151 results about "High titer" patented technology

The invention relates to the technical field of molecular biology, in particular to an avian adenovirus serum type 4 colloidal gold test strip based on a monoclonal antibody and application of the test strip. According to the invention, two monoclonal antibodies (6B3 and 8G11) aiming at FAdV4 penton protein are successfully prepared through a hybridoma technology, and both the monoclonal antibodies show high titer and strong serotype specificity. Specific analysis proves that the two monoclonal antibodies can recognize 12 FAdV4 isolates from different sources, and do not have cross reaction with other FAdV serotypes or common poultry pathogens. According to the invention, through systematic optimization of key parameters, the detection sensitivity of the test strip is 7.81 * 10 < 4 > TCID50 / 0.1 mL, and the detection time is only 15 minutes. Clinical sample verification shows that compared with an ELISA method, the total coincidence rate is 98.6%.

The invention belongs to the technical field of biochemical engineering, and particularly relates to an amitriptyline hapten and a preparation method thereof, an amitriptyline antigen and a preparation method thereof, an antibody, a kit and application. The amitriptyline hapten has a structural formula shown as a formula (I) or a formula (II). Compared with an existing structure, the amitriptyline hapten prepared by the invention has a better immune effect. The obtained antibody is high in titer, strong in specificity and high in affinity, the LOD of amitriptyline by using an Elisa method established by the antibody is 0.013 ng / mL, the IC50 of amitriptyline is 0.865 ng / mL, the quantitative detection range is 0.07-470.3 ng / mL, the detection sensitivity is high, and the linear range is wide.

The invention discloses chicken infectious anemia virus-like particles as well as a preparation method and application thereof. The invention discloses a virus-like particle vaccine for preventing chicken infectious anemia. The virus-like particle vaccine comprises VP1 and VP2 proteins of chicken infectious anemia viruses. Chicken infectious anemia VP1 and VP2 proteins are expressed by using a baculovirus expression system, immunoblotting shows that the two proteins are successfully expressed in sf9 cells, electron microscope observation finds that the expressed proteins can be autonomously assembled into complete chicken infectious anemia virus-like particles, and the chicken infectious anemia virus-like particles have a space structure similar to that of an original virus and can be used for preparing chicken infectious anemia virus-like particles. Meanwhile, the virus-like particles have the advantages of high titer, high safety, capability of stimulating humoral immunity and cellular immunity and the like. The preparation method disclosed by the invention is simple, can be used for preparing the antigen protein of the chicken infectious anemia virus on a large scale, is high in expression quantity and short in time consumption, greatly reduces the production cost, and is suitable for large-scale production.

The invention discloses an allergen specific IgE and IgG antibody composite quality control product or calibration product as well as a preparation method and application thereof, and belongs to the technical field of biological detection. The preparation method comprises the following steps: obtaining a whole-genome sequence comprising a human IgE constant region gene sequence, a human IgG4 constant region gene sequence and a human IgG1 constant region gene sequence, and preparing a fusion protein simultaneously containing a human IgE Fc segment and a human IgG Fc segment; immunizing a healthy animal by using the allergen to obtain a purified antibody; and carrying out chemical coupling on the purified antibody and the fusion protein to obtain allergen specific IgE and IgG antibodies. By constructing allergen specific IgE and IgG antibodies, the kit can be simultaneously applied to detection of allergens IgE and IgG, has the advantages of high titer, high purity, good dilution linearity and the like, and is beneficial to improvement of diagnosis accuracy and treatment effectiveness.

This invention discloses a hybridoma cell line that secretes anti-polystyrene monoclonal antibodies and its applications. The hybridoma cell line includes PS-8, classified as Hybridoma cell line PS-8. The accession number for PS-8 is CCTCC NO: C2024144, the accession date is May 10, 2024, and the depositary institution is the China Center for Type Culture Collection (CCTCC), located at Wuhan University, No. 299 Bayi Road, Wuchang District, Wuhan City, Hubei Province. The hybridoma cell line provided by this invention can stably secrete anti-polystyrene monoclonal antibodies with high titer, good specificity, and high sensitivity, making it suitable for detection in reagent kits and possessing potential application value.

