38 results about "Antiserum" patented technology

The application belongs to the technical field of biotechnology, and particularly relates to a human metapneumovirus mouse-adapted strain and application thereof. Compared with wild strains, the strain can exhibit stronger susceptibility to animals, and can cause typical clinical symptoms and even death. For example, after the strain infects BALB / c mice, the disease symptoms are obvious, 83% of the mice have a severe decrease in body weight to below 75% after infection, and have reached the humane killing standard. Four days after the strain infects BALB / c mice, the mortality rate is above 50%. The strain can be used to construct a cell model or an animal model of human metapneumovirus infection or a disease caused by the human metapneumovirus, and is further used to screen drugs for preventing and / or treating human metapneumovirus infection or a disease caused by the human metapneumovirus. In addition, the strain can also be used to prepare drugs for preventing and / or treating human metapneumovirus infection or a disease caused by the human metapneumovirus, and to prepare antibodies or antisera against the human metapneumovirus.

The invention relates to the technical field of biological medicines, and discloses a construction method and expression application of an Hcp gene deleted strain of a rahnella aqua KCL-5 strain for fish. The Hcp 0186 gene deletion strain is constructed by using a homologous recombination method, the comparison between the deletion strain and a wild strain is carried out, and the result shows that the deletion of the Hcp 0186 gene has an influence on the growth speed, the pathogenicity of the Hcp 0186 gene to the zebra fish is obviously reduced, the survival rate of the infected zebra fish is increased from 0% to 52%, and the Hcp 0186 gene deletion strain can be used as a fish attenuated live vaccine. A recombinant prokaryotic expression vector pET32a-Hcp and expression and purification of fusion protein of the recombinant prokaryotic expression vector pET32a-Hcp are further constructed, and Hcp 0186 rabbit polyclonal antibody serum is prepared as a primary antibody and can be applied to localization expression analysis of fish tissues infected by the bacterium and detection of WB and IHC. Results prove that the Hcp 0186 gene deletion of the strain provides an attenuated strain vaccine and clinical diagnosis and detection application of infected fish of the attenuated strain vaccine, and a scientific basis can be provided for revealing a molecular pathogenic mechanism of the strain and development and application of a mutant strain of the strain.

The invention discloses a crassostrea hongkongensis NKA-alpha protein polyclonal antibody as well as a preparation method and application thereof. The preparation method comprises the following steps: constructing a recombinant expression vector containing an NKA-alpha target gene segment; transforming the recombinant expression vector into competent cells to construct a recombinant expression strain, inducing the recombinant expression strain to express fusion protein, and collecting and purifying to obtain NKA-alpha recombinant protein; carrying out animal immune treatment by taking the NKA-alpha recombinant protein as an antigen, collecting and separating antiserum, and purifying to obtain the Crassostrea hongkongensis NKA-alpha protein polyclonal antibody. The prepared Crassostrea hongkongensis NKA-alpha polyclonal antibody is high in affinity, can specifically recognize NKA-alpha protein in Crassostrea hongkongensis, and can be widely applied to Crassostrea hongkongensis biological immunodetection.

The invention relates to key antigen epitope screening, in particular to an echovirus 18 type key antigen epitope peptide and application thereof. The result of an enzyme-linked immunosorbent assay shows that Epitope 1 (SEQ ID NO: 1) can be effectively combined with E18-VP1 antiserum, and the combining capacity is obviously higher than that of Epitope 2 in a blank control and a negative result, so that one antigen epitope peptide is screened based on the complete sequence of the echovirus 18-VP1, and the antigen epitope peptide has important guiding significance for research and development of echovirus 18 vaccines.

This invention relates to a high-titer prealbumin antibody purification method and its application, belonging to the field of biotechnology. The method utilizes Protein A affinity chromatography to capture total IgG from antiserum, followed by purification using an affinity chromatography column prepared by targeted conjugation of a specific sequence peptide. The antibody purified by this method achieves a titer of 1:256,000 in ELISA detection, with a half-maximal effective concentration (EC50). 50 The antibody concentration was 3.2 ng / mL, exhibiting extremely high affinity and reactivity. When used in a double-antibody sandwich chemiluminescent immunoassay, it achieved a detection limit of 0.08 µg / mL, a wide dynamic range (0.08–400 µg / mL), and a titer retention rate as high as 88% after accelerated storage at 37°C for 14 days, demonstrating significantly better stability than antibodies prepared by traditional methods. This invention provides a highly active and stable core antibody raw material for the preparation of high-performance immunodiagnostic reagents.

