67 results about "Vaccine research" patented technology

The invention relates to the technical field of biological detection, in particular to a monoclonal antibody combination for HPV18 type E6 protein detection and application. The provided combination is composed of 5G3 and 2C7, and the amino acid sequences of complementary determining regions of variable regions of a heavy chain and a light chain of the combination are clear and are respectively shown as SEQ ID NO.1-12. The antibody combination has high specificity and sensitivity, the lowest detection limit can reach 100 pg / ml, and cross reaction with other HPV subtypes is avoided. According to a double-antibody sandwich ELISA and biotin-avidin amplification detection system constructed based on the combination, the signal intensity and the detection accuracy are remarkably improved, and the combination is suitable for rapid detection of the HPV18 type E6 protein in a cervical exfoliated cell sample and has application value in early diagnosis of cervical cancer, risk stratification, vaccine research and development and curative effect evaluation.

The invention relates to the technical field of biological medicine, in particular to a monoclonal antibody of bovine coronavirus S2 protein, an epitope of the monoclonal antibody and application of the epitope. The invention provides a hybridoma cell strain (preservation number: CCTCC (China Center For Type Culture Collection) NO: C202510). The cell strain can secrete a monoclonal antibody which is targeted and combined with bovine coronavirus S2 protein; the amino acid sequence of the epitope identified by the monoclonal antibody is shown as SEQ ID NO: 1. Experiments prove that the monoclonal antibody has neutralizing activity, and the targeting epitope of the monoclonal antibody is a key neutralizing epitope. The invention provides a new strategy and tool for antibody drug and vaccine research and development as well as diagnosis and detection of the bovine coronavirus.

The invention relates to the technical field of biological medicine, and particularly discloses a monoclonal neutralizing antibody for resisting respiratory syncytial virus and application of the monoclonal neutralizing antibody. The antibody provided by the invention can identify non-denatured RSV (L) protein through ELISA (enzyme-linked immunosorbent assay), but cannot identify denatured RSV (D) protein, and is a kappa IgG2b type antibody. The molecular marker is 0.0001 mu g / ml-1 mu g / ml, can positively recognize the RSV Pre-F protein, and has a gradient effect. The antibody can be used as a neutralizing active antibody in an ELISA double-antibody sandwich method, can be used for detecting the in-vitro potency of the RSV PreF protein, and plays an important role in evaluating the vaccine activity in the vaccine research and development process.

The invention discloses an antigen LARP1 of cysticercus bovis and application of the antigen LARP1, and belongs to the technical field of biochemistry. The invention aims to provide a detection method and a treatment and prevention method for cysticercosis bovis. The invention provides an antigen of cysticercus bovis. The amino acid sequence of the antigen of cysticercus bovis is as shown in SEQ ID NO. 4. And a new candidate antigen is provided for research and development of novel vaccines, development of specific diagnostic reagents and the like.

The invention discloses application of MY-1B in preparation of a medicine for preventing and treating Nipah virus, and belongs to the technical field of medicine. The invention mainly aims at the m5C modification of a host, and further reduces the m5C modification of the host by inhibiting the methyltransferase activity of NSUN2 so as to inhibit the replication of the Nipah virus, and has a better effect. At present, most of research and development of virus drugs and vaccines are designed or modified aiming at viruses, a new thought is provided for research and development of drugs and vaccines in the future, and a new application is also provided for MY-1B.

The invention discloses an attenuated, broad-spectrum and pancreatin-independent porcine epidemic diarrhea virus recombinant strain as well as a construction method and application thereof, and belongs to the technical field of animal virology and genetic engineering. According to the present invention, the 894-993 site amino acid sequence of the PEDV AH2012 / 12 variant strain S protein is transformed into the 894-993 site amino acid sequence of the JS2008 classical strain S protein through the CRISPR / Cas9 system and the homologous recombination method, and the recombinant virus rAH2012 / 12-894-993aaJS2008 is obtained through the rescue; compared with a parent virus AH2012 / 12, the recombinant virus rAH2012 / 12-894-993aaJS2008 has the advantages that the proliferation titer in cells is obviously improved, the toxicity is highly weakened, a cross-protective neutralizing antibody aiming at the G1 type and the G2 type of the PEDV can be induced, and the virus strain can be used for fundamental research and vaccine research and development.