This invention discloses a broad-spectrum Staphylococcus aureus bacteriophage, SapYZUbeta, and its applications. This bacteriophage has been deposited at the China Center for Type Culture Collection (CCTCC) on May 16, 2022, with accession number CCTCC NO: M 2022629. Identification revealed that the full-length gene of this bacteriophage is 17,790 bp, and it does not contain tRNAs or genes associated with lysogenicity, virulence, or antibiotic resistance. This genetic background confirms the safety of this bacteriophage for use against foodborne pathogens and for bacteriophage therapy. Furthermore, the bacteriophage exhibits a high titer of 1.9 × 10⁻⁶. 9 The phage, with a concentration of PFU / mL, exhibits stable activity within a pH range of 3–12 and a temperature range of -80–60°C, and can be applied to various matrices. The phage provided by this invention can lyse multidrug-resistant wild-type strains isolated from different regions, including MRSA and MSSA, and is characterized by a broad lysis spectrum and high lysis efficiency.

The invention discloses a Nipah virus F protein monoclonal antibody and application thereof, the antibody comprises an antibody 2D12 and an antibody 3B10, and the Nipah virus F protein monoclonal antibody is prepared from Nipah virus F protein immunogen. The specific anti-Nipah virus F protein monoclonal antibody pair is prepared and detected, the monoclonal antibody pair can be specifically combined with the Nipah virus F protein, is high in titer and good in affinity and has neutralizing activity, and meanwhile, the detection kit prepared on the basis of the antibody pair can effectively carry out Nipah virus detection; and technical support can be provided for treatment, epidemiological monitoring and diagnosis of Nipah virus infection.

The application discloses a novel trimeric recombinant protein vaccine. Specifically disclosed are a mutant protein RBD8M with an amino acid sequence of SEQ ID No. 1 and a fusion protein of positions 26-277, positions 1-277 and SEQ ID No. 4. The application also discloses a vaccine containing the fusion protein, and further develops a trimeric recombinant protein vaccine containing the fusion protein and a double adjuvant. Experiments show that the vaccine prepared in the application can effectively induce cellular immunity and humoral immunity of the body in animals, a trace amount of the protein can stimulate a very high immune response of mice, has a good protection effect, can simultaneously produce high-titer neutralizing antibodies against various epidemic strains of the novel coronavirus, is a broad-spectrum effective new coronavirus vaccine, and has important significance for preventing novel coronavirus infection and a wide clinical application prospect.

The present application belongs to the technical field of bioengineering, and particularly relates to a Claudin18.2 gene engineering antibody based on DNA immunization and a preparation method and application thereof. The Claudin18.2 gene engineering antibody comprises a heavy chain variable region and a light chain variable region; the heavy chain variable region comprises any one of the amino acid sequences shown in SEQ ID NO. 15-16; and the light chain variable region comprises the amino acid sequences shown in SEQ ID NO. 19-20. The Claudin18.2 gene engineering antibody is obtained by immunizing Balb / c mice with full-length plasmid DNA and full-length plasmid DNA+extracellular segment polypeptide as immunogens respectively, obtaining plasma cells capable of secreting specific antibodies against Claudin18.2, and then preparing specific mouse-derived gene engineering antibodies through single-cell optical guidance technology and genetic engineering technology. The gene engineering antibodies have high titer and strong specificity, and have specific recognition ability for natural protein Claudin18.2.

