75 results about "Secretory antibody" patented technology

The invention relates to murine / human chimeric monoclonal antibodies with high specificity to and affinity for human carcinoembryonic antigen (CEA), derivatives thereof, processes for the preparation of these antibodies and their derivatives, DNAs coding for heavy and light chains of these antibodies, processes for the preparation of said DNAs, mammalian cell lines that produce and secrete the antibodies and processes for the preparation of said cell lines. The chimeric antibodies and their derivatives are used for clinical purposes in vitro and in vivo, especially for the diagnosis of cancer, for localization and in vivo imaging of tumors, for therapy, e.g. site-directed delivery of cytotoxins, and similar purposes. The invention also concerns test kits and pharmaceutical compositions containing said chimeric monoclonal antibodies and / or derivatives thereof.

An inverted microwell (102) provides rapid and efficient microanalysis system (100) and method for screening of biological particles (128), particularly functional analysis of cells on a single cell basis. The use of an inverted open microwell system (102) permits identification of particles, cells, and biomolecules that may be combined to produce a desired functional effect also functional screening of secreted antibody therapeutic activity as well as the potential to recover cells and fluid, and optionally expand cells, such as antibody secreting cells, within the same microwell.

The invention discloses a preparation method of an anti-bisphenol A monoclonal antibody. The preparation method comprises the following steps of: combining bisphenol A and a macromolecule carrier protein to prepare an artificial immunity antigen and a coating antigen; preparing splenocyte suspension from a bisphenol A artificial immunity antigen immunity mouse; fusing the mouse splenocyte and mouse myeloma cells SP2 / 0 in the presence of polyethylene glycol; detecting by hybridoma technology and subcloning for many times to obtain positive hybridoma cells which can stably secrete bisphenol A antibody, wherein the secreted antibody belongs to the IgG1 subclass; and establishing a bisphenol A indirect competition ELISA (Enzyme-Linked Immunosorbent Assay) test by utilizing the monoclonal antibody secreted by cell lines with high antibodytiter, wherein the minimum limit of detection can reach 0.324ng / mL. The antibody has no cross reaction with compounds with benzene ring structures, such as benzene, phenol, tert-butylphenol, p-hydroxylphengl, o-hydroxybenzoic acid and the like, has strong specificity and can be used for immunity detection of bisphenol A.

The invention provides a PED (Porcine Epedemic Diarrhea) inactivated vaccine and a preparation method thereof and relates to the field of biopharmacy. The PED inactivated vaccine comprises inactivated PEDV (Porcine Epedemic Diarrhea Virus), and is characterized in that the PED inactivated vaccine comprises 0.05-10 mg / mL Beta-glucosylceramide, 0.1-21 mg / mL monophosphoryl lipid A, 1.5-125 mg / mL muramyl dipeptide and 0.7-4.5 mg / mL Beta-glucan. According to the ingredients of the PED inactivated vaccine, Beta-glucosylceramide, monophosphoryl phosphoryl lipid A, muramyl dipeptide and Beta-glucan have the synergistic effect, the immune response of animals to antigens in the vaccine is significantly improved, the immune window phase is shortened, the antibody production duration of the animal body is obviously prolonged, the serum antibody level is improved, and the level of total intestinal mucosa secretory antibodies (the total SIgA) is improved.

The invention relates to a CD19 targeted chimeric antigen receptor and an application thereof, and particularly provides a polynucleotide sequence, which is selected from: (1) a polynucleotide sequence which contains, in a successively connected manner, an encoding sequence of anti-CD19 single-chain antibody, an encoding sequence of human CD8[alpha] hinge zone, an encoding sequence of human CD28 transmembrane zone, an encoding sequence of human CD28 intracellular zone, an encoding sequence of human CD3 [zeta] intracellular zone, and optionally, a fraction, which contains an extracellular domain III and an extracellular domain IV of EGFR, and an encoding sequence of anti-human PD1 sequence fraction; and (2) a complementary sequence of the polynucleotide sequence (1). The invention also provides related fusion proteins, a carrier containing the encoding sequences, and applications of the fusion proteins, encoding sequences and carrier. The CAR-T cell has strong killing effect on specifictumor cells, and can reach more than 90% in killing efficiency on the specific tumor cells under the multiplicity of infection of 1:2. The CAR-T cell can secrete the PD1 antibody and has regulation effect on immunosuppression micro-environments.

