14 results about "Post immunization" patented technology

The present application relates to the technical field of veterinary biological products, in particular to application of oridonin in preparation of poultry vaccine adjuvant. The poultry vaccine adjuvant is composed of oridonin, solvent and buffer. Experiments prove that after mixing the poultry vaccine adjuvant with poultry inactivated vaccine (such as Newcastle disease vaccine) for immunization, the humoral immune level of poultry can be significantly improved (HI titer of 21d after the second immunization is increased by more than 1.5 log2), and the secretion level of specific IgY (systemic immunity) and IgA (mucosal immunity) in serum can be extremely significantly up-regulated, and the high level of antibody is maintained for more than 4 weeks longer than that of the conventional vaccine group. Compared with commercial adjuvant ISA 201, the present application has better immune enhancement activity, high safety, no adverse reaction at injection site, and wide clinical application prospect.

This application discloses a recombinant hemagglutinin protein, its uses, expression methods, and subunit vaccines, belonging to the field of biomedical technology. The technical solution is as follows: a recombinant hemagglutinin protein, obtained by removing the transmembrane and intracellular regions of the HA protein sequence of the H7N9-235 strain and fusing a T4-foldon sequence at the C-terminus; or by retaining the full-length sequence of the HA protein of the H7N9-235 strain. The amino acid sequence of the recombinant hemagglutinin protein is shown in SEQ ID NO.1 or SEQ ID NO.2. The application also discloses a recombinant hemagglutinin protein with good immunogenicity and medical prospects based on the H7N9 strain. Furthermore, when a subunit vaccine is prepared using the recombinant hemagglutinin protein with the transmembrane and intracellular regions removed, the immunogenicity after secondary immunization is significantly enhanced.

The purpose of the present invention is to provide, as a platform technique that may serve as preparation for pandemics, a rapid antibody production technique in which a non-human animal having a human antibody gene is used. More specifically, the purpose of the present invention is to provide an immunization method with which it is possible to induce, in a short period of time, a high antigen-binding ability of an antibody or a high antibody titer of an antiserum, and a method for producing a human antibody / antiserum with which it is possible to rapidly produce a wide range of neutralizing antibodies.　(1) An immunization method for rapidly inducing a human monoclonal antibody having a high ability to bind to a desired antigen or a human polyclonal antibody having a high antibody titer, the method comprising a step for immunizing a non-human animal a plurality of times with a desired antigen or a nucleic acid encoding the antigen within 30 days from the first immunization, the non-human animal having a human antibody gene or locus, and the non-human animal being such that an endogenous gene or locus of the non-human animal corresponding to the human antibody gene or locus is disrupted or deleted, or mutated so as to result in a loss of expression or low expression. (2) A method for producing a human antibody against a desired antigen using a non-human animal, the method comprising: an immunization step for immunizing the non-human animal with the antigen or a nucleic acid encoding the antigen; a human immunoglobulin-positive B cell isolation step for isolating B cells that are human immunoglobulin-positive from a tissue of the immunized non-human animal; an antibody base sequence acquisition step for acquiring the base sequences of antibody light-chain mRNA-derived cDNA and antibody heavy-chain mRNA-derived cDNA prepared from the isolated B cells; and an antibody production step for producing an antibody on the basis of the acquired antibody base sequence, the non-h

This invention discloses the construction of a Toxoplasma gondii progesterone-responsive kinase-deficient strain and its application as a live attenuated vaccine. The Toxoplasma gondii progesterone-responsive kinase-deficient strain is derived from Toxoplasma gondii RHΔ... ku80 As a maternal strain, the gene-editing technology was used to knock out the enzyme encoding progesterone-responsive kinase. prk The gene-deleted strain was named RHΔ ku80 Δ prk Experiments have shown that RHΔ ku80 Δ prk The virulence of the strain was significantly reduced, and immunized mice showed good immunoprotection against acute infection with highly virulent and moderately virulent Toxoplasma gondii strains, and significantly inhibited brain cyst formation in a chronic infection model. Therefore, RHΔ ku80 Δ prk This invention has potential application value as a live attenuated vaccine against Toxoplasma gondii. It has significant application prospects.