The application provides a non-pH-dependent long-acting anti-serum albumin nanobody and an application thereof. Specifically, the application provides amino acid sequences of VHH chain framework regions FR and complementarity determining regions CDR of the non-pH-dependent long-acting anti-serum albumin nanobody. The application also provides nucleotide sequences encoding the nanobody. The anti-serum albumin nanobody provided by the application can bind to human, mouse, rat and cynomolgus serum albumin under different pH conditions, and can significantly prolong the half-life of a protein drug, thereby providing a research basis for long-acting protein drug development.

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

The application discloses an anti-serum albumin nanobody 3A12 and long-acting application thereof, and belongs to the technical field of antibody engineering. The nanobody complementarity determining region comprises CDR-H1 shown in SEQ ID NO. 2 (SGYIA), CDR-H2 shown in SEQ ID NO. 3 (GIYRLGSSTFYADSVKG), and CDR-H3 shown in SEQ ID NO. 4 (GSTDRSLLRKPDTFNY). The nanobody is coupled with a therapeutic polypeptide, protein or antibody drug to construct a long-acting complex. The complex can significantly delay the renal clearance rate of the drug, resist protease degradation, increase the half-life of the drug, thereby avoiding frequent administration, maintaining the curative effect while reducing the administration frequency, and improving the patient compliance and treatment safety. The application is particularly suitable for the development of long-acting polypeptide, protein and antibody drugs.

The invention belongs to the technical field of biological medicines, and particularly relates to a human NSUN2 protein 190th lysine site lactic acid antigen, an antibody, a preparation method and application thereof. Relates to the technical field of lactylated antibody preparation, and the preparation method comprises the following steps: S01, carrying out lactylation on 190th lysine in a polypeptide of an amino acid sequence, and coupling with a carrier protein to obtain an initial antigen, the carrier protein comprising hemocyanin KLH; s02, injecting the obtained antigen into a living body culture sample for immune culture, and collecting whole blood after a preset period to obtain multi-antibody serum; and S03, purifying to obtain an iterative antibody. According to the invention, the expression difference in sensitive and drug-resistant tumor cells and tumor tissues can be detected by directly applying Western Blotting, immunofluorescence and immunohistochemistry, so that a powerful guarantee is provided for the detection of tumor sensitivity.

The present invention relates to anti-serum albumin antibodies and multispecific binding proteins comprising the same. The present invention also relates to pharmaceutical compositions comprising the antibodies or multispecific binding proteins, expression vectors and host cells for making the antibodies or multispecific binding proteins, and methods of using the antibodies or multispecific binding proteins for treating diseases or conditions.

The invention discloses a method for preparing a high-titer polyclonal antibody from a Zika virus strain, which comprises the following steps: selecting the Zika virus strain, culturing in cells, and purifying virus particles with high purity to prepare an immunogen; an optimized animal immunization strategy is adopted and comprises the steps of selecting a proper host animal, applying a potent immunologic adjuvant, namely a Freund's adjuvant, and designing a scientific immunization program, so that humoral immune response of the host is stimulated to the maximum extent; when the titer of the antibody reaches a peak value, collecting antiserum, synthesizing hybridoma cells through splenocytes, and obtaining the high-purity polyclonal antibody through a multi-step purification technology. According to the method, high-purity and non-chemically-inactivated whole virus particles which retain natural conformation are used as immunogens, a mode of primary immunization and multiple booster immunization is adopted, a Freund's adjuvant is combined, animals are continuously stimulated to generate high-titer antibodies, non-specific antibodies and impurities are effectively removed through multi-step purification, and the immunogen immunogen is obtained. The target antibody with high purity and high specificity is obtained.

This invention relates to the field of biochemical technology, and particularly to highly specific haptens of polymyxin B and colistin, artificial antigens, their preparation methods, and applications. The haptens have structures as shown in Formula I or Formula II. The applications include: (1) preparing specific antibodies against polymyxin B or colistin; (2) detecting specific antibodies against polymyxin B or colistin; and (3) preparing reagents for detecting specific antibodies against polymyxin B or colistin. This invention further provides artificial antigens obtained by conjugating the haptens with carrier proteins. Using the artificial antigens provided by this invention as immunogens to immunize experimental animals, highly specific and sensitive antiserum can be prepared, which can then be used to extract and prepare polyclonal antibodies. These methods can be used to establish highly specific, highly sensitive, simple, and rapid detection methods for polymyxin B and colistin, and have significant application value.