The application discloses a SARS-CoV-2 epitope type vaccine multi-epitope combination and application, and belongs to the technical field of coronavirus vaccine research and development.The first aspect of the application relates to a fusion protein, which comprises in sequence: (a) a SARS-CoV-2 spike protein receptor binding domain or a functional fragment thereof; (b) a T cell epitope domain, comprising: a CTL epitope cluster, the CTL epitope cluster comprising at least one CD8+ T cell epitope polypeptide selected from SEQ ID NO: 1-15; and (c) an immunoglobulin Fc domain.The application adopts a tandem strategy of immunodominant epitopes + conserved epitopes, predicts high-affinity T cell epitopes by computational biology methods, evaluates the HLA restriction in different populations, introduces a flexible linker peptide for optimization design, evaluates the immune effect difference of different combinations through in vitro and animal models, analyzes the synergistic or competitive relationship between epitopes, and optimizes the vaccine design.Through systematic comparison of the immunological effect difference of different epitope combinations and the adaptability to various vaccine platforms, the application establishes an optimized safe, efficient, broad-spectrum and long-acting multi-epitope vaccine design strategy, and has significant application value and important transformation value.

The present application relates to the technical field of biotechnology, in particular to a monoclonal antibody against A29L protein of monkeypox virus and application thereof. The present application prepares the monoclonal antibody against A29L protein of monkeypox virus, and through optimization of antibody screening and expression technology, the antibody with high affinity and high specificity is screened, and then a new tool is provided for rapid detection, effective neutralization and treatment of monkeypox virus, and an important immunological basis is provided for vaccine research and development and treatment scheme design of monkeypox virus.

The invention provides a human diploid stably transfected cell line suitable for coxsackie virus A6 culture and application of the human diploid stably transfected cell line. The construction of the human diploid stably transfected cell line comprises the steps of construction of an hKREMEN1 lentivirus overexpression vector, virus packaging and stably transfected cell screening. The problem that proliferation of the CVA6 virus in human diploid cells is not ideal is solved, the human KREMEN1 gene is over-expressed in the human diploid cells (KBM17) by utilizing a lentivirus technology, and the proliferation capacity of the CVA6 virus on the KMB17 cells is improved by constructing the KMB17 cells capable of stably expressing hKREMEN1 receptors, so that a good foundation is laid for research and development of hand-foot-and-mouth disease vaccines; vaccine preparation and effectiveness evaluation are facilitated.

The invention discloses application of carfilzomib in preparation of a medicine for preventing and treating Nipah virus, and belongs to the technical field of medicine. The method mainly aims at ubiquitination modification of nipah virus M protein, the carfilzomib is used for inhibiting a host proteasome path, free ubiquitination factors in cells are depleted to achieve the effect of inhibiting ubiquitination of the M protein, then nucleoplasm shuttle of the M protein is inhibited, replication of the nipah virus is inhibited, and the effect is good. At present, most of research and development of virus medicines and vaccines are designed or modified aiming at viruses, a new thought is provided for research and development of medicines and vaccines in the future, and a new application is also provided for Carfilzomib.

The invention discloses application of STM2457 in preparation of a medicine for preventing and treating Nipah virus, and belongs to the technical field of medicine. The application mainly aims at m6A modification of a host, the activity of methyltransferase of METTL3 is inhibited through STM2457, then m6A modification of the host is reduced, replication of the Nipah virus is inhibited, and a good effect is achieved. At present, most of research and development of virus medicines and vaccines are designed or modified aiming at viruses, a new thought is provided for research and development of medicines and vaccines in the future, and a new application is also provided for STM2457.

