43 results about "Antibody secretion" patented technology

A cell surface molecule that is expressed specifically in thymocytes, lymphocytes activated by ConA-stimulation, and peripheral blood lymphocytes. This molecule is involved in signal transmission of the secondary signal (costimulatory signal) essential for the activation of lymphocytes such as T cells and regulates functions of activated lymphocytes such as activated T cells. Disclosed are an antibody or a portion thereof, which binds to a polypeptide of the cell surface molecule, a polypeptide fragment thereof, or a fusion polypeptide comprising the fragment; a cell secreting the antibody or its portion; a pharmaceutical composition comprising the antibody; and methods of using the compositions for therapeutic, diagnostic and / or experimental purpose.

The invention provides a cell microporous chip which can detect a cell secreting a specific antibody in high throughput at the single cell level. The chip is suitable for identifying and analyzing the specific antibody secretion conditions of B cells (from people or other vertebrates), hybridomas (from mice or people) or engineering cells secreting antibodies at the single cell level and for finding out the single antibody secreting cell with good specificity, high affinity and high secretion level, is convenient to prepare, is economical and is simple to operate.

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 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 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 relates to a method for detecting a monoclonal antibody and application thereof, and the method for detecting the monoclonal antibody comprises the following steps: acquiring hyperspectral data of a cell to be detected through hyperspectral imaging detection, and inputting the hyperspectral data into a machine learning model to obtain an antibody secretion type of the cell to be detected. By introducing a hyperspectral imaging technology and a microfluidic technology, single cell separation is realized, a hyperspectral image of a single cell is rapidly acquired, required featureinformation is rapidly and accurately mined from a large amount of hyperspectral data by using a machine learning algorithm, and high-precision identification and classification of single cells are automatically realized. The analysis process has the characteristics of high flux and intelligence, high result accuracy, and high sensitivity, and a rapid, non-contact and damage-free identification method is provided for antibody drug discovery and cell analysis.

The present invention relates to a pharmaceutical composition, food composition and cosmetic composition for preventing, treating, or improving allergic diseases comprising extract or fraction of a plant of the Justicia genus as an active ingredient. The extract or a fraction of a plant of the Justicia genus according to the present invention can inhibit IgE antibody secretion and the degranulation of mast cells and basophils, and exhibits an excellent anti-allergic effect, and thus can effectively prevent, treat, or improve allergic diseases.

The invention proposes PD-L1 antibody secretion anti-mesothelin CAR-T cell tumor immunotherapy. CAR-T cells co-express an anti-PD-L1 fusion antibody, non-functional EGFR and a chimeric antigen receptor, wherein the anti-PD-L1 fusion antibody is formed by linking an anti-PD-L1 single-chain antibody to an IgG1Fc fragment containing amino acid mutation. The transgenic lymphocyte secretes the anti-PD-L1 single-chain antibody and an IgG Fc fusion antibody, and has the characteristics of resisting tumor cell-mediated immunosuppression, significantly enhanced killing ability to tumor cells, especially a significant targeted killing effect on tumors highly expressing mesothelin and PD-L1 molecules, and high safety.

The invention relates to a diluent for a hog cholera live vaccine (spleen and lymph tissue origin), belonging to the technical field of veterinary biological products. Every 1000ml of phosphate buffer solution of the diluent contains 15-50g of Chinese medicine polysaccharide, 20-60g of Chinese medicine flavone, 5-10g of vitamin and 5-40mg of bursin, wherein the Chinese medicine polysaccharide and the Chinese medicine flavone are extracted from the following Chinese medicine compositions: fructus forsythia, bistort, cyrtomium fortunei, cirsium japonicum, sanguisorba officinalis, subprostrate sophora, antelope's horn and Chinese violet. After an organism is immunized by the vaccine prepared from the diluent, differentiation and proliferation of a B lymphocyte precursor of the organism can be facilitated, the speed of transcribing DNA (deoxyribonucleic acid) into mRNA (Messenger Ribonucleic Acid) in B lymphocyte is accelerated, the generation of proteins in the B lymphocyte is promoted, and thus the generation and antibody secretion capabilities of the B lymphocyte are enhanced, and the immunity of the organism can be improved.

The present invention provides methods of quantitating recent secreted antigen specific antibodies from supernatant of antibody secreting cells (ASC) in vitro culture for evaluation of vaccine or antigen induced antigen specific antibody secretion without ex vivo antigen stimulation.

It is an object of the present invention to provide signal sequence information capable of secreting an antibody to the outside of cells in generation of the antibody by microorganisms of genus Bifidobacterium, and an antibody expression vector capable of secreting an antibody to the outside of cells by utilizing the signal sequence information. As a means for achieving the aforementioned object, there is prepared Bifidobacterium longum, which is transformed with a vector having inserted thereinto a DNA insert comprising the 5′-terminus of an antibody gene linked to the 3′-terminus of a DNA encoding a signal peptide-linker conjugate having a linker linked to the C-terminus of a signal peptide consisting of an amino acid sequence shown in SEQ ID NO: 1.