The application relates to the technical field of medical devices, and discloses a protein analysis device for HbA1C antiserum buffer, which comprises an analysis machine main body and a bar code input mechanism installed in the inner cavity of the analysis machine main body, and further comprises: a mounting frame installed on one side of the inner cavity of the analysis machine main body and corresponding to the position of the bar code input mechanism; an electric telescopic rod installed in the interior of the analysis machine main body and corresponding to the position of the bar code input mechanism; and a dismounting assembly arranged on the output shaft of the electric telescopic rod. The space posture of a test tube is adjusted in real time through the cooperative driving of the electric telescopic rod, a micro motor and a rotating roller, so that the surface label of the test tube and the scanning area of the bar code input mechanism are accurately aligned. The foreign matters on the test tube and the surface label can be effectively removed, and the raised label edge can be physically smoothed. The auxiliary mechanism can be regularly cleaned and disinfected through the linkage mechanism of the electric telescopic rod and the cleaning mechanism.

The present invention relates to a nanobody or antigen-binding fragment thereof that specifically binds to serum albumin, a derivative comprising the nanobody or the antigen-binding fragment thereof, a nucleic acid encoding the antibody or antigen-binding fragment thereof, a host cell comprising same, and related uses. Further, the present invention relates to therapeutic use of the antibody or the antigen-binding fragment thereof or the derivative.

The invention relates to the technical field of biological medicine, in particular to a cupreuloplasmin detection kit and a preparation method thereof, the kit comprises a reagent R1 and a reagent R2; the reagent R1 is prepared from a tris (hydroxymethyl) aminomethane buffer solution, polyethylene glycol-8000, sodium azide, tween-80 and magnesium acetate; the reagent R2 is prepared from a trihydroxymethyl aminomethane buffer solution, tween-20, sodium azide, goat anti-human copper cyanin antiserum, bovine serum albumin and L-proline. According to the invention, the gradient surfactant design and the metal ion auxiliary antigen conformation stability are introduced, so that the detection sensitivity and the anti-interference capability are synergistically improved; by constructing a ternary antibody stabilization system, the long-term stability of the reagent is remarkably enhanced, and the batch-to-batch difference is reduced. The kit has the advantages of high sensitivity, strong specificity, good stability and batch-to-batch consistency.

The invention relates to a multivalent vaccine, in particular to a chimera multivalent vaccine based on Ercovirus type 11, 18 and 30 key epitopes as well as a preparation method and application of the chimera multivalent vaccine. According to the invention, a chimera multivalent vaccine VP1-III is also designed based on key epitopes, an E11-VP1 sequence is taken as a basis, one E18-VP1 key epitope and two E30-VP1 key epitopes are respectively inserted into the E11-VP1 sequence to obtain a chimera multivalent protein vaccine VP1-III, and the chimera multivalent protein vaccine VP1-III is taken as an immunogen to immunize a mouse to detect antibody titer and virus neutralizing capacity of antiserum. A new thought is provided for designing and preparing multivalent vaccines and detecting immunogenicity of viral vaccines.

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.

Provided are modified cells and cell lines containing extracellular enveloped viruses (EEV), methods of producing serum-resistant EEVs, and programmed EEVs for systemic oncolytic viral therapy. The viruses are high EEV producing viruses that contain genome modifications that alter properties and / or activities of the viruses. The cells and cell lines are modified to encode polypeptides or peptides that are displayed on their membranes, which, when virus is propagated in the cells, encases the virus so that the polypeptides and peptides from the cell membrane are displayed on the virus. The viruses, thus contain modified outer envelopes that display polypeptides and / or peptides encoded by the virus and from the cell membranes. This combination provides viruses whose properties and activities can be programmed by virtue of the cells in which they are produced and viral genome modifications. The viruses can be used for treating cancers or, because the properties of the virus, including tropisms, other diseases, disorders, and conditions involving targeted cells.

The application provides a non-pH-dependent long-acting anti-serum albumin nanobody and application thereof. Specifically, the application provides amino acid sequences of VHH chain framework regions FR and complementarity determining regions CDR of the non-pH-dependent long-acting anti-serum albumin nanobody. The application also provides nucleotide sequences encoding the nanobody. The anti-serum albumin nanobody provided by the application can bind to human, mouse, rat and cynomolgus serum albumin under different pH conditions, and can significantly prolong the half-life of a protein drug, thereby providing a research basis for long-acting protein drug development.

