recombinant of an adenovirus vector and a p53 gene, and application of the recombinant constructed by utilizing the adenovirus vector and the human p35 gene in preparation of drugs for treating proliferative diseases. The recombinant adenovirus is packaged by adopting an AdMax packaging system framework, the E1 / E2-deleted replication-deficient adenovirus is obtained through intracellular homologous recombination, recombinant virus particles with relatively high titer can be obtained, and the recombinant virus particles can have relatively high expression in cells with proliferative diseases, so that the recombinant adenovirus can be used for treating proliferative diseases. By means of the method, in-vivo transfer of the exogenous p53 gene and efficient expression of the exogenous p53 gene in hyperplasia tissue of a patient are achieved, so that the purpose of treatment is achieved, and gene treatment of hyperplasia diseases is achieved.

The invention relates to the technical field of biology, and particularly discloses a recombinant vector, a recombinant baculovirus and a preparation method and application of the recombinant vector and the recombinant baculovirus. According to the method for preparing the recombinant vector, pFastBac Dual is adopted as a vector to prepare a recombinant plasmid vector inserted into a VP2 gene of an avian bursal disease virus classic strain, and the recombinant plasmid vector is named as pDual-V1; the method comprises the following steps: preparing a recombinant plasmid vector into which VP2 genes of a classical strain and a variant strain of the avian bursal disease virus are inserted at the same time, and naming the recombinant plasmid vector as pDual-V1-V2; the method comprises the following steps: preparing a recombinant plasmid vector inserted into a group I type 4 fowl adenovirus Fiber-2 gene, and naming the recombinant plasmid vector as pDual-F2; the method comprises the following steps: preparing a recombinant plasmid vector into which a Fiber-2 gene and an avian reovirus sigma C gene sequence are inserted at the same time, wherein the recombinant plasmid vector is named as pDual-F2-sigma C; carrying out double enzyme digestion on the pDual-V1-V2, recovering small fragments, carrying out double enzyme digestion on the pDual-F2-sigma C, recovering large fragments, and then carrying out mixed enzyme ligation. The recombinant baculovirus can express four antigens at the same time, and effective preparation of a multi-combined vaccine is achieved.

The invention relates to the technical field of immune carriers, in particular to immune carrier microspheres loaded with individualized MHC-II binding polypeptide and vaccine preparation and application of the immune carrier microspheres loaded with the individualized MHC-II binding polypeptide. The core microsphere is loaded with individualized MHC-II binding polypeptide, the sequence of the MHC-II binding polypeptide is obtained by predicting and screening based on an HLA genotyping result, and each HLA allele corresponds to at least one high-affinity MHC-II binding polypeptide; and the shell is a glucan or other polymer coating layer. The preparation method has the advantages that the T epitope and the B epitope are separately subjected to immune competitive inhibition inside and outside the microspheres, so that a better immune effect is obtained. The antibody avoids cross reaction side effects; carrier molecule diversity is reduced, and side effects caused by T cell over-activation are avoided; th1 epitopes and Th2 epitopes can be contained in the microspheres, so that antibody immunity and T cell immunity functions are generated; the particle size of the microspheres is controllable, and the immunologic function can be achieved without adjuvants.

Provided are an engineered EB virus nuclear antigen 1 (EBNA1) and a coding molecule thereof, a vector and mammalian cell expression system comprising the EBNA1 and the coding molecule thereof, and a target polypeptide recombined by the aforementioned contents. Also provided are methods of making the engineered EBNA1, and encoding molecules, vectors, and mammalian cell expression systems thereof, and methods of using the same for recombinant expression.