The invention relates to a freeze-drying protective agent for a hog cholera live vaccine (spleen and lymph tissue origin), belonging to the technical field of veterinary biological products. Every 1000ml of phosphate buffer solution of the freeze-drying protective agent contains 30-100g of Chinese medicine polysaccharide, 20-60g of Chinese medicine flavone and 2-16mg of bursin, wherein the Chinese medicine polysaccharide and the Chinese medicine flavone are extracted from the following Chinese medicine compositions: sedum sarmentosum, radix tinosporae, Chinese fevervine, buffalo horn, mole cricket, elsholtzia, radix stephaniae tetrandrae, Chinese lobelia and beautiful sweetgum fruit. After an organism is immunized by the vaccine prepared from the freeze-drying protective agent, differentiation and proliferation of a B lymphocyte precursor of the organism can be facilitated, the speed of transcribing DNA (deoxyribonucleic acid) into mRNA (Messenger Ribonucleic Acid) in B lymphocyte is accelerated, the generation of proteins in the B lymphocyte is promoted, and thus the generation and antibody secretion capabilities of the B lymphocyte are enhanced, and the immunity of the organism can be improved.

The invention provides polyester fiber containing a wormwood component and wormwood fragrance and a preparation method of the polyester fiber, and relates to the technical field of textile fibers. The polyester fiber containing the wormwood component and the wormwood fragrance is prepared from the following materials in parts by weight of 8 to 11 parts of radix stemonae, 6 to 9 parts of flos caryophylli, 6 to 9 parts of herba taraxaci, 8 to 11 parts of radix et rhizoma rhei, 6 to 9 parts of flos gardeniae, 2 to 3 parts of rhizoma coptidis, 30 to 35 parts of wormwood, 5 to 7 parts of cellulase, 50 to 55 parts of polyester, 15 to 18 parts of lignans, 16 to 19 parts of nanoscale titanium dioxide and 25 to 28 parts of fiber-forming polymer. By adding a wormwood powder mixture and a wormwood extract, the taste of an industrial additive is effectively removed, meanwhile, the wormwood has the mosquito-repelling and insect-preventing effects, and secondly, the fragrance generated by the wormwood acts on nasal mucosa, so that the content of immune globulin S (SlGA) secreted by antibodies on the nasal mucosa is increased; and the increase of the content enables virus not to survive easily on the nasal mucosa and the respiratory mucosa.

The invention discloses a chimeric antigen receptor targeting HER2 and an expression vector and application thereof, and belongs to the field of tumor immune drugs. The chimeric antigen receptor targeting HER2 includesa signal peptide, a single-chain antibody targeting HER2, an extended CD8 alpha hinge region, a transmembrane region, an inducible co-stimulater, an intracellular signal peptide, a P2A linker peptide, an IL2 signal peptide, and an anti-PD1 single-chain antibody secretion region, wherein the signal peptide, the single-chain antibody targeting HER2, the extended CD8 alpha hinge region, the transmembrane region, the inducible co-stimulater, the intracellular signal peptide, the P2A linker peptide, the IL2 signal peptide, and the anti-PD1 single-chain antibody secretion region are connected in sequence, the nucleotide sequence of the signal peptide is shown as SEQ ID NO:1in a sequence table, the nucleotide sequence of the single-chain antibody targeting HER2 is shown as SEQ ID NO:2in the sequence table, the nucleotide sequence of the extended CD8 alpha hinge region is shown as SEQ ID NO:3in the sequence table, the nucleotide sequence of the transmembrane region is shown as SEQ ID NO:4in the sequence table,the nucleotide sequence of the intracellular signal peptide is shown as SEQ ID NO:5in the sequence table, the nucleotide sequence of the P2A linker peptide is shownas SEQ ID NO:6 in the sequence table, and the nucleotide sequence of the IL2 signal peptide is shown as SEQ ID NO:7in the sequence table. The chimeric antigen receptor has high antitumor activity while producing fewer cytokines.