The present invention belongs to the field of biotechnology, and discloses the use of an LMH cell line overexpressing the RPL7 gene, and a vaccine; Infecting goose astrovirus into an LMH cell line capable of overexpressing goose-derived RPL7 can, to a certain extent, promote the proliferation of goose astrovirus. As a component of the large ribosomal subunit, RPL7 directly participates in the process of mRNA to protein synthesis. GAstV mRNA interacts with RPL7 and localizes to the endoplasmic reticulum, greatly promoting the reverse transcription level of the virus, and thus promoting the replication of goose astrovirus; When it is applied to vaccine research and development, it can also obtain goose astrovirus with a high titer, which helps to reduce the cost of vaccine research and development and improve the effect of the vaccine.

The invention discloses an anti-zebra fish IgM monoclonal antibody and preparation and application thereof, and belongs to the technical field of monoclonal antibodies, and the anti-zebra fish IgM monoclonal antibody is secreted by a hybridoma cell strain with the preservation number of CCTCC NO: C202542. The anti-zebrafish IgM monoclonal antibody provided by the invention not only can specifically recognize IgM in serum and IgM + B cells in tissues, but also can be used for evaluating the immune effect of vaccines, is wide in application, and has positive significance on scleroderma immunity and vaccine research.

The invention belongs to the technical field of in-vitro transcription, and particularly discloses a 5 'UTR element with a protein expression enhancing function, an expression vector and application. The 5 'UTR is selected from any one of an SPP1d element, a TSPAN1u1 element and an SPP1d-TSPAN1u1 series element; the nucleotide sequence of the SPP1d element is as shown in SEQ ID NO: 1, the nucleotide sequence of the TSPAN1u1 element is as shown in SEQ ID NO: 2, and the nucleotide sequence of the SPP1d-TSPAN1u1 tandem element is as shown in SEQ ID NO: 3. The 5 'UTR element can significantly improve translation efficiency, has a wide enhancement effect on functional protein expression, is high in biological safety, and provides a standardized and high-performance translation regulation tool for the fields of efficient recombinant protein production, gene therapy vector optimization, vaccine research and development and the like.

The present application relates to the technical field of biological medicine, in particular to a kind of mRNA vaccine for obtaining bovine nodular skin disease virus neutralizing antibody and application thereof.The mRNA vaccine, characterized in that, contain A27L antigen selected fragment, or L1R antigen selected fragment.The mRNA vaccine provided by the present application successfully induced effective immune response in mice and cattle, detected high level of serum neutralization titer, and the effect is better than goatpox live vaccine (CVCC AV41) vaccine, and the antibody induced has no antibody-dependent enhancement effect.This breakthrough fills the blank in the current LSDV vaccine research and development field, provides a new solution for the production and research and development of animal vaccine, and has very high commercial value and broad application prospect.

The invention relates to the technical field of biology, in particular to a monoclonal antibody for resisting monkey pox virus A29L protein and application of the monoclonal antibody. The monoclonal antibody aiming at the monkey pox virus A29L protein is prepared, and the antibody with high affinity and high specificity is screened by optimizing an antibody screening and expression technology, so that a new tool is provided for rapid detection, effective neutralization and treatment of the monkey pox virus; and an important immunological basis is provided for vaccine research and development of the monkey pox virus and design of a treatment scheme.

The invention belongs to the technical field of quadruple vaccine research and development, and particularly relates to a preparation method of a chicken new flow glandular synovial quadruple vaccine, which comprises the following steps: step one, strain selection: selecting a Newcastle disease LaSota strain, an avian influenza H9 subtype NJ02 strain, a chicken glandular FADV-4 strain and a chicken synovial sac HN01 strain; the selected Newcastle disease strain, the selected avian influenza strain, the selected chicken gland strain and the selected chicken synovial sac strain are inoculated in different culture solutions for culture amplification, the culture environment is controlled to be about 37 DEG C (the temperature before and after the culture is not larger than 0.5 DEG C), and the strain culture time is controlled to be 2-4 days. By selecting the Newcastle disease LaSota strain, the avian influenza H9 subtype NJ02 strain, the chicken gland FADV-4 strain and the chicken synovial sac HN01 strain, the subsequent quadruple vaccine prevention effect can be guaranteed, sufficient quadruple vaccine raw materials can be provided through strain culture, and the production efficiency is guaranteed.