The invention discloses a gene VII type Newcastle disease attenuated marker vaccine strain rMG7-HN-G256E, the preservation number of the gene VII type Newcastle disease attenuated marker vaccine strain is CGMCC (China General Microbiological Culture Collection Center) No.46292, and the whole-genome nucleotide sequence of the vaccine strain is as shown in SEQ ID No: 1; wherein the 256th amino acid glycine (G) of the HN protein is replaced by glutamic acid (E). A Newcastle disease virus reverse genetic manipulation platform is utilized, different sites are mutated on the basis of a Newcastle disease virus MG7 strain, a gene VII type Newcastle disease virus marker vaccine attenuated strain rMG7-HN-G256E which is low in pathogenicity, good in growth characteristic, high in virus titer, stable in passage and excellent in immunogenicity is rescued, and the marker vaccine strain is good in immunogenicity, high in immunogenicity, high in immunogenicity, high in specificity, high in specificity and high in specificity. The vaccine can induce a high-level protective antibody, completely protects immunized chickens, can be used for preventing and controlling the current epidemic gene VII type Newcastle disease virus, and lays a foundation for identifying vaccine immunity and wild virus infection.

The invention discloses a pre-fusion RSV F protein mutant and application thereof. The invention provides a new RSV F protein mutation site, it is found for the first time that 157C and 183C substitution and / or 170C and 177C substitution in the respiratory syncytial virus F protein has an important effect on stabilizing conformation before fusion, S155C and S290C substitution can be not used, the immunogenicity of the respiratory syncytial virus F protein before fusion is improved, the neutralizing antibody titer induced by the respiratory syncytial virus F protein is greatly improved, and the immunogenicity of the respiratory syncytial virus F protein before fusion is improved. Meanwhile, the high expression level of the antigen in mammalian cells is achieved, and the method is used for preparing RSV diagnostic reagents, antibodies and various vaccines with the pre-fusion F protein as the antigen.

The invention mainly provides a method for producing oncolytic viruses by using a bioreactor. Through perfusion culture, the Vero cell quantity during virus infection is optimized, so that the purposes of increasing the virus yield, reducing the batch difference and finally producing the oncolytic virus can be achieved.

The application discloses a phosphoric acid modified protein or polypeptide, a preparation method thereof and application thereof in vaccines, and belongs to the field of protein phosphoric acid modification. The preparation method of the phosphoric acid modified protein or polypeptide is as follows: a compound containing an aldehyde group and a phosphate group is subjected to nucleophilic reaction with a protein or polypeptide to form a Schiff base; a reducing agent is used to reduce the Schiff base, and after post-treatment, the phosphoric acid modified protein or polypeptide is obtained. After the phosphoric acid modified protein product prepared by the method is prepared into a vaccine with an aluminum hydroxide adjuvant and mice are immunized, the antibody titer is significantly increased. This is of great significance for increasing the immune effect, reducing the inoculation dose, saving the antigen amount and improving the immune response of some people or in specific conditions.

The invention discloses preparation and application of a circular RNA (Ribonucleic Acid) molecule of a foot and mouth disease virus. The circular RNA comprises a coding element for coding any one of foot and mouth disease virus VP1 protein and VP3 protein; preferably, the coding element encodes foot and mouth disease virus VP1 protein. Further, the coding element codes a VP1 wild type protein, a VP1 truncated protein, a VP1 mutant protein and a VP1 truncated mutant protein; preferably, the coding element encodes the VP1 truncated and mutant protein. Further, the encoding element encodes an optimized nucleotide sequence of foot-and-mouth disease virus VP1 truncated and mutant protein. Further, the circular RNA comprises an optimized translation initiation element. Researches show that the circular RNA provided by the invention can induce protein expression in an animal body, generate a high-level virus blocking antibody and successfully activate T cell immunity, especially CD8 + T cell immune response, and a basis is provided for preparing a circular mRNA vaccine for preventing and / or treating foot-and-mouth disease.