The invention provides a method for generating a novel induction antibody, which comprises a step of preparing a eukaryotic expression vector expressing fusion antigen protein. The fusion protein consists of four parts of sequences: (1) a signal peptide sequence from a human IgE heavy-chain coded sequence; (2) an antigen protein sequence; (3) an auxiliary sequence from a mycobacterium tuberculosis cytochrome C oxidase subunit II, wherein the sequence can amplify the antibody response of the rat and mice against the antigen protein; and (4) a polypeptide sequence MFSRMTSLIMGN that can be combined with the mice FcrR II (APCTS for short), wherein the sequence can specifically guide the protein antigen to the mice antigen-presenting cell (APC) so as to improve the antigen presenting efficiency. The plasmid expressing the fusion antigen protein is directly injected into the mice muscle or skin, and the mice cells can take in DNA (deoxyribonucleic acid) and the efficiently-expressed fusion antigen protein; and then the animal is stimulated to generate a high-affinity antibody against the antigen protein. The generated antibody after being purified can be applied to the scientific research and clinical diagnosis or treatment; and the antibody secretion cell can be used for preparing a specific monoclonal antibody by use of the hybridoma technology.

The invention provides a CD127 resisting antibody, a cell strain for secreting the antibody and a preparation method and application of the antibody, and belongs to the technical fields of biopharmaceutics and immunochemistry. The antibody or an antigen binding fragment of the antibody is specially bound with CD127 and can specially recognize epitope sequences of CD127. The prepared hCD127 resisting monoclonal antibody and the monoclonal antibody hybridoma cell strain CD127-7E7 for secreting the monoclonal antibody can specifically recognize human CD127, have favorable affinity, can be used for qualitative or quantitative detection of CD127 protein, and besides, can also be used for judging the epitope or functional domain of the CD127 protein, so that the monoclonal antibody and the cellstrain have favorable practical application value.

The invention belongs to the technical field of biology, and discloses a hybridoma cell strain secreting a porcine sapelovirus VP1 protein monoclonal antibody. The preservation number of the hybridoma cell strain is CCTCC NO: C2021177, and the hybridoma cell strain is preserved in China Center for Type Culture Collection, Wuhan University, Wuhan, China. The hybridoma cell has stable antibody secretion capacity, the secreted monoclonal antibody has good reaction specificity with the porcine sapelovirus VP1 protein, the antigen epitope recognized by the monoclonal antibody is the 40th-46th amino acid of the porcine sapelovirus VP1 protein, the polypeptide sequence of the antigen epitope is 40PALTAAE46, and the epitope is not reported yet. The hybridoma cell strain lays a good material foundation for researching the etiology and pathogenesis of the porcine sapelovirus in the future.

The invention discloses a method for preparing a monoclonal antibody through rapid immunization, which belongs to the technical field of diagnosis, and comprises the following steps: S1, taking animals which are not less than 7 weeks old from cultured animals for cage separation, S2, anesthetizing the animals by using 4% chloral hydrate according to the amount of 0.1 ml / 10g, S3, extracting a diluted antigen solution by using a 1ml disposable sterile syringe, and S4, fixing the narcotized animal on an operating table in a clean environment, exposing the abdomen, and removing hairs from the abdomen; S5, cutting off the outer epidermis of the abdomen by using a pair of surgical curved scissors, pushing aside adipose tissues, slightly pulling out the spleen by using tweezers, and injecting anantigen into the spleen; S6, suturing the wound by using a suture needle; S7, carrying out second and third immunization every 3-14 days. Blood is collected 25 days after the third immunization to verify the antibody secretion condition of the sensitized B lymphocytes; the invention has the advantages of reasonable and practical design, low antigen usage amount, reduced cost, improved antibody titer, and elimination of false positive to some extent. The immune operation is simple, and the situation of unsuccessful immunization caused by immunization to the subcutaneous part does not need to beworried.

The invention provides a trichina 7TR protein human single-chain antibody and a preparation method. The trichina 7TR protein human single-chain antibody is obtained through carrying out prokaryotic expression on a trichina 7TR single-chain antibody and carrying out purification and renaturation; screening a Tomlinson human single-chain antibody library; and carrying out inducible expression on a screened positive single-chain antibody secretion strain and purifying by adopting a saturated ammonium sulfate precipitation method and Protein A affinity chromatography. The single-chain antibody can be used for preparation of anti-lung cancer drugs.

The various embodiments of the disclosure relate generally to processes, methods, and systems for generating functional B cells ex vivo. It is particularly useful for ex vivo generation of antigen-specific germinal-center (GC) like B cells that are capable of efficient B cell expansion, immunoglobulin (Ig) class switching / class switching recombination (CSR), expression of germinal B cell phenotypes, antibody secretion, and somatic hypermutation (SHM) and resulting affinity maturation center phenotypes.

The invention relates to the technical field of molecular biology, and particularly relates to a signal peptide for improving antibody yield. The amino acid sequence of the signal peptide is MNFGLSWVFLVLVLKGVQC or MVFTPQILGLMLFWISASRG, or the amino acid sequence of the signal peptide is from a heavy chain and a light chain of an antibody 1G11, the signal peptide is used for improving secretory expression of target protein, and the signal peptide is secretory expression of the antibody. According to the signal peptide, PCR amplification is conducted on the signal peptide to obtain different gene segments, gel recovery is conducted, and the yield of target genes, especially antibody secretion can be improved by identifying a vector through restriction endonuclease.