The present invention relates to THE archaeological detection field, and discloses a method for preparing a tussah silk fibroin protein monoclonal antibody, a FeCl2 and tetrasodium iminodisuccinate mixed solution system is used for extraction of tussah silk fibroin, the tussah silk fibroin as a complete antigen is injected into the body of rabbits, a rabbit with high immunizing potency is selected, splenic lymphocytes and myeloma cells of the rabbit with the high immunizing potency are fused, after obtained hybridoma is cultured, a bacterial strain with positive secreted antibodies, strong antibody secreting ability and good cell growth conditions can be selected by indirect ELISA method, the bacterial strain is cloned by limiting dilution method until the secreting positive rate of the antibody with grown hybridoma is 100%, the selected cells are used for large-scale production, and then a chromatographic column is used for purification of the monoclonal antibody. The antibody prepared by the method has strong specificity, and in the application process, the operation is simple and quick, and the detection accuracy is high.

The invention discloses an anti-human CEACAM5 monoclonal antibody and a preparation method and application thereof, and the anti-human CEACAM5 monoclonal antibody is produced by a mouse hybridoma cellline with the accession number CCTCC NO: C2016129. The preparation method comprises the following steps: S1, immunizing a mouse with whole cell protein, lysed by human gastric cancer cells, as an immunogen; S2, fusing spleen cells of the immunized mouse obtained in the step S1 with mouse myeloma cells; S3, selecting a hybridoma cell line stably secreting antibodies after cell cloning occurs during the cell fusing in the step 2; and S4, preparing the anti-human CEACAM5 monoclonal antibody from the hybridoma cell line stably secreting the antibodies obtained in the step 3. The monoclonal antibody can be used to prepare a composition for treating a malignant tumor. The anti-human CEACAM5 monoclonal antibody can be widely used in various routine operations such as flow cytometry, immunohistochemical staining and immunoprecipitation.

The invention relates to the field of archaeological detection and discloses a preparation method of a bombyx mori silk fibroin monoclonal antibody. The preparation method comprises the following steps: extracting bombyx mori silk fibroin by adopting an isooctane, n-octyl alcohol, bi(2-ethylhexyl) sodium sulfosuccinate and polyvinyl pyridine quaternary ammonium salt mixed system, injecting the bombyx mori silk fibroin serving as a complete antigen into the bodies of rabbits, selecting the rabbits with high immunizing potency, fusing splenic lymphocytes and myeloma cells, cultivating the obtained hybridoma cells, selecting strains with positive secretory antibodies, high secretion ability and good cell growth state by an indirect ELISA method, cloning by a limited dilution method until the antibody secretion positive rate of hybridoma cell growth is 100 percent, performing big turn production on the screened cells, and purifying the obtained monoclonal antibody by a chromatographic column. The antibody prepared by the method has high specificity, and is simple and rapid in operation during the application process and high in detection accuracy.

The invention discloses a chimeric antigen receptor CD19 scFV-CD8hinge-CD28TM-CD28-CD3zeta joint aPD1 scFV CAR-T cell and use thereof. Specifically, the invention provides a polynucleotide sequence selected from: (1) a coding sequence containing sequentially linked anti-CD19 single chain antibodies, a coding sequence of a human CD8alpha hinge region, a coding sequence of a human CD28 transmembraneregion, a coding sequence of a human CD28 intracellular region, a coding sequence of a human CD3zeta intracellular region, a coding sequence of a P2A polypeptide, a human IL2 signal peptide coding sequence, and an anti-human PD1 monoclonal antibody coding sequence; and (2) a complementary sequence of the polynucleotide sequence in (1). The invention further provides a relevant fusion protein, a vector containing the coding sequences, and use of the fusion protein, the coding sequences and the vector. A prepared CD19-28z-aPD1 CAR-T cell has a very strong killing function on specific tumor cells. The prepared CD19-28z-aPD1 CAR-T cell has the function of secreting a PD1 antibody, and can block the site of PD1-PDL1 to prevent CAR-T cells from being inhibited by PD1.

The invention relates to the identification of secretory antibody-bound bacteria in the microbiota in a subject that influence the development and progression of inflammatory diseases and disorders. Thus, the invention relates to compositions and methods for detecting and identifying the constituents of a subject's microbiota, methods of modifying the constituents of the microbiota, and methods for treating inflammatory diseases and disorders in a subject in need thereof.