The invention discloses a conjugate containing a trehalose derivative and a carbohydrate antigen as well as a preparation method and application of the conjugate, and belongs to the technical field of anti-tumor carbohydrate vaccines. The conjugate is obtained by taking an immunologic stimulant trehalose derivative of a mincle receptor as an embedded adjuvant to be conjugated with a sugar antigen TF, the trehalose derivative can improve the immunogenicity of the TF sugar antigen, the TF sugar antigen is presented to a corresponding immune cell, and a higher-titer immune response aiming at tumor sugar antigen TF specificity is generated. In addition, the compound also has the advantages of clear structure, simple and convenient synthesis method, stable and controllable product quality and the like, and can overcome the defect of weak immunogenicity of sugar antigens; meanwhile, under the action of an embedded adjuvant trehalose derivative, the vaccine can also generate immune memory.

The invention discloses preparation and application of a rabies virus circular RNA (Ribonucleic Acid) molecule. The circular RNA comprises a coding element for coding rabies virus G protein. Further, the encoding element encodes a wild type G protein, a G truncated protein, a G mutant protein and a G truncated and mutant protein; preferred coding elements encode G truncated plus muteins. Further, the encoding element encodes an optimized nucleotide sequence of rabies virus G truncated plus mutant protein. Further, the circular RNA comprises an optimized translation initiation element. Research shows that the circular RNA provided by the invention can induce protein expression, a higher-level neutralizing antibody and a more lasting drug effect can be generated by only one-time administration in an animal body, and a basis is provided for preparing a circular mRNA vaccine for preventing and / or treating rabies.

The invention relates to a conjugate of a dual agonist and a carbohydrate antigen, the conjugate contains the dual agonist and the carbohydrate antigen, the dual agonist is an iNKT cell agonist and a mincle receptor agonist, the iNKT cell agonist is an alpha-galactosylceramide analogue, and the mincle receptor agonist is a trehalose derivative; the structural general formula of the conjugate is as shown in formula (I): # imgabs0 # X is a carbohydrate antigen, and Y is a linker. The vaccine prepared from the conjugate can activate iNKT cells and mincle receptors at the same time, induce generation of high-titer IgG antibodies and effectively improve the immunogenicity of carbohydrate antigens, and has a good anti-tumor application prospect.

The present invention relates to a fermentation process for the production of biosurfactants.

The invention belongs to the technical field of biology, and particularly relates to a blue-light-responsive corynebacterium glutamicum engineering strain and a construction method and application thereof.According to the corynebacterium glutamicum engineering strain, corynebacterium glutamicum serves as a host, a light-operated gene is introduced for blue-light response, and the blue-light-responsive corynebacterium glutamicum engineering strain is obtained. The light-operated gene is a BsLicTCAT-VVD fusion protein coding gene which is formed by fusing a DNA (Deoxyribose Nucleic Acid) sequence of an RNA (Ribonucleic Acid) binding structural domain LicTCAT for transcribing an anti-terminating protein LicT and a VVD protein coding gene sequence. Compared with the prior art, the light-controlled gene BsLicTCAT-VVD fusion protein coding gene is introduced to carry out blue light response, the target gene is constructed at the downstream of the RAT anti-terminator, and the expression of the target gene is regulated through a blue light signal, so that the corynebacterium glutamicum has the capability of finely regulating the expression quantity of various genes in cells.

According to the double-plasmid packaging system for improving the AAV2 virus titer, the packaging system is co-transfection of an AAV producing cell and two plasmids, and the two plasmids are shuttle plasmids which are provided with two AAV ITRs, are respectively inserted with FRT sequences on two sides and are loaded with target genes; the invention relates to a helper plasmid which is provided with rep-cap, can express different serotypes, is inserted into a human miRNA (micro Ribonucleic Acid) sequence and a CAG-Flpo-wpre-pa sequence and provides a helper gene separated from an adenovirus. According to the system disclosed by the invention, the AAV2 virus with the titer of 1.0 * 10 < 10 > can be packaged by using a double-plasmid co-transfection system. The difficulty of plasmid cotransfection is reduced, and a product route is provided for commercial production of AAV2 virus packaging.

