217 results about "Cell Clone" patented technology

A method for identifying and isolating cells which produce secreted proteins. This method is based upon a specific characteristic or the expression level of the secreted protein by transiently capturing the secreted protein on the surface of an individual cell, allowing selection of rare cell clones from a heterogeneous population. Also provided is the use of this method to generate cells which produce a desired level of secreted protein or secreted protein of a particular characteristic(s), and organisms which possess such cells. In particular, the method allows rapid isolation of high expression recombinant antibody-producing cell lines, or may be applied directly to rapid isolation of specific hybridomas, or to the isolation of antibody-producing transgenic animals. This method is applicable for any cell which secretes protein.

The present invention provides a method of carrying out adoptive immunotherapy in a primate subject in need thereof by administering the subject a cytotoxic T lymphocytes (CTL) preparation in a treatment-effective amount. The method comprises administering as the CTL preparation a preparation consisting essentially of an in vitro expanded primate CTL population, the CTL population enriched prior to expansion for central memory T lymphocytes, and depleted prior to expansion of effector memory T lymphocytes. In some embodiments, the method may further comprise concurrently administering Interleukin-15 to the subject in an amount effective to increase the proliferation of the central memory T cells in the subject. Pharmaceutical formulations produced by the method, and methods of using the same, are also described.

The invention provides methods for producing mixtures of antibodies from a single host cell clone, wherein, a nucleic acid sequence encoding a light chain and nucleic acid sequences encoding different heavy chains are expressed in a recombinant host cell. The recombinantly produced antibodies in the mixtures according to the invention suitably comprise identical light chains paired to different heavy chains capable of pairing to the light chain, thereby forming functional antigen-binding domains. Mixtures of the recombinantly produced antibodies are also provided by the invention. Such mixtures can be used in a variety of fields.

Methods are provided for the detection and analysis of clonality in a cell population, where parallel sequencing is applied to a nucleic acid sample obtained from the cell population, optionally a population of lymphocytes. Replicate samples are amplified, and sequenced, where identification of coincident sequences in two or more replicates is indicative of clonal expansion.

The invention relates to a real-time path planning method of AUV (Autonomous Underwater Vehicle), in particular to a method for carrying out online, real-time local path planning according to an online map in an AUV real-time collision preventation process. The method comprises the steps of: setting the quantity of small populations according to the quantity of path points of the AUV, initializing; carrying out immune selection on each small population to obtain subgroups; carrying out genetic manipulation on one subgroup, carrying out cell cloning on the other subgroup; then clustering through a vaccination and an antibody to form the next generation of small population, judging whether the next generation of small population meets the conditions or not; if yes, selecting optimal individuals of the small populations; and selecting the optimal individuals from the set consisting of all optimal individuals to be used as a planning path. According to the invention, the diversity of the population is maintained by using an antibody clustering principle, the premature convergence of an algorithm is avoided, and the global optimization is facilitated. The established immune genetic algorithm is used for clustering and analyzing generated filial generations by adopting a self-regulating mechanism, and the diversity of the population is ensured.

This invention relates to pork liver cell immortalities and its producing method. Procedures are showed: high live ratio fresh original pork liver cell is got by dispase-collagenase perfusion method; it is cultured for 24 hours. Then cell upper heat removing of recombination retronituse that contains SV40 big T antigen is used to infect the pork cell under condition of polybrene which concentration is 8ug / ml. Then it is medicine pressure filtrated by 500ug / ml G418 after one week, single clone cell is picked for large culture when the cell clone appears and grows to 1.0-2.0cm to get pork cell that can passage. Reinfection is done under condition of 8ug / ml polybrene. Then it is medicine pressure filtrated by 2ug / ml puromycin after one week. Single clone cell is picked for large culture when the cell clone appears and grows to 1.0-2.0cm to get immortality pork cell.

A process for screening an expression library to identify clones expressing enzymes having a desired activity is provided. The process involves first generating from genomic DNA samples of one or more microorganisms an expression library comprising a plurality of recombinant cell clones, and then introducing into capillaries in a capillary array a substrate and at least a subset of the clones, either individually or as a mixture. Interaction of the substrate and a clone expressing an enzyme having the desired activity produces an optically detectable signal, which can then be spatially detected to identify capillaries containing clones producing such a signal. The signal-producing clones can then be recovered from the identified capillaries.

