15results about How to "High titer" patented technology

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.

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 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 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 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 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 application belongs to the technical field of bioengineering, and discloses a screening platform for optimizing the interval of baculovirus bidirectional promoters, a screening method and application. The application constructs a high-resolution screening platform with a reference gene integrated in a virus skeleton, and through two-stage screening, first, a transcription enhancement interval is determined by rough screening in the 98-245bp interval, and then, fine screening is performed in the transcription enhancement interval by 1bp steps, so that the optimal interval length of the p10 and polh promoters is determined as 153bp by normalizing the reference fluorescent signal to correct errors such as infection efficiency. The interval is applied to a recombinant adeno-associated virus (rAAV) packaging system, transcription interference between the bidirectional promoters is eliminated, the expression balance of Rep and Cap proteins is restored, the rAAV genome replication efficiency is improved by 3 times, the empty shell rate is reduced from 84.9% to 20.1%, and the transduction efficiency in mammalian cells can be maintained.

The application provides a composition, a method and an application for improving the titer of mouse leukemia virus, and belongs to the technical field of biotechnology.The method comprises the following steps: before a host cell is infected with xenotropic mouse leukemia virus, the host cell is pretreated by using SC79; during virus infection of the pretreated host cell and during cell passage, mifepristone is added to a culture medium; and during continuous virus harvesting, LR3 IGF-1 is added to the culture medium.Through adding additives with different functional mechanisms at different stages of virus production, the application realizes dynamic optimization of the whole process of virus production.The experimental results show that the combination of SC79, mifepristone and LR3 IGF-1 can increase the titer of xenotropic mouse leukemia virus by 14.32 times, which is significantly better than the effect of single additive or two additives in combination.The application solves the problems of low titer of virus stock solution and low production efficiency in the prior art.

This invention discloses the application of β-Hordothionin, its active fragment, or derivatives as intramolecular adjuvants, specifically involving a polypeptide derived from β-Hordothionin, the amino acid sequence of which is shown in SEQ ID NO.1, and the polynucleotide sequence of which is shown in SEQ ID NO.2. The polypeptide is fused with the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein and expressed, significantly enhancing the immunogenicity of the antigen. Experiments show that immunization of mice with the RBD-β-Hordothionin fusion protein induces a 1000-fold increase in specific IgG antibody titers compared to the RBD antigen alone. This fusion protein can be industrially prepared and can be used to enhance the immunogenicity of vaccine antigens.

This invention relates to a genetically engineered strain for synthesizing retinal, its construction method, and its applications. The construction method involves introducing DNA fragments of the β-carotene 15,15'-oxygenase gene, isopentenyl pyrophosphate isomerase gene, phytoene dehydrogenase gene, geranyl-geranyl pyrophosphate synthase gene, and phytoene synthase gene into *Rhizopus cylindrica* to obtain a genetically engineered strain. This strain enhances the expression of β-carotene 15,15'-oxygenase, isopentenyl pyrophosphate isomerase, phytoene dehydrogenase, geranyl-geranyl pyrophosphate synthase, and phytoene synthase in *Rhizopus cylindrica*. By selecting *Rhizopus cylindrica* as a host, introducing key synthetic genes, and achieving stable genome integration, this method yields an engineered strain capable of efficiently synthesizing retinal using glucose as a carbon source, simplifying the production process, reducing costs, and improving industrial applicability.

The present invention relates generally to the field of medicine. More particularly, it concerns the use of pharmacological molecules in the induction of downstream transgene expression from novel inducible cellular promoters.

The invention discloses a method for preparing an influenza virus vaccine by using Vero cells, and belongs to the technical field of biology. The method comprises the following steps: firstly, carrying out serum-free culture amplification on Vero cells in a bioreactor containing a microcarrier; inoculating an influenza virus working seed batch, and maintaining culture in a serum-free culture medium; carrying out three-stage clarification and ultrafiltration concentration on the harvested virus liquid; purifying the virus through sucrose density gradient centrifugation and molecular sieve chromatography; and finally cracking the virus, quantifying, and adding polymer microspheres to prepare a monovalent virus stock solution. The serum-free culture medium comprises pea protein hydrolysate, an amino acid mixture and a cell homeostasis regulating agent. The risk of exogenous animal source components is avoided, the virus harvesting titer is remarkably improved, meanwhile, the immunogenicity of hemagglutinin and neuraminidase is completely reserved, technological parameters are controllable, amplification is easy, and the method is suitable for industrial production.

This invention relates to a culture medium for high-density serum-free suspension culture of BHK-21 cells and its application. The culture medium contains inorganic salts, amino acids, vitamins, energy substances, buffers, antioxidants, sterols, polyamines, additives, and indicators. The antioxidants include dihydrolipoic acid (DLA) and S-acetyl-L-glutathione. This invention provides a culture medium with clearly defined components and significant culture effects for serum-free suspension culture of BHK-21 cells. The added DLA and S-acetyl-L-glutathione, as a combination of antioxidants in the culture medium, enhance the antioxidant properties of the serum-free suspension medium while also improving the cell viability of BHK-21 cells cultured in the medium.