The invention discloses a chemical synthesis method of a type b haemophilus influenzae capsular polysaccharide derivative, and belongs to the field of carbohydrate chemistry. The invention provides a modular synthesis strategy based on a phosphoramidite method. According to the strategy, a dimer receptor with a connecting arm at the O3 site of ribose at a non-reducing end is used as a main starting compound, and a phosphoramidite monomer donor and a dimer donor are respectively used as extension units of an odd-numbered polymer synthesis route and an even-numbered polymer synthesis route. Through optimization of a coupling catalysis strategy and optimization of a protecting group removal method, chemical synthesis of 1-12 repetitive units is efficiently completed. According to the synthesis method, the raw materials are cheap and easy to obtain, the preparation method is simple and easy to repeat, the yield of each step is relatively good, and the method has a very good application prospect in vaccine development of the haemophilus influenzae type b.

The invention provides a vaccine adjuvant, a protein used by the vaccine adjuvant and a preparation method of the protein. The technical problem to be solved is how to provide an effective and nontoxic vaccine adjuvant. The invention discloses a protein. The protein is any one of the following items: A1) a protein with an amino acid sequence as shown in a sequence 2; a2) a protein which is obtained by substitution and / or deletion and / or addition of amino acid residues of the protein in A1), has 80% or more of identity with the protein in A1) and has the same function as the protein in A1); and A3) a fusion protein obtained by connecting the N terminal or / and C terminal of A1) or A2) with a protein tag. The invention discloses an influenza virus adjuvant which is named as protein Ad-20, and the protein Ad-20 serving as the influenza virus adjuvant can stimulate humoral immunity and cellular immunity, has the functions of stimulating maturation of B cells and promoting the B cells to secrete antibodies in advance, is used for enhancing immunogenicity and immune memory of virus vaccines, is non-toxic and has the effect higher than that of an aluminum adjuvant; and a new auxiliary means is provided for research and development of effective vaccines.

This invention discloses a nucleic acid molecule encoding a modified retroviral Gag protein and its applications. The Gag protein contains an engineered acylated tag sequence at its N-terminus, comprising: a glycine residue at the second position after the initiating methionine for myristylation modification; and / or a cysteine ​​residue in the N-terminal region for palmitoylation modification. The tag sequence is selected from SEQ ID NO:1, 3-9, or derivative amino acid sequences having at least 80% identity and retaining function. Preferably, the modification involves replacing the first 9 amino acids at the N-terminus of the wild-type HIV-1 Gag protein with the aforementioned tag to enhance membrane anchoring and improve viral packaging efficiency. This invention also provides packaging plasmids containing this molecule, host cells, packaging systems, and preparation methods. This invention can significantly improve viral yield and titer, and is particularly suitable for overcoming the efficiency bottleneck in packaging large-load lentiviruses of 7-10 kb.

The invention discloses a gene VII type Newcastle disease attenuated marker vaccine strain rMG7-F-N471A, the preservation number of which is CGMCC (China General Microbiological Culture Collection Center) No.46293. The whole-genome nucleotide sequence of the vaccine strain is as shown in SEQ ID No: 1; wherein the 471st amino acid asparagine (N) of the F protein is replaced with alanine (A). A Newcastle disease virus reverse genetic manipulation platform is utilized, different sites are mutated on the basis of a Newcastle disease virus MG7 strain, a gene VII type Newcastle disease virus marker vaccine attenuated strain rMG7-F-N471A which is low in pathogenicity, good in growth characteristic, high in virus titer, stable in passage and excellent in immunogenicity is rescued, and the marker vaccine strain is good in immunogenicity, high in immunogenicity, high in immunogenicity, high in specificity, high in specificity, high in specificity and high in specificity. The vaccine can induce a high-level protective antibody, completely protects immunized chickens, can be used for preventing and controlling the current epidemic gene VII type Newcastle disease virus, and lays a foundation for identifying vaccine immunity and wild virus infection.