The invention relates to the field of tumor immunology, in particular to construction and application of chimeric antigen receptor-T (CAR-T) cells capable of targeting mesothelin and carrying a PD-L1blocking agent. The CAR-T cells are constructed by transfecting a CAR capable of targeting the mesothelin and carrying the PD-L1 blocking agent, wherein the CAR capable of targeting the mesothelin andcarrying the PD-L1 blocking agent is formed by a leader peptide of a CD8 antigen, an anti-mesothelin single-chain antibody, a hinge region, a transmembrane region, an intracellular signal domain andan anti-PD-L1 antibody secretion region which are sequentially connected with one another. The mesothelin SS1P scFv of the CAR-T cells provided by the invention can specifically bind to tumor cells expressing mesothelin glycoprotein and promote the secretion of anti-tumor-related cytokines, thereby achieving a killing effect on the tumor cells; furthermore, the CAR-T cells provided by invention can also secrete a PD-L1 antibody, specifically bind to PD-L1 produced by tumor cell expression, and block the inhibitory effect of a PD-L1 / PD-1 signal on T cell activity, thus facilitating the long-term effective inhibition of the CAR-T on tumor cell growth.

The invention discloses a GPC3 (glypican-3) monoclonal antibody hybridoma strain 8G6, and a preparation method and application thereof, which belong to the technical field of cell engineering. The preservation code of the GPC3 monoclonal antibody hybridoma strain 8G6 is CGMCC (china general microbiological culture collection center) No.5427. The preparation method of the GPC3 monoclonal antibody hybridoma strain 8G6 includes: using GPC3 protein as immunogen to immunize a mouse; and fusing mouse splenocytes having serum titer more than 1:104 with SP2 / 0 myeloma cells, using an HATRPMI-1640 medium to screen fusion cells, and finally obtaining the hybridoma strain 8G6 by ELISA (enzyme-linked immunosorbent assay) and repeated limiting dilution. The GPC3 monoclonal antibody hybridoma strain 8G6 is high in yield of secretory antibodies, and is easy to survive, and the secreted monoclonal antibodies are high in titer, sensitive in reaction, easy to detect, low in production cost and widely applicable to detection of GPC3 protein expression.

The invention relates to an anti-H7 subtype avian influenza virus monoclonal antibody and application thereof, belonging to the technical field of biology. A hybridoma cell strain 5D2 is screened; the hemagglutination inhibition (HI) titer of the rat ascites antibody prepared from the hybridoma cell strain 5D2 for H7-AIV is 11log2, and the ELISA titer is 2*10<5>. The anti-H7 monoclonal antibody secreted by the hybridoma cell strain has high HI and ELISA titer; the colloidal gold immunochromatography detection test strip assembled by the anti-H7 monoclonal antibody has high specificity, except that the result for H7-AIV is positive, the results for the other avian viruses are negative; the anti-H7 monoclonal antibody can be used for on-site diagnosis on fowl, mammals or the like infected by H7-AIV; and the result can come out within 10 minutes, is visual, and can be easily judged with naked eyes. The sensitivity of the diagnosis kit assembled by the hybridoma cell strain secretory antibody is not lowered after the diagnosis kit is stored for one year, so the diagnosis kit has favorable stability.

The invention discloses a preparation method for a CAR-T cell targeted to EGFRvIII and modified by a gene and application. The method comprises the following steps: optimizing an amino acid sequence codon targeted to the EGFRvIII, enabling the amino acid sequence codon to be more suitably expressed in a human source host body, and preparing the CAR-T cell; knocking out a PD-1 gene in the CAR-T cell by using a single base editing method; or combining a secretion interleukin 12 (IL-12), and using for promoting proliferation of the activated T cell; or combining a secretion anti-PD1 antibody, andusing for combining with the PD1 on other T cells, and relieving the inhibiting effect of a tumor tissue to the T cell. The method is capable of improving transfection efficiency and expression efficiency of the imported gene through selecting a PiggyBac transposon system, shortening operating time, and simplifying a preparation process of the CAR-T. In addition, the method avoids using of a virus vector while knocking out the PD-1 gene, so the safety is higher.