A novel vaccine and methods of preventing and treating C. difficile infection in a patient is described. The vaccine is comprised of a fusion protein (denoted FliCD) comprised of the FliC and FliD from C. difficile and joined by a linker sequence. Administration of the vaccine, as well as anti-FliCD serum, has been shown to prevent C. difficile infection as well as treat existing infections.

The invention relates to the technical field of molecular biology and immunology, in particular to a bovine herpesvirus type 4 gb protein recombinant expression method and application thereof.The 104-384aa segment (the amino acid sequence is shown as SEQ ID NO.2) of bovine herpesvirus type 4 gb protein is selected and constructed to a pet22b / pet32a escherichia coli expression vector, after induced culture and ultrasonication, Ni-NTA gradient elution purification is conducted, and the recombinant expression vector is obtained. The recombinant gb protein with the molecular weight of about 48.5 kDa is obtained. The recombinant protein is used for preparing mouse antiserum, and the titer of the antiserum is not lower than 1: 128000 through ELISA (enzyme-linked immuno sorbent assay) detection; the purity of the purified antiserum meets experimental requirements, and the antiserum can specifically recognize a target antigen. The method solves the problems of poor protein dissolution and low antiserum titer in the traditional method, is controllable in process and high in cost effectiveness, and provides a reliable tool for molecular detection and immunoassay of the bovine herpesvirus type 4.

Anti-Trop2 antibodies, conjugates comprising the antibodies, and uses thereof are provided. Specifically, the invention provides an anti-Trop2 single-domain antibody, a long-acting anti-Trop2 antibody formed by connecting the anti-Trop2 single-domain antibody and an antiserum albumin single-domain antibody in series, and an antibody-drug conjugate constructed based on the long-acting anti-Trop2 antibody, and also provides a coding sequence for coding the antibody, a corresponding expression vector and a host cell capable of expressing the antibody. The anti-Trop2 antibodies and conjugates thereof can be used for the treatment or diagnosis of Trop2 related diseases.

The application discloses an anti-serum albumin nanobody 1B7 and application thereof in prolonging the half-life of a polypeptide drug, and belongs to the technical field of antibody engineering. The nanobody has the following complementarity determining regions (CDRs): CDR-H1 shown in SEQ ID NO. 2 (YTCMG), CDR-H2 shown in SEQ ID NO. 3 (TIDNSGGRTYYADSVKG), and CDR-H3 shown in SEQ ID NO. 4 (QSSGYCLSDNFFRH). The nanobody is coupled with a therapeutic polypeptide, protein or antibody drug by gene fusion technology to construct a long-acting complex. The complex can significantly delay the kidney clearance rate of the drug, resist protease degradation, prolong the half-life of the drug, avoid frequent administration, maintain the curative effect while reducing the administration frequency, and improve the patient compliance and treatment safety.

The invention relates to the technical field of biology, in particular to a preparation method of a high-titer immunocompetence SlgA polyclonal antibody. The invention discloses a preparation method of a high-titer immunocompetence SlgA polyclonal antibody, which comprises the following steps: (1) emulsifying a bovine colostrum SIgA antigen with the purity of more than 95% to obtain an antigen emulsion; (2) injecting an immune animal with the antigen emulsion for multiple times to obtain SIgA antiserum; (3) purifying the SIgA antiserum to obtain an SIgA polyclonal antibody; the SIgA antigen is extracted from bovine coloctrum, the high-titer rabbit anti-bovine SIgA polyclonal antibody is successfully prepared through the method, the antigen immunization dosage is reduced, meanwhile, the high-titer antibody can be obtained, the immune double diffusion titer of the antibody is 1: 16, the ELISA titer is larger than 512000, and the method is a new effective antibody preparation way.

The application relates to the technical field of serum separation, and discloses a separation device and a separation method for preparing a CCP antiserum buffer, which comprises a separator body, a cover plate is rotationally arranged on the surface of the separator body, a rotating machine is electrically connected to the inner cavity of the separator body, a lifting mechanism is arranged on the output shaft of the rotating machine, and a plurality of sleeves for placing serum test tubes are arranged on the surface of the lifting mechanism; when the application is used, the technical pain point that the existing device is inconvenient to take out the test tubes is solved; through the multi-stage lifting mechanism, the sleeves drive the test tubes to rise after centrifugation is completed, the operation space for taking out the test tubes is significantly expanded, the operator can avoid reaching into a narrow cavity to take out the test tubes, the rib on the surface of the handle is matched, the holding friction is increased, the test tubes are effectively prevented from slipping and toppling due to condensed water, the loss and pollution of serum samples are reduced, the convenience and safety of taking out the test tubes are improved, and the taking-out requirement of the CCP antiserum batch preparation is met.