The application discloses a short peptide for TRPM7 channel, a vaccine and pharmaceutical application of the short peptide for preventing and treating atrial fibrillation, and belongs to the technical field of biological medicine. The short peptide can be coupled with a carrier KLH by a glutaraldehyde coupling method to prepare a conjugate, and the conjugate is supplemented with an adjuvant to form a vaccine. The vaccine can induce the body to produce high-specificity and high-titer antibodies through active immunization, the antibodies can block the TRPM7 channel, inhibit calcium influx of atrial muscle cells and atrial fibroblasts, reduce calcium overload of cytoplasm, inhibit atrial electrical remodeling and tissue remodeling, and prevent and treat atrial fibrillation. The application provides a pharmaceutical composition containing the short peptide, the conjugate and the adjuvant, and a polyclonal antibody obtained by immunizing a non-human animal with the vaccine. Compared with existing chemical drugs, the application has the advantages of high targeting, long-acting, good safety, low price and the like, provides a new active immunotherapy strategy for atrial fibrillation patients, and has a wide clinical transformation prospect.

The invention discloses a gene VII type Newcastle disease attenuated marker vaccine strain rMG7-L-V1291I, the preservation number of the gene VII type Newcastle disease attenuated marker vaccine strain rMG7-L-V1291I is CGMCC (China General Microbiological Culture Collection Center) No.46291, and the whole-genome nucleotide sequence of the vaccine strain is as shown in SEQ ID No: 1; wherein the 1291th amino acid valine (V) of the L protein is replaced by isoleucine (I). A Newcastle disease virus reverse genetic manipulation platform is utilized, different sites are mutated on the basis of a Newcastle disease virus MG7 strain, a gene VII type Newcastle disease virus marker vaccine attenuated strain rMG7-L-V1291I which is low in pathogenicity, good in growth characteristic, high in virus titer, stable in passage and excellent in immunogenicity is rescued, and the marker vaccine strain is good in immunogenicity, high in immunogenicity, high in specificity, high in immunogenicity and capable of being used as a marker vaccine for the Newcastle disease virus. The vaccine can induce a high-level protective antibody, completely protects immunized chickens, can be used for preventing and controlling the current epidemic gene VII type Newcastle disease virus, and lays a foundation for identifying vaccine immunity and wild virus infection.

The invention discloses an HEK293 derived cell strain capable of stably expressing adenovirus ADP as well as a construction method and application of the HEK293 derived cell strain. The preservation number of the cell strain is GDMCC No.65235. According to the cell strain, an ADP gene complete ORF of an Ad5E3 region and an expression cassette of the ADP gene complete ORF are stably inserted into an AAVS1 site of HEK293. Compared with an HEK293 cell, the derivative cell strain can promote the formation of replication-deficient Ad5 vector plaques, increase the copy number of the replication-deficient Ad5 vector and improve the specific activity of the replication-deficient Ad5 vector, can be applied to the production of Ad5 vector vaccines and gene therapy products, increase the yield and reduce the industrialization cost, and has transformation application value.

The invention discloses a vaccine liposome, the vaccine liposome comprises a liposome and a vaccine, the liposome comprises phospholipid and cholesterol, and the mass ratio of the liposome to the vaccine is (15-60): 1. The invention relates to a vaccine liposome and a soluble microneedle administration composition added with the liposome, compared with a metal microneedle and the like, the vaccine liposome and the soluble microneedle administration composition added with the liposome are safer and free of residues, active matters of the skin administration composition are in a dissolved or suspension state, and release of drugs / vaccines on epidermis can be promoted; and better, higher and quicker transdermal release effect and in-vivo antibody titer can be realized by using one fifth of active matters in the conventional means compared with the conventional means.

This invention discloses a method for efficiently isolating human parainfluenza virus based on suspended MDCK cells. By using the fully suspended passaged MDCK cell line for human parainfluenza virus isolation and culture and optimizing the inoculation process, single-generation virus amplification can be completed within 3 days, significantly shortening the cycle, improving virus isolation efficiency and success rate compared to traditional adherent culture methods, and enhancing experimental reproducibility. This method not only demonstrates good isolation and culture capabilities for the major subtypes of human parainfluenza virus, but also shows good adaptability to clinical samples from different sources and with different initial loads. In addition, this culture system has linear scale-up potential, is not limited by the surface area of ​​adherent culture, and can meet the large-scale virus seed preparation needs from clinical testing to vaccine mass production, providing key technical support for the integrated process from human parainfluenza virus isolation to vaccine production.