The invention belongs to the technical field of vaccine research and development, and particularly relates to a method for testing the efficacy of a porcine pseudorabies GE gene deletion inactivated vaccine, which comprises the following steps of: inoculating a porcine pseudorabies virus HNQYY2012gE strain virus seed (the virus content per 1ml is more than or equal to 108.5 TCID50) into suspension cultured ST cells according to MOI = 0.05, sampling every 12 hours after virus inoculation, counting and observing the cytopathic effect, and determining the efficacy of the inactivated vaccine with the efficacy of the porcine pseudorabies GE gene deletion inactivated vaccine according to the efficacy of the inactivated vaccine with the efficacy of the porcine pseudorabies GE gene deletion inactivated vaccine and the efficacy of the porcine pseudorabies GE gene deletion inactivated vaccine. The method comprises the following steps of: simultaneously measuring the titer of TCID50, sampling, counting and observing the cytopathy condition 42 hours after virus inoculation, simultaneously immunizing and replacing an animal guinea pig and a target animal pig by using a porcine pseudorabies inactivated vaccine (HNQYY2012gE strain), and researching the correlation between the immune antibody level and the challenge protection of the two animals. A basis is provided for the relevance of an efficacy evaluation method in target animal and alternative animal antibody level and challenge protection, the cost of porcine pseudorabies gE gene deletion inactivated vaccine development can be effectively reduced, and the problem that porcine pseudorabies virus negative pigs are very difficult to screen at present can be solved.

The invention discloses an application of tetrahydrobiopterin in enhancing functions of dendritic cells and relieving related diseases, and finds that tetrahydrobiopterin can promote maturation of the dendritic cells, improve antigen processing and presenting capabilities of the dendritic cells and / or enhance T cell activation capability mediated by the dendritic cells for the first time. The tetrahydrobiopterin can promote synergistic activation of vaccine-induced humoral immunity and cellular immunity by enhancing the dendritic cell function, and the invention provides a new thought and strategy for vaccine research and development, and has important scientific significance and clinical application value.

The present invention provides a dual-modal magnetic particle fluorescence probe and its application in the preparation, tracing and monitoring of dendritic cell vaccines, belonging to the field of biotechnology. The dual-modal magnetic particle fluorescence probe includes: magnetic nanoparticles, cell-penetrating peptides, fluorescence resonance energy transfer fluorescence reporter groups and caspase substrates. The dual-modal magnetic particle fluorescence probe of the present invention combines the advantages of nuclear magnetic resonance imaging and fluorescence resonance energy transfer imaging, can provide real-time imaging information in different biological events, has a high signal-to-background ratio and specificity, and can realize the in vivo monitoring of the behavior of dendritic cell vaccines in real time, including processes such as migration, localization and apoptosis, which helps to improve the efficiency and accuracy of vaccine research and application.

The invention discloses a Zika virus ZIKV (Zika virus, ZIKV) challenging animal model construction method, which comprises the following steps: inoculating ZIKV to an F344 rat through an intracranial injection way for modeling to obtain the ZIKV challenging animal model. After inoculation of ZIKV, obvious ZIKV load can be detected in brain tissues, spleens and reproductive organs of F344 rats, and the load of ZIKV is increased along with prolonging of time. It is shown that the ZIKV challenge model is successfully constructed, and the ZIKV challenge model can be applied to related researches of ZIKV infection mechanisms, vaccine research and development, anti-ZIKV drug research and development and the like.