This invention relates to the field of bioengineering technology, specifically to a recombinant adenovirus displaying a large antigen fragment on its capsid surface, its construction method, and its applications. The HVR1 and HVR5 regions of the hexon hypervariable region of this virus are replaced by gene fragments containing exogenous genes; each exogenous gene fragment has a flexible linker attached to both ends, and the gene sequence of the flexible linker is shown in SEQ ID No. 1. By precisely locating the naturally variable HVR1 and HVR5 loop regions exposed on the surface of the hexon protein as insertion sites, and supplementing them with flexible linkers, the core problem of exogenous sequence insertion disrupting the stability of the hexon structure is successfully solved. This allows for stable packaging to produce viral particles with high titers and infectivity, enabling the development of a new generation of highly efficient and safe capsid-displaying adenovirus vaccines.

The invention relates to a theophylline immunodetection reagent as well as a preparation method and a detection method thereof, in particular to a theophylline chemiluminescence immunodetection reagent as well as a preparation method and a detection method thereof, and the theophylline chemiluminescence immunodetection reagent comprises an anti-theophylline specific antibody and an indicating reagent for detecting an anti-theophylline specific antibody-theophylline conjugate, the anti-theophylline specific antibody is obtained by immunizing animals with theophylline immunogen. The theophylline immunogen has the beneficial effects that the theophylline immunogen has strong specificity and high immunogenicity, and the prepared anti-theophylline specific antibody has strong specificity and high titer; the chemiluminescence immunoassay reagent containing the anti-theophylline specific antibody can conveniently, rapidly and accurately determine the theophylline content in a sample, can simultaneously determine a plurality of samples on a full-automatic chemiluminescence immunoassay instrument, realizes high-flux rapid determination of theophylline, and has the advantages of high accuracy, strong specificity, high sensitivity and high sensitivity. And the accuracy and the detection efficiency are greatly improved.

The application discloses a kind of immortalization human umbilical cord mesenchymal stem cell NOTCH1-LNR overexpression stable cell line construction method and its application in liver differentiation, it is related to immortalized cell technical field.The application first connects target gene to vector, and the slow virus plasmid containing target gene is constructed, the slow virus plasmid containing target gene is introduced into 293T cell, and high titer slow virus containing target gene is generated;Immortalized human umbilical cord mesenchymal stem cell is infected with slow virus, and mixed clone cell strain is screened, and the immortalized human umbilical cord mesenchymal stem cell NOTCH1-LNR overexpression stable cell line is obtained.The application proves that the differentiation efficiency of immortalized human umbilical cord mesenchymal stem cell NOTCH1-LNR overexpression can effectively improve liver-like cell to glycogen staining and Western blotting experiment.This research result is expected to provide new technical thought for the next step development to improve the yield of MSCs liver differentiation.

Provided are a set of fully human TSH receptor (TSHR)-blocking monoclonal antibodies targeting a CHK36 homolog cluster, a preparation method therefor, and an application thereof. The method comprises the following steps: using flow cytometry to sort plasma cells and single memory B cells that specifically recognize TSHR in peripheral blood of a patient with a high titer of TSH-blocking antibodies (TBAb), performing in vitro cloning of antibody light and heavy chains and performing recombinant expression, and using hTSHR-CHO cells to perform screening and validation of antibody properties to obtain a blocking monoclonal antibody that specifically targets human TSHR. The fully human TSH receptor-blocking monoclonal antibodies specifically bind to a TSHR, and effectively block signal transduction after a TSH binds to a receptor, inhibit the synthetic secretion of thyroid hormones, and significantly reduce and inhibit TSHR expression and fibrosis in orbital fibroblasts of effector cells associated with thyroid-related ocular diseases. The fully human TSH receptor-blocking monoclonal antibodies have broad application prospects in the treatment of Graves' disease (GD) and other diseases caused by hyperthyroidism, such as thyroid eye disease (TED).