The invention relates to a radiopharmaceutical passive skin anaphylaxis test method, which comprises the following steps: animal grouping: dividing test animals into a positive control group, a negative control group and a drug group, and marking; preparing the medicine: respectively mixing the negative reference substance, the positive reference substance and the radiopharmaceutical with the adjuvant to obtain each group of mixed emulsion; preparing antibody serum: taking a plurality of test animals from each group, performing drug injection, sampling blood on the fourteenth day after the last sensitization, separating the serum, and diluting to obtain antibody serum with medium, high and low concentrations; passive sensitization: taking a plurality of normal test animals from each group, injecting corresponding antibody serum into each group, and carrying out passive sensitization; and excitation: after sensitization is carried out for 48 hours, injecting each group of solutions containing dye liquor into each group of test animals for excitation, measuring the size of blue spots, and determining that the test animals with the diameter greater than 5 mm are positive. The method can be used for safely, effectively, simply and conveniently carrying out preclinical evaluation research on the sensitization of various radiopharmaceuticals.

The present invention belongs to the field of molecular biology technology, and provides the use of GSK3β in enhancing Seneca virus replication for the preparation of vaccines. A method for increasing the titer of Seneca virus in host cells for vaccine preparation is also provided: (1) constructing a host cell overexpressing GSK3β; (2) culturing the host cell overexpressing GSK3β and inoculating Seneca virus. The present invention successfully constructs the expression vector pCMVMyc‑GSK3β of GSK3β by molecular biology technology; and constructs a BHK‑21 cell line overexpressing the GSK3β gene; after inoculation of Seneca virus, the cell line has an accelerated viral replication rate and a significantly increased viral titer, and can be used for the commercial production of Seneca vaccine.

The invention relates to the field of biological pharmacy, in particular to fusion protein, a nano vaccine and application thereof. The fusion protein provided by the invention comprises a fusion protein 1 and / or a fusion protein 2; the fusion protein 1 is prepared from a SpyTag protein and an Rv1498A protein; and the fusion protein 2 comprises a SpyCatcher protein and a gp350 protein. A flavin binding protein Rv1498A in a mycobacterium tuberculosis dodecin-like family is used as a carrier protein, a distal membrane end and a proximal membrane end of an EB virus gp350 extracellular domain are respectively displayed on the surface of the Rv1498A by utilizing a SpyTag-SpyCatcher technology, two immunogenic compounds are obtained, high expression quantity is realized while correct modification and folding of an antigen are ensured, and the EB virus gp350 extracellular domain immunogenic protein Rv1498A can be used for preparing the EB virus gp350 extracellular domain immunogenic protein Rv1498A and the EB virus gp350 extracellular domain Rv1498A immunogenic protein Rv1498A and the EB virus gp350 extracellular domain Rv1498A. And the nano-particle vaccine prepared by the aid of an adjuvant is proved to have better immunogenicity in a mouse body.

The invention relates to a method for improving the expression quantity of viruses, in particular to a method for improving the expression quantity of bovine rotaviruses. The method comprises the following steps: adding Marc-145 cells into a cell culture bottle or a cell culture plate acted by polylysine for subculture, and then inoculating bovine rotavirus for culture. According to the method, the Marc-145 cells are cultured by using the cell culture bottle or the cell culture plate acted by the polylysine, so that the tolerance degree of the Marc-145 cells to high-concentration trypsin is improved, and a proper pancreatin tolerance concentration value is screened out. Furthermore, the bovine rotavirus is inoculated into a DMEM culture medium containing Marc-145 cells and high-concentration trypsin, so that the expression quantity of the bovine rotavirus is improved, and the virus titer of virus expression is remarkably improved.