An interleukin-15 (IL-15) gene modified natural killing cell strain for immunotherapy of tumor is prepared through inserting the cDNA coding region of IL-15 in pcDNA3 eucaryotic expression carrier, configuring secretion-type recombinant eucaryotic expression carrier pc DNA3 / IL-15, using liposom to transfecte cell NK-92, adding Geneticin (G418) to screening culture medium, screening, limited dilution to cloned cells, detecting activity, choosing positive cell clones, and amplification.

The invention discloses a culture medium for establishing a pig iPS cell line and a culture method thereof. A typical pig iPS cell is obtained in the ninth day through transfection by four transcription factors of OCT4, SOX2, KLF4 and c-MYC and induction culture of the culture medium, and the pig iPS cell can be obtained efficiently. The obtained pig iPS cell clone is flat clone, has a regular edge and is similar to the ES cellular morphology of the human body. According to the pig iPS cell line, the pig iPS cell through subculture keeps the undifferentiated state, shows positive in the AP dyeing displaying result and has a pluripotency mark, and the differentiated cell in vitro expresses NCSTN (entoderm), NESTIN (ectoderm) and DESMIN (mesoblast).

The invention provides a preparation method and application of a product obtained by coupling penicillin with carrier protein, as well as a beta-lactam type penicillin antibody. Animals are immunized with penicillin artificial antigen coupled in the invention so as to prepare the antibody which can be used for detecting beta-lactam type penicillin in foods. The preparation method comprises the following steps: immune BALB / C mouse spleen cells and SP2 / 0 mouse myeloma cells are fused; beta-lactam type antibiotics coupled with the carrier protein are used as coating antigen to screen positive hybridoma; hybridoma capable of stably transferring culture and secreting anti-beta-lactam type antibiotic antibodies through cell clones is obtained; and an ascites monoclonal antibody is prepared. The prepared monoclonal antibody is utilized to establish a direct competitive ELISA method having high specificity, sensitivity and accuracy to the beta-lactam type antibiotics, as well as an immune colloidal gold test strip. The preparation method for the product obtained by coupling penicillin with carrier protein, as well as the beta-lactam type antibiotic antibodies can serve the rapid detection of beta-lactam type antibiotic residue in foods.

The invention discloses an efficient genetic-modification-free iPSC induced and industrialized monoclonal picking platform and application. The platform can efficiently perform reprogramming and only needs to use a minimum number of reprogramming factors (OCT4, SOX2, E6 and E7). In a monoclonal separation stage, SSEA4 / TRA-1-60 is taken as a selection marker, a large number of single cell clones are obtained through a flow cytometry sorting technology. The platform has the advantages of being high in reprogramming efficiency, high in safety, easy to operate, capable of achieving large-scale production and the like.

The invention relates to the field of biotechnology, and particularly relates to a method for performing screening in virus-sensitive cell clonal strains by applying an indirect immunofluorescence assay technology. The method comprises the following steps of: identifying the purity of a cell line; obtaining monoclonal cell strains in the cell line; and identifying the sensitivity of the monoclonal cell strains to virus. The method disclosed by the invention has the beneficial effects of being short in detection time, capable of identifying the sensitivity of single-cell cloning to virus only by 3 days, relative simple to operate, easy, capable of being used for screening lots of non-cytopathic virus-sensitive cell strains, stable in result, strong in repeatability, and consistent with the effect of a virulence determination method.

The invention relates to a hoofed mammal inducible multipotential stem cell and a preparation method thereof. The method comprises the following steps of: A) constructing an expression vector carrying a transcription factor, wherein the transcription factor is Oct4, Sox2, c-Myc, Klf4, Lin28 and Nanog; and B) introducing the transcription factor in the step A) into the cells of the hoofed mammal in a combining form; picking clones of which the form is similar to that of the embryonic stem cell for subculturing; and screening the cell clones meeting the characteristic of the embryonic stem cell to obtain the hoofed mammal iPS cell. The method contributes to determining the most proper culturing condition and the most proper culturing method for establishing an ES cell line of pig, sheep, cattle and other hoofed mammals; and experimental models for various genetic diseases of human beings can be established with the hoofed mammal iPS cell line.

A method of producing ethanol, which comprises of starch obtained from continuously cultivated unicellular green algae strains which reproduce through a single cell clone cultivation method that cyclically produces starch extra-cellularly. Only the starch is recovered and goes through a saccharification and fermentation process for the production of ethanol. The algae are left for continual reprocessing. The obtained starch is then saccharified and fermented to produce ethanol. This production method is available and feasible in any part the world, from tropical areas to high latitude areas because it is controlled not by natural climatic conditions but in an environment that is monitored and controlled by humans. The continuous production process from these Chlorella algae strains can reduce the production cost of ethanol production as well as contribute to reducing industrial wastes and carbon dioxide to contribute to the earth's environmental wellbeing.