The application belongs to the technical field of bioengineering, and particularly relates to a bacteriophage PE-2 of Escherichia coli with high blood entry efficiency and fermentation efficiency and application. The bacteriophage involved in the application has a preservation number of CCTCC NO: M20252973, and can specifically lyse pathogenic Escherichia coli O2 serotype and has high blood entry efficiency. The bacteriophage can enter blood circulation through oral administration, and can be widely distributed to the spleen, liver, heart and other tissues through blood circulation. The bacteriophage can be prepared in pilot production to obtain a very high titer, and the titer reaches 1.6x10 12 PFU / mL. Compared with the production of antibiotics, the production cost of the bacteriophage is low, and the bacteriophage provides a source and a reference example for industrial production of high-titer bacteriophage.

The application discloses an immunogen for preparing testis-specific glyceraldehyde-3-phosphate dehydrogenase lysine malonylation site-specific antibody, and belongs to the technical field of biotechnology.The immunogen is a malonylation modified polypeptide, and the amino acid sequence is IIPASTGAAKAVT, corresponding to the 278th-290th amino acid of a human GAPDHS protein, wherein a lysine at the 287th position (the 10th position in the sequence) is subjected to malonylation modification.A specific antibody designed according to the immunogen has the characteristics of high titer and high specificity. By screening a human sperm sample by using the antibody, it is found that the antibody can not only specifically recognize the GAPDHS with the lysine at the 287th position subjected to malonylation modification in vitro and in human sperm, but also is closely related to sperm forward movement and asthenozoospermia. The application has great significance in the field of preparation of reagents for precision diagnosis and treatment of idiopathic male infertility.

The inventions provide improved systems and methods for inline concentrating and reducing intermediate process pools to mitigate plant fit challenges associated with high-titer protein purification process. The inventions also provide methods for concentrating and reducing VI pool volumes by integrating columns of single pass tangential flow filtration (SPTFF) system with viral inactivation pool filtration (VIPF) in protein purification process. Therapeutic proteins purified by the methods also are provided.

The invention discloses preparation and application of a mouse TMC5 truncated recombinant protein and a polyclonal antibody, and belongs to the technical field of biological medicine. The amino acid sequence of the TMC5 truncated recombinant protein is as shown in SEQ ID NO. 1, and the nucleotide sequence of the TMC5 truncated recombinant protein is as shown in SEQ ID NO. 2. Compared with a full-length protein, the mouse TMC5 truncated recombinant protein provided by the invention has better stability and antigenicity, the obtained polyclonal antibody is high in titer and strong in specificity, the identification of endogenous TMC5 protein in mouse testicular tissues is facilitated, meanwhile, the TMC5 truncated recombinant protein and the polyclonal antibody can also be used for researching a mechanism of regulating spermatogenesis by TMC5, and the application prospect is broad. The blank of preparation of the mouse TMC5 antibody and functional research of the mouse TMC5 antibody in the testis is filled. The mouse TMC5 truncated recombinant protein and the polyclonal antibody can be applied to development of novel diagnostic reagents and drugs for treating male sterility.

The invention belongs to the technical field of veterinary biology, and particularly relates to a swine foot and mouth disease virus type A immune peptide fusion protein as well as a preparation method and application thereof. The invention provides a swine foot-and-mouth disease virus type A immune peptide fusion protein. The swine foot-and-mouth disease virus type A immune peptide fusion protein has an amino acid sequence as shown in SEQ ID NO: 1. Compared with an epitope predicted in the prior art, the recombinant immune composite peptide provided by the invention has better immunogenicity, can obtain a neutralizing antibody with higher titer, and has better animal protection capacity.

The invention relates to a polyclonal antibody of enterohemorrhagic Escherichia coli Intimin protein as well as a preparation method and application of the polyclonal antibody, and the polyclonal antibody of the enterohemorrhagic Escherichia coli Intimin protein can specifically recognize the enterohemorrhagic Escherichia coli Intimin protein and has relatively high titer; according to the present invention, with the enterohemorrhagic Escherichia coli Intimin protein polyclonal antibody, the infection process of enterohemorrhagic Escherichia coli can be deeply understood through the enterohemorrhagic Escherichia coli Intimin protein polyclonal antibody, such that the hereditary basis and the molecular mechanism of the enterohemorrhagic Escherichia coli infection host cell can be further researched; the method has double values for understanding pathogenesis and developing prevention and control means.