The invention belongs to the technical field of vaccine adjuvants, and particularly relates to application of tamibarotene and a Toll-like receptor stimulant in preparation of a vaccine adjuvant. The invention relates to a tamibarotene vaccine adjuvant composition, which is prepared by combining tamibarotene and a Toll-like receptor (TLRS) agonist, and the tamibarotene vaccine adjuvant composition is prepared by combining tamibarotene and the TLRs agonist. The composition can significantly promote antigen-specific mucosal immune response, and especially has a prominent effect on gastrointestinal mucosa parts; meanwhile, an organism can be effectively stimulated to generate specific systemic humoral immunity and cellular immune response. In an enterohemorrhagic Escherichia coli challenge model experiment, the vaccine adjuvant composition can be used for remarkably improving the immune protection effect induced by the recombinant intimin antigen, and a new effective strategy is provided for vaccine research and development and immune enhancement.

The invention belongs to the technical field of immunology, and particularly relates to a mouse anti-siniperca chuatsi immunoglobulin M monoclonal antibody and a preparation method and application thereof. The invention provides a monoclonal antibody of anti-immune globulin M. The monoclonal antibody is a monoclonal antibody of anti-immune globulin The monoclonal antibody can accurately recognize natural IgM in siniperca chuatsi serum, does not generate cross reaction with other immune globulins (such as IgT and IgD), and ensures the accuracy of a detection result. Indirect enzyme-linked immunosorbent assay (ELISA) verifies that the antibody shows strong binding activity when recognizing siniperca chuatsi IgM, and is suitable for development of efficient detection kits. The antibody is suitable for various immunological detection methods, such as Western blot, can be specifically combined with a siniperca chuatsi IgM heavy chain, and provides a reliable tool for serum sample analysis. The monoclonal antibody can promote disease prevention and control and vaccine research and development in the aquaculture industry, and has remarkable economic and social benefits.

The invention provides an ELISPOT (Enzyme-Linked Immunospot Assay) analysis method for detecting IFN-gamma and IL-2 secretion, which comprises the following steps: activating an ELISPOT plate, washing, adding IFN-gamma and IL-2 capture antibody coating with specific concentration, and staying overnight at 4 DEG C; recovering and counting target sample cells, resuspending the target sample cells according to a certain concentration, and adding the target sample cells into coated plate holes; incubating for more than 16 hours in an environment of 37 DEG C and 5% COC; washing the incubated plate, adding IFN-HRP and IL-2-ALP detection antibodies, and incubating at room temperature for 2 hours; after washing, adding a blue developing solution for developing for 10-15 minutes; adding a red developing solution, incubating for 10-15 minutes, and stopping developing; according to the method, T cell subset identification and multifunctional analysis can be carried out, in the early stage of vaccine research and development, due to the fact that target positive population is small, the number of obtained PBMC cells is small, two-factor detection can detect different cell factor reaction frequencies and locate different cell populations and subtypes at the same time, and comparison of two-factor data is beneficial to finding out population immune differences; and scientific research and vaccine development are promoted.

The application discloses a recombinant novel goose parvovirus infectious clone plasmid and application, the infectious clone plasmid includes the full-length genome of novel goose parvovirus, and a synonymous mutation marker is introduced at the cysteine codon position in the non-structural protein coding region, under the condition that the amino acid sequence is not changed and the offspring virus replication and infection ability are not influenced, a stable single nucleotide genetic marker is provided for the novel goose parvovirus.The infectious clone plasmid can be used for rescuing the recombinant virus carrying a specific synonymous mutation marker, the obtained recombinant virus has good duck embryo adaptability and cell infection ability, and is suitable for application scenes such as cell infection and immunofluorescence detection.The application provides an important technical tool for novel goose parvovirus genetic marker tracking, pathogenic mechanism research, genetic variation analysis and vaccine research and development.