The application provides a hybridoma cell strain secreting a monoclonal antibody against a alginate mannuronic acid tetrasaccharide epitope, a monoclonal antibody and application. In order to enhance the immunogenicity of the mannuronic acid tetrasaccharide, a glycoconjugate KLH-1 is used as an immunogen to inject and immunize a mouse, so that the mouse is stimulated to produce a specific immune response against the mannuronic acid tetrasaccharide epitope, then spleen cells of the immunized mouse are fused with myeloma cells, and after screening, an initial hybridoma cell strain is obtained; then after subcloning and specific screening, a hybridoma cell strain capable of stably secreting a monoclonal antibody of the target mannuronic acid tetrasaccharide epitope in alginate is obtained, and a monoclonal ascites antibody is further obtained. The monoclonal ascites antibody shows specific recognition and combination ability for pseudomonas aeruginosa, and the combination activity is related to the expression level of alginate on the surface of the bacteria. Therefore, the monoclonal ascites antibody can be used as a precise detection and diagnosis tool for pseudomonas aeruginosa, and can also be applied to the antibacterial treatment of pseudomonas aeruginosa infection.

This invention relates to the field of bioengineering technology, specifically to a method for preparing neutrophil gelatinase-associated lipid transport protein and its antibody. Specifically, using pET23a(+) as the expression vector, the NGAL gene with six His tags fused to its N-terminus under promoter control is introduced into *E. coli* for induced expression. After purification using a Ni Focurose 6FF column and a molecular sieve column, tag-free NGAL protein with a purity ≥95% is obtained. Subsequently, the NGAL protein is used to repeatedly immunize Japanese rabbits, and serum is collected after each immunization. Qualified serum is screened using a two-dimensional agar diffusion assay, and then purified using the aforementioned antigen to prepare an antigen affinity purification column, yielding a highly active NGAL polyclonal antibody. The high-purity recombinant NGAL of this invention not only possesses activity but is also low-cost, not limited by raw materials, and can be mass-produced, possessing potential and wide-ranging application value in clinical and in vitro diagnostics.

This invention provides a nanobody targeting carp herpesvirus type II and its application. The amino acid sequence of the nanobody is shown in SEQ ID NO. 1-6. The method includes immunizing alpacas with inactivated carp herpesvirus type II, collecting peripheral blood from the immunized alpacas and separating lymphocytes, and extracting total RNA; synthesizing cDNA by reverse transcription, and amplifying the gene fragment encoding the nanobody; ligating it into the pComb3xss vector, and then transforming it into competent cells to construct a nanobody phage display library; enriching and panning the inactivated virus as a coating antigen to obtain specifically binding phage particles; transforming the selected positive clones into host bacteria for induced expression; and obtaining the anti-carp herpesvirus type II nanobody after separation and purification. This invention aims to provide a carp herpesvirus type II nanobody that is low in expression cost and difficulty, has a stable source, can effectively bind to carp herpesvirus type II, and specifically achieves immunoassay of carp herpesvirus type II.

The present application relates to a kind of echinococcus Eg95 protein, nucleotide and mRNA for coding the protein of the present application.By optimizing echinococcus Eg95 protein amino acid and nucleotide sequence, echinococcus circular RNA vaccine, echinococcus linear mRNA vaccine and echinococcus self-replicating mRNA vaccine are prepared, the vaccine can provide strong immune protection to 4-month-old lambs only one needle immunization, so that it can still obtain more than 80% protection (higher than 70% required by Ministry of Agriculture and Rural Affairs) on the 180th day after immunization, solve the problem that commercial sheep echinococcosis genetic engineering subunit vaccine immunity is weak, and frequent vaccination is needed to obtain immune protection, reduce the repeated burden of vaccine procurement and labor cost, reduce the stress response to sheep flock, improve vaccination coverage and compliance, and be conducive to the prevention and control of our country hydatid disease.