The invention discloses a microbial feed additive capable of enhancing immunity of fish and an application of the feed additive. The microbial feed additive includes, by weight, 16-22 parts of fructus forsythiae, 16-19 parts of herba andrographitis, 12-17 parts of artemisia apiacea, 15-21 parts of Chinese rhubarb, 1-2 parts of angelica sinensis, 13-20 parts of houttuynia cordata, 10-15 parts of radix glycyrrhizae, 7-11 parts of acanthopanax roots, 0.6-0.9 parts of garlic, 2-3 parts of pericarpium citri reticulatae, 3-5 parts of eucommia ulmoides, 0.8-1.3 parts of bacillus subtilis, and 1-1.4 parts of lactobacillus plantarum. The microbial feed additive is free of toxic and side effects, has no pollution and residual, is free of generating drug resistance, can promote secretion of antibodies by a host, improves activity of immune cells of the host and improves immunity and resistance of body. The microbial feed additive also maintains microecological balance in intestinal tracts and best immune state, and also improves digestion and absorption rate of nutrients.

The invention discloses a preparation method of an alpha-glucan antigen and a specificity monoclonal antibody. The preparation method of the antigen comprises the following steps: mixing alpha-glucan and protein; dissolving with water; regulating the pH of solution to 5-8; carrying out freeze drying, and grinding into powder; putting the powder in the environment with constant temperature of 50-60DEG C and relative humidity of 60-90%; reacting for at least three days to obtain the alpha-glucan antigen. The preparation method of the antibody comprises the following steps: mixing the prepared antigen with an adjuvant, emulsifying the mixture, and injecting the emulsified mixture into animal bodies; blending the splenocyte and oncocyte of the animal body with the high serum valence to obtain a hybridoma cell capable of secreting the antibody; and producing the antibody by the hybridoma cell or with an ascites method. The antibody prepared by the method has high immunogenicity, and can be used for effectively inducing to generate immune response to obtain the antibody with the excellent property. The antibody obtained by the method provided by the invention can be subjected to specificity reaction with the alpha-glucan to generate macroscopic precipitates, and the antibody has high valence and can be used for the quantitative analysis on the alpha-glucan.

The present invention relates to expression and assembly of foreign multimeric proteins—e.g., antibodies—in plants, as well as to transgenic plants that express such proteins. In one of several preferred embodiments, the generation and assembly of functional secretory antibodies in plants is disclosed. The invention also discloses compositions produced by the transgenic plants of the present invention and methods of using same.

The invention discloses an antigen for preparing a listeria monocytogenes monoclonal antibody. An amino acid sequence of the antigen is shown in SEQ NO.1. According to the antigen, a specific surfaceprotein fragment capable of representing listeria monocytogenes is obtained through screening and experimental verification, the specific surface protein fragment is used as an antigen sequence to obtain a positive cell line by cell fusion, after three rounds of subcloning, an indirect ELISA method is used to detect the positive strains and detect a cross-reaction between a secreted antibody and other food-borne pathogenic strains to obtain a specific fusion cell line, finally, a monoclonal hybridoma cell with high titer and good specificity is obtained, and the mouse monoclonal antibody is prepared and has a titer of 810000 and good specificity. The monoclonal antibody provided by the invention has higher sensitivity, good specificity and strong stability.

The invention discloses a preparation method of a nanobody. The preparation method comprises the following steps: immunizing animals with a target antigen to obtain peripheral blood mononuclear cellsof the immunized animals, and enriching B cells; marking the target antigen by using FITC fluorescence; culturing the cells by using a semi-solid culture medium to form independent single B cell clones, selecting single B cell clones secreting positive heavy-chain antibodies, extracting total RNAs of positive B cells, carrying out reverse transcription to obtain cDNAs, and carrying out VHH fragment amplification and sequencing; and performing recombinant expression and activity identification of the nanobody. According to the method, the positive B cell clones secreting VHH antibodies can be directly observed and selected, and fluorescence signals are formed through the secreted antibodies, so the secretion functional characteristics of B cell antibodies are better conformed to. The nanobody screened and prepared by the method has the advantages of small molecular weight, strong stability, high affinity, good solubility, easiness in expression and the like, and has a wide application scope.