The present application relates to the technical field of central obesity related research and treatment, by constructing a central obesity mouse model, it is found that the up-regulation of brain-derived neurotrophic factor (BDNF) expression in adipose tissue is closely related to central obesity resistance. A BDNF fusion polypeptide composed of TAT cell-penetrating peptide, flexible linker and BDNF mature peptide is designed and prepared, the fusion polypeptide has strong cell viability promoting effect at the cellular level, can effectively activate the BDNF signaling pathway and has better stability than natural BDNF protein; an anti-TrkB monoclonal antibody with high affinity and high titer is also prepared. In vivo experiments show that the BDNF fusion polypeptide and its combination with the anti-TrkB monoclonal antibody can effectively inhibit the weight gain of central obesity mice, improve glucose metabolism disorder, increase serum BDNF level and activate the TrkB signaling pathway, and the combination has better synergistic therapeutic effect, which provides a new direction and experimental basis for the treatment of central obesity.

This invention relates to the field of biomedical technology, providing a rabbit hemorrhagic disease virus (RHDV) type 2 fusion antigen protein and its applications. The fusion antigen protein comprises the RHDV2 VP60 protein sequence and an immunomodulatory CRM197 protein functional fragment, fused together via a linker peptide, and expressed and purified in an insect cell expression system. This invention improves the immunogenicity of the antigen protein by fusing the major capsid protein VP60 of RHDV2 with the immunomodulatory fragment of CRM197. Rabbit immunization experiments verified that the fusion antigen protein can induce high-titer specific antibodies, with serum antibody titers reaching 1:12800 and above, demonstrating good immunization effects. The fusion antigen protein of this invention can be used to prepare a vaccine for the prevention of RHDV2 infection, providing a new technical solution for the prevention and control of RHDV2 disease.

The invention discloses a non-human animal integrated with a double-derived VH subclass fusion gene as well as a construction method and application of the non-human animal. The method comprises the following steps: firstly, constructing a gene construct containing human IGHV1-69 and IGHV3-23 fragments, a specific spacer region, a functional element and a rat heavy chain 3'enhancer, and microinjecting the gene construct into a C57BL / 6J fertilized egg with an endogenous heavy chain and a kappa light chain knocked out to obtain a transgenic mouse; through identification, after a mouse endogenous antibody is inactivated, a human gene is stably integrated, a human IgM antibody is expressed and H5N1 HA immunization is carried out, the use frequency of IGHV1-69 is remarkably increased, the mouse produces a high-titer specific antibody, and serum has neutralizing activity. The mouse can generate a target bnAbs aiming at the HA stem and the RBS in parallel, so that the antibody screening efficiency is improved, and an in-vivo screening and evaluation platform for simulating human body fluid response is provided for research and development of influenza antibodies and vaccines.

The present invention relates to the biomedical and biopharmaceutical sectors. Specifically, it relates to a chimeric antigen comprising the combination of the amino- and carboxy-terminal regions of the amyloid-beta peptide (Aβ), the amino- and carboxy-terminal regions of the tau protein, and a T-cell epitope. The pharmaceutical composition comprising this chimeric antigen and at least one pharmaceutically acceptable vaccine adjuvant increases the efficacy of immunotherapy for the prevention and treatment of Alzheimer's disease (AD). The chimeric antigen exerts its action by stimulating a multi-target humoral response with high titers of anti-Aβ and anti-tau antibodies simultaneously. This promotes the combined elimination of toxic Aβ and tau species from the brain, thereby preventing or significantly improving the clinical symptoms and neuropathology of AD.