The application discloses a kind of liposome polypeptide vaccine of pulmonary inhalation immunity and its preparation method and application, belong to biological medicine technical field.The vaccine includes the liposome of DSPC and DDAB, polypeptide antigen and molecular adjuvant loaded therein.The optimal preparation process condition is: the mass ratio of DSPC and DDAB 5:1, ultrasonic power 275 W, ultrasonic number 50 times, the liposome particle size obtained is about 100 nm, surface is positive charge, meet the physical property requirement of respiratory mucosa immunity.The vaccine can be widely distributed in lung and long time retention after pulmonary mucosa route immunity, promote the uptake and activation of dendritic cell in lung to antigen, further activate T, B cell, produce mucosal immune response, and be taken up after dendritic cell, transport to lymph node, stimulate systemic adaptive immune response.The application provides effective mucosal immune vaccine platform for the prevention and treatment of respiratory pathogenic microorganism infection.

This invention relates to the field of biotechnology and provides a method for screening positive single B cells that secrete target substances and a method for preparing monoclonal antibodies. The positive single B cell screening method includes the following steps: isolating PBMCs from the peripheral blood of immunized animals; sorting memory B cells using magnetic beads and separation antibodies; activating and culturing memory B cells in vitro using an activation medium; and loading, analyzing, and exporting target positive cells from the activated memory B cells using a Beacon single-cell photoconductive system. This invention achieves high-throughput screening of FYL positive single B cells through the Beacon single-cell photoconductive system. Gene products are obtained through reverse transcription and PCR amplification, and accurate sequences are obtained through sequence analysis. The synthesized plasmid is expressed in a eukaryotic system, and high-purity FYL rabbit monoclonal antibodies are obtained through affinity chromatography, laying the foundation for the development of a rapid fentanyl detection kit.

This invention discloses an animal feed containing β-glucan and its preparation method, belonging to the field of animal feed technology. This invention utilizes β-glucan to activate innate immunity, post-biotics to train the immune system, and spirulina to provide immunopolysaccharides and phycocyanin. These three components together form a three-dimensional immune-enhancing network, synergistically strengthening immune function. Furthermore, β-glucan can also function as a prebiotic, inhibiting pathogens and strengthening the immune barrier through the addition of post-biotics, while barley grass powder provides dietary fiber, collectively maintaining intestinal flora balance, optimizing intestinal health, and maintaining the integrity of the physical barrier. Based on improved immunity and intestinal health, combined with balanced nutrition, this improves animal growth performance, especially during stressful periods such as weaning, transportation, and feed changes, promoting growth and stress resistance.

This invention provides a mutant S protein of porcine transmissible gastroenteritis virus (TGEV), a subunit vaccine, and its application; the amino acid sequence of the S protein mutant is shown in SEQ ID NO.3. The S protein mutant of TGEV constructed by this invention is significantly superior to the wild-type SWT protein in both humoral and cellular immunity, and its protein expression level also shows a clear advantage, demonstrating excellent prospects for subunit vaccine development and application. Specifically, in terms of immunity, after the S protein mutant is formulated into a vaccine, this invention can enhance post-immunization antibody levels, TGEV virus neutralizing antibody titers, cellular immune responses, and T lymphocyte proliferation.

Provided in the present disclosure are a Zika / dengue vaccine and its application thereof. The present disclosure introduces a mutation into the E-protein FL fusion region of the Zika virus or dengue virus. Antigens with said mutations are unable to bind to antibodies that causes ADE. After immunization with the vaccine of the present disclosure acquired from the said antigens, production of FL epitope-induced antibodies can be prevented, thereby reducing or eliminating the ADE effect.

The application discloses a specific antibody targeting a lysine K1106 site of an ITSN1-S protein lactylation and a preparation method and application thereof, and the preparation method comprises the following steps: mixing and emulsifying an ITSN1-S-K1106 lactylation antigen with Freund's complete adjuvant and Freund's incomplete adjuvant respectively to obtain a first immunization liquid drop and a booster immunization liquid drop, performing animal immunization by using the first immunization liquid drop and the booster immunization liquid drop respectively, and separating the specific antibody targeting the lysine K1106 site of the ITSN1-S protein lactylation from blood of the immunized animal.The antibody provided by the application can specifically detect the lactylation modification condition of the human ITSN1-S protein from the cell and tissue levels in actual application, and is helpful to explore the action mechanism of post-translational modification of the ITSN1-S protein in tumor proliferation, migration and the like, and provides a potential action target for clinical diagnosis and treatment of tumor diseases.