The invention relates to a rabies virus monoclonal antibody with neutralization activity and capability of specifically recognizing rabies virus glycoprotein G. The monoclonal antibody is authenticated to be IgG1-subtype. According to the rabies virus monoclonal antibody disclosed by the invention, rabies virus inactivation totivirus is taken as an immunogen for immunizing a mouse, an antibody with neutralization activity is screened, the neutralization titer is high, the secretion of hybridoma for the antibody is stable, and the antibody is easy to store; and an in-vitro preparation link for glycoprotein is avoided, and innovativeness is achieved. A colloidal gold test paper card or test paper strip prepared from the antibody is high in test sensitivity and capable of achieving 0.03 IU / mL, capable of being used for test on human rabies virus or quality control in a vaccine production process, and high in application value.

A monoclonal secretory IgA antibody, which binds to and neutralizes human TNFα. The secretory antibody is useful in treating a variety of inflammatory conditions in humans.

The invention discloses a preparation method and application of gene-edited CAR (Chimeric Antigen Receptor)-T cells capable of secreting a cell factor or an antibody. The method for preparing the CAR-T cells comprises the step of transfecting plasmid vectors containing gene-edited plasmids and CAR modification to T cells. The CAR-T cells provided by the invention are used for knocking out negativeregulator genes on surfaces of the T cells through gene editing and gene modification, integrating a single-chain antibody for identifying a tumor cell surface antigen and an expression frame for secreting the cell factor or secreting the antibody into the T cells and continuously expressing in the T cells, so that the T cells returned back are prevented from being inhibited by a tumor microenvironment and a better killing effect on the tumor cells is realized.

The invention discloses an anti-hepatitis B monomer clone antibody hydridoma series construction method that adopts EBV transforming human B lymphocyte to gain transforming B lymphocyte cell series and taking cell melting with rat plasmocytoma cell series FO to construct excreting anti-HBshMcAb hydridoma series. The invention is easy to cultivate and has high melting. It could be used in cure lymphocyte B, HB advanced stage liver cancer.

A monoclonal secretory IgA antibody, which binds to and neutralizes human p40 (the p40 subunit of IL-12 and IL-23). The secretory antibody is useful in treating a variety of inflammatory conditions in humans.

The invention belongs to the field of biological products, and in particular relates to an inactivated vaccine for porcine epidemic diarrhea virus which is produced by ST cells and a preparation method of the inactivated vaccine. The method comprises four steps, namely amplification culture of the ST cells, virus inoculation, multiplying of vaccine producing virus liquid, and product preparation. The vaccine, which is produced by the method, is high in immunization level and short in immunological window phase; an immunity period is obviously prolonged; and total secretory antibody (total SIgA) in intestinal mucosa is remarkably improved.

A recombinant expression vector capable of expressing and secreting an antibody fragment fused with E. coli thermostable enterotoxin signal sequence derivative or E. coli outer membrane protein A (Omp A) signal sequence in the form of a soluble heterozygote protein is used to mass-produce the antibody fragment by culturing a microorganism transformed with the expression vector in a medium and collecting the antibody fragment secreted from the transformed microorganism into the medium

The invention discloses a secretory target Lewis-Y CAR-T cell, which comprises a CAR-T cell which targets Lewis-Y and secretes IL12 cytokine; a CAR-T cell which targets Lewis-Y and secretes PD-1 antibodies. The invention further discloses a preparation method of the CAR-T cell of the secretory target Lewis-Y and an application of the CAR-T cell in preparing an anti-tumor cell therapy medicament. The PD-1 antibody secreted by the CAR-T cell is specifically combined with the PD-1 on the surface of the T cell, thereby blocking the inhibition reaction of the PD-1 / PD-L1 signal on the activity of the T cell, so that the effective inhibition of growth of tumor cells for a long time by the CAR-T cell is facilitated; the IL12 cell factor secreted by the CAR-T cell can act on immunosuppressive cellsin tumor inhibitory immune microenvironment, so that the immunosuppressive capacity of the immunosuppressive cells is reduced, and the killing effect of the CAR-T cell is further enhanced.

The present invention relates to expression and assembly of foreign multimeric proteins—e.g., antibodies—in plants, as well as to transgenic plants that express such proteins. In one of several preferred embodiments, the generation and assembly of functional secretory antibodies in plants is disclosed. The invention also discloses compositions produced by the transgenic plants of the present invention and methods of using same.