The invention relates to a blood typing kit designed to make blood typing more flexible, whether in or out of the laboratory. The kit comprises coverslips containing ready-made antibodies of anti-A, anti-B, and anti-D sera, a lancet in addition to the coverslips, and compressed alcohol cotton for sterilization.

The invention provides a preparation method of an ictalurus punctatus IgM antibody based on a prokaryotic expression system, and relates to the field of aquatic animal immunodetection and antibody preparation, and the preparation method comprises the following steps: designing a specific primer based on an ictalurus punctatus IgM protein gene sequence in GenBank, and carrying out antigen determinant truncation; the method comprises the following steps: extracting liver RNA (Ribonucleic Acid) of channel catfish by a Trizol method, reversely transcribing the liver RNA into cDNA (Complementary Deoxyribonucleic Acid), and cloning an IgM The method comprises the following steps: constructing a pET-30a-IgM recombinant expression vector, transforming Escherichia coli Rosetta competent cells, and inducing protein expression; purifying the truncated IgM protein by adopting the steps of ultrasonication, affinity chromatography and dialysis concentration; a New Zealand white rabbit is immunized to prepare antiserum, and the titer is detected through ELISA. According to the method, efficient and low-cost preparation of the IgM protein is realized by utilizing a prokaryotic expression system, the period is only several weeks, high-purity protein of milligram-grade to gram-grade can be obtained, the batch stability is good, related biological safety risks of animal serum are avoided, and a key basis is provided for development of an ictalurus punctatus ELISA (enzyme-linked immunosorbent assay) kit, immunohistochemistry and B lymphocyte research.

The application discloses a duck BRS3 polypeptide and a preparation method and application of a polyclonal antibody thereof, and belongs to the field of biochemistry and molecular immunology. The amino acid sequence of the duck BRS3 polypeptide is shown as SEQ ID NO. 3. A polypeptide sequence with a length of 20 aa is screened from an intracellular region of a duck BRS3 protein sequence, and after being artificially synthesized by a solid phase, the polypeptide sequence is used as an immunizing antigen to immunize New Zealand white rabbits, and a duck BRS3 protein polyclonal antibody with an antiserum titer of 1:25600 or above is obtained. The rabbit anti-duck BRS3 polyclonal antibody prepared in the application can specifically combine with the BRS3 protein in duck tissues, fills a blank in the field of duck BRS3 protein detection research, and lays a foundation for the research on the function of BRS3 in a duck body.

This invention discloses a method for preparing and applying a duck interneuron B modified polypeptide and its polyclonal antibody, belonging to the fields of biochemistry and molecular immunology. The amino acid sequence of the duck interneuron B modified polypeptide is shown in SEQ ID NO.4. This invention screens a 19aa polypeptide sequence from the duck interneuron B protein sequence, adds a cysteine ​​residue (Cys, C) to its C-terminus, synthesizes it artificially in a solid phase, and uses it as an immunogenic antigen to immunize New Zealand white rabbits, obtaining a duck interneuron B polyclonal antibody with an antiserum titer of over 1:204800. The rabbit anti-duck interneuron B polyclonal antibody prepared by this invention can specifically bind to interneuron B protein in duck tissues, filling a gap in the research field of duck interneuron B protein detection and laying the foundation for the study of the function of interneuron B in ducks.

The invention relates to the technical field of biology, in particular to a combined culture medium of chicken embryo primordial germ cells (Primordial Germ Cells, PGCs) and a cell culture method of the combined culture medium, calcium ion-free DMEM is used as a basis, various supplements, nucleoside, sodium pyruvate, amino acid, vitamins, additives, trace components, three antibodies, serum and specific cell factors are added, filtration sterilization is performed after osmotic pressure is adjusted, and the combined culture medium and the cell culture method of the combined culture medium are used for culturing the chicken embryo primordial germ cells (Primordial Germ Cells). The cell factors are added before use to adapt to the culture of the related PGCs. The method does not need a feeding layer, simplifies the operation, avoids exogenous pollution, improves the purity and survival state of the PGCs, promotes proliferation, has no sex preference, is adaptive to different sources of PGCs, is stable and reliable, and has a wide application prospect.