An adenoviral vector carrying a trichinella spiralis CLP gene, a construction method therefor, and an application thereof, relating to the technical fields of genetic engineering and recombinant viral vaccines. In order to solve the technical problem of existing trichinella spiralis vaccines being administered as a 2 to 3 dose series and failing to provide long-term efficacy, the present invention uses a trichinella spiralis CLP protein as an antigen, and integrates a gene encoding the CLP protein into an adenovirus vector by means of recombinant DNA technology, so as to obtain a recombinant adenoviral vector; the recombinant adenovirus vector is then transfected into mammalian cells and subjected to packaging and amplification processes to obtain a recombinant adenovirus, with multiple experiments then conducted to confirm the recombinant adenovirus. In addition, animal immunization experiments demonstrate that immunizing mice with recombinant adenovirus rAd5TsCLP achieves effective protection against trichinella spiralis infection. The present invention lays a foundation for subsequent trichinella spiralis vaccine research.

The invention discloses a respiratory virus inactivation data-driven modeling prediction control method, which relates to the technical field of biological medicine, and comprises the following steps: acquiring initial virus characteristic data from a virus surface structure database through molecular dynamics simulation, and processing to obtain a structure change matrix of viruses under different inactivation conditions; performing feature extraction on virus characteristics and antibody binding sites by adopting a support vector machine classifier according to the structure change matrix, and determining a potential unexpected interaction site set; according to the respiratory virus inactivation data-driven modeling prediction control method, the scientificity and stability of inactivation process parameter determination are effectively improved, the safety risk caused by improper inactivation conditions is reduced, the process optimization period is shortened, the experiment cost is reduced, the good immune effectiveness is maintained while the virus inactivation safety is guaranteed, and the method is suitable for popularization and application. Therefore, the practical application requirements in vaccine research and development and virus prevention and control can be better met.

The present invention provides a neutralizing monoclonal antibody 10G8 that recognizes CV-A6. The six CDR regions of the monoclonal antibody 10G8 are as follows: (1) the amino sequence of the heavy chain CDR1 is GFTLKNYA; (2) the amino sequence of the heavy chain CDR2 is VSSGGST; (3) the amino sequence of the heavy chain CDR3 is EREEGGYAEAWFAY; (4) the amino sequence of the light chain CDR1 is QSIVQSNGNTY; (5) the amino sequence of the light chain CDR2 is EVS; and (6) the amino sequence of the light chain CDR3 is FQGSHVPFT. The monoclonal antibody 10G8 provided by the present invention can recognize the CV-A6 virus and has neutralizing activity, with a 100% protection rate in mice, and is of great significance for the treatment of diseases caused by the CV-A6 pathogen and vaccine research.

The application provides a recombinant plasmid for expressing HIV P55 protein, a recombinant engineering bacterium and a method for preparing and purifying HIV P55 protein, and belongs to the technical field of protein engineering. The recombinant plasmid takes a PET-30a vector as a backbone plasmid and comprises an expression fragment of the HIV P55 protein. The E. coli Rosetta-PET-30a system adopted in the application has low cost, short culture period (the whole process can be completed within 96 hours), mild induction condition, less degradation of the target protein and can realize large-scale industrial production; the prepared HIV P55 protein has good antigen activity and can be widely applied to the fields of HIV diagnostic kit preparation, subunit vaccine research and development and antiviral drug screening.

The invention relates to the technical field of molecular biology, in particular to an in-vitro transcription method suitable for a wide mRNA length interval, a kit and application. In a transcription reaction system adopted by the in-vitro transcription method, the final reaction concentration of a DNA template is 30-60 [mu] g / mL, the reaction concentration of T7 RNA polymerase is 8-12 U / [mu] L, the reaction concentration of Mg < 2 + > is 40-60 mM, and the reaction concentration of a cap analogue is 3.5-4.5 mM. The method can be applied to a wide mRNA length interval, the application range of mRNA in-vitro transcription synthesis is expanded, the synthesis requirements of mRNA with different lengths are met, the flexibility and universality of mRNA in-vitro transcription synthesis are improved, and a more flexible and universal mRNA synthesis method is provided for the fields of gene therapy, vaccine research and development and the like.