The present invention describes an efficient vaccine against a Chlamydia trachomatis (Ct). The vaccine is based on recombinant fusion molecules that are capable of generating a high titered neutralizing antibody response that is protective against various Ct serovars. Our invention furthermore describe the combination of these antibody promoting fragments with Ct antigens that are targets for T cells with the aim to provide a vaccine that activate both arms of the immune system.

The application belongs to the technical field of genetic engineering and microbial metabolic engineering, and particularly relates to a modular construction method of a high-titer LanM protein engineering strain. The construction method realizes high-level soluble expression of LanM protein in Escherichia coli through four-dimensional modular synergistic regulation of spatial positioning optimization, chaperone-assisted folding, central carbon metabolic flow redistribution and amino acid synthesis strengthening. The system not only improves the yield of the target protein, but also provides a technical reference for the heterologous expression of other metal-binding proteins with similar structural characteristics. Through the synergy of plasmid copy number control, promoter strength matching and induction timing between the modules, the dynamic balance of intracellular metabolic flow and protein synthesis flow is achieved, meeting the requirements of industrial large-scale production.

The invention provides a monoclonal antibody of NMN and application and a product thereof, and relates to the technical field of biology. According to the NMN monoclonal antibody provided by the invention, the research of the inventor finds that the monoclonal antibody can specifically recognize NMN, the properties of high titer, good gradient, specificity, repeatability, stability and the like of the monoclonal antibody meet the requirements of an NMN detection kit, and the monoclonal antibody can be used for diagnosing pheochromocytoma.

The application provides an anti-glycocholic acid antibody and application thereof, and specifically, the application immunizes BALB / C mice with a glycocholic acid antigen, and more than ten positive monoclonal hybridoma cell strains are screened and obtained. Through performance determination, finally, 3 cell strains capable of specifically combining with glycocholic acid are obtained. Among them, the H15 antibody has high titer, good stability, and stable detection linearity, and can be used for clinical detection of glycocholic acid.

The invention is applicable to the field of genetic engineering and biotechnology detection, and provides a nano antibody specifically bound with Seneca virus A and application of the nano antibody. The nucleotide sequence of the nano antibody is as shown in SEQ ID NO: 1, and the amino acid sequence of the nano antibody is as shown in SEQ ID NO: 2. The anti-SVA nano antibody provided by the invention is relatively high in titer and good in specificity; the detection method provided by the invention is used for detecting the Senecavirus A antibody, and has good specificity and sensitivity.

The invention discloses a pseudomonas syringae phage as well as a composition and application thereof, and belongs to the technical field of biological prevention and control. The pseudomonas syringae bacteriophage PhiPs-1 which is short in incubation period, short in outbreak period, large in splitting quantity, high in titer, wide in host spectrum, acid-resistant, heat-resistant, chloroform-resistant and ultraviolet-resistant is obtained through screening, and the preservation number of the pseudomonas syringae bacteriophage PhiPs-1 is CCTCC NO: M 20252347; the composition can be developed into a medicinal preparation for preventing and treating the tomato bacterial leaf spot disease; according to the invention, the pseudomonas syringae phage phi Ps-1 and the biocontrol bacterium NBL-AP73 are combined for use, so that the control effect on the tomato bacterial leaf spot disease is synergistically improved.

The present application belongs to the field of biomedical technology, and more particularly relates to the application of Periplaneta americana polypeptide HFDT1 as an immune adjuvant in the preparation of a brucellosis vaccine. The present application uses Brucella OMP10, BP26 and L7 / L12 proteins in combination with HFDT1 to induce mice to produce higher titers of anti-OMP10, BP26 and L7 / L12 protein specific IgG antibodies, and the titers increase with the increase in the number of immunizations, while promoting the body to produce mixed Th1 / Th2 type cellular immune responses and stimulating the proliferation and differentiation of mouse spleen T and B cells, and also facilitating mass production. The Periplaneta americana polypeptide HFDT1 described in the present application is obtained by convenient mass synthesis, and can be used as a novel small molecule immune adjuvant to enhance specific immune responses of the body.