The invention relates to the technical field of regulation of secretion of virus specific antibodies, and discloses 30 rats with the weight of 250-300g, half female rats and half male rats, the rats are randomly divided into three groups in a layered manner according to the weight, the three groups are subjected to experimental operation respectively, and sample collection and detection are performed after re-culture. According to an animal experiment operation method for improving antibody secretion of a novel coronavirus vaccine to special crowds, autologous lymphocytes are subjected to in-vitro induction and then are fed back; data can be obtained in an experiment to show that the antibody level of the autologous feedback of the in-vitro induced lymphocytes is in jumping increase; the experimental data can show that the operation method can overcome the defect that the elderly and the weak and sick people respond slowly or do not respond to vaccines; the specific virus immune clearance effect is improved; therefore, the occurrence rate of severe cases is reduced; and the method has a certain treatment effect on the severe cases.

The invention discloses a method for preparing a feline distemper virus monoclonal antibody by using a bioreactor. The method comprises the following steps: step 1, carrying out subcloning on hybridoma cells by using a limited dilution method, selecting single cells into a serum-free culture medium, and carrying out suspension culture to obtain a cell strain A805 with high survival rate and strong antibody secretion ability, wherein the preservation number of the strain is CGMCC No.21902; step 2, preparing anti-feline distemper virus monoclone by using a bioreactor, wherein hybridoma cells are cultured in two stages, namely a cell growth stage and an antibody secretion stage, in the bioreactor; and step 3, harvesting the cell culture fluid as an antibody, and centrifuging to remove cell debris, thereby obtaining a finished product of the feline distemper virus monoclonal antibody. According to the method, the titer of the distemper virus monoclonal antibody is improved, the defects of a traditional ascites production antibody are overcome, and the antibody product is free of impure protein, stable in physicochemical property and small in batch-to-batch difference.

A method of inducing the sustained release of antibodies in milk comprising the step of: a) implanting at least one antigen releasing device adjacent to, within close proximity of or within at least one supramammary lymph node, wherein in use the antigen releasing device releases an antigen into the tissue area around the supramammary lymph node which stimulates antibody secretion into a mammary gland.

The invention discloses a chimeric antigen receptor targeting HER2, its expression carrier and application, and belongs to the field of tumor immune drugs. Chimeric antigen receptors targeting HER2 include: sequentially linked signal peptides, single-chain antibodies targeting HER2, extended CD8α hinge region, transmembrane region, co-stimulatory factors, intracellular signal peptide, P2A linking peptide, IL2 signal peptide and the anti-PD1 single-chain antibody secretion region, the nucleotide sequence of the signal peptide is shown in SEQ ID NO: 1 in the sequence listing, and the nucleotide sequence of the HER2-targeting single-chain antibody is shown in SEQ ID NO: 2 in the sequence listing The nucleotide sequence of the extended CD8α hinge region is shown in SEQ ID NO: 3 in the sequence listing, the nucleotide sequence of the transmembrane region is shown in SEQ ID NO: 4 in the sequence listing, and the nucleotide sequence of the intracellular signal peptide The acid sequence is shown in SEQ ID NO: 5 in the sequence listing, the nucleotide sequence of the P2A connecting peptide is shown in SEQ ID NO: 6 in the sequence listing, and the nucleotide sequence of the IL2 signal peptide is shown in SEQ ID NO in the sequence listing: 7. The chimeric antigen receptor produces fewer cytokines while possessing high antitumor activity.

The invention discloses a construction method and application of peripheral blood B cell lines (BCL). The BCL constructed in the invention have the characteristic of immortalized growth, the proportion of CD19<+> cells in the BCL is increased, the expression levels of CD138, CD38 and CD27 in the CD19<+> BCL are significantly higher than that of CD19<+> B cells before Epstein-Barr virus (EBV) transformation, and the capacities of antibody secretion and antigen presentation of the B cells are retained. The BCL disclosed by the invention can be used for screening dominant T cell immune response epitopes, or preparing a high-titer anti-HBsAg monoclonal antibody and inducing an HBV specific T cell line, and providing a new direction for removing hepatitis B.

The method for rapidly detecting the antibody secreted by the hybridoma cell comprises the following steps: displaying an Fc segment of the antibody in the hybridoma cell in cytoplasm, analyzing the Fc segment in the cytoplasm, and judging the antibody specificity and the antibody generation capability of the cell. According to the method provided by the invention, the positive rate proportion of antibody cells secreting antibodies in some intracellular expression antibodies and hybridoma cells in the total cell number can be visually detected. The antibody secretion capability of each monoclonal cell and the non-specific component on the surface of the hybridoma cell can be detected.