The present invention discloses a porcine circovirus type II strain, wherein the latin name is Circoviridae, and the preservation number is CCTCC NO:V201312. According to the invention, in vitro PCV2 culture proliferation conditions are optimized, and a PK15 cell clone strain with high PCV2 sensitivity and named L8 is screened so as to establish a foundation for further PCV2 researches.

A method of producing ethanol, which comprises of starch obtained from continuously cultivated unicellular green algae strains which reproduce through a single cell clone cultivation method that cyclically produces starch extra-cellularly. Only the starch is recovered and goes through a saccharification and fermentation process for the production of ethanol. The algae are left for continual reprocessing. The obtained starch is then saccharified and fermented to produce ethanol. This production method is available and feasible in any part the world, from tropical areas to high latitude areas because it is controlled not by natural climatic conditions but in an environment that is monitored and controlled by humans. The continuous production process from these Chlorella algae strains can reduce the production cost of ethanol production as well as contribute to reducing industrial wastes and carbon dioxide to contribute to the earth's environmental wellbeing.

The invention discloses a method for improving expression of extraneous genes by insect transgene cells. In the method, active promoter controlled neomycin resistance gene expression cassettes, enhancer elements and extraneous gene expression cassettes inside the cells of insects are cloned into a vector with reporter genes based on transposon piggyBAC factors; subsequently, the vector is mixed with subsidiary plasmids expressing transposase to transfect insect cell lines; transgene insect cells are acquired through sectionalized screening of G418. Engineering cells expressing extraneous genes at high level are acquired by cell clone technology in combination practical examination of expression level of extraneous genes. With the method, transgene insect cells can express extraneous genes at high level continuously; the expression products are free of rhabdoviruses with good bio-safety, and are processed perfectly as well as natural.

The invention provides a method for producing induced multipotential stem cell, comprising steps of: A) constructing viral vectors carrying transcription factors, wherein the transcription factors are selected from: Oct4, Sox2, c-Myc, Klf4, Lin28 and Nanog; B) respectively transfecting viral vectors obtained in step A), and obtaining virus liquid containing the transcription factors; and C) infecting cells in combination manner by the virus liquid containing the transcription factors obtained from step B)selecting clones with human embryo like stem cell for subculturing, and obtaining induced multipotential stem cell by screening cell clone according to human embryo stem cell characteristic. The method of the invention for preparing human embryo stem cell has high efficiency, can avoid rejection reaction, differentiates to be different histiocytes in specific condition, and has an extensive application future.

The present invention relates to recombinant hepatitis C virus (HCV)-derived nucleic acids and to stable rapidly growing cell clones derived from human hepatoma Huh-7 cell line and supporting high titer replication of said recombinant HCV nucleic acids. The subgenomic HCV replicons and cell clones of the instant invention represent the in vitro system of choice for studies of HCV propagation, anti-viral drug screening, and vaccine development.

A new method for selecting clones and recloning mammalian cells which are of importance for the production of biopharmaceuticals, preferably hamster or mouse myeloma cells, with a high degree of automation and throughput. The invention relates to methods of depositing and replicating single cell clones of the cells in question. The invention also relates to methods of preparing proteins using cells which have been obtained and replicated by single cell deposition as well as compositions which allow the replication of single cells.

The invention discloses a single cell cloning culture method. The single cell cloning culture method comprises the steps: step one, preparing culture solution microdrops; step two, preparing a micromanipulation system; step three, screening monoclonal cells; step four, putting target cells in an operation needle into the culture solution microdrops obtained in the step one in such a manner of carrying one cell in each drop by using the micromanipulation system prepared in the second step, and then carrying out cell culture; step five, replacing the culture solution by a half amount for the cells cultured in the step four two days later after culturing; sixth step six, carrying out enlarge culturing to realize single cell cloning. The method is characterized in that a paraffin oil between the culture solution microdrops isolates the contact of the microdrops with each other, and thus the unicity of cell cloning is guaranteed; meanwhile, as the volume of the culture solution microdrops is less than 5 microliters, biological factors secreted by the cultured cells are not diluted excessively so as to provide a good microenvironment for the growth of cells.

The present invention relates to clonal expansion of somatic cells in subjects, and acquired selective advantage of cell clones during the lifetime of a subject. In particular, the invention relates to methods for predicting the development of cancer based on the observation of specific genetic mutations in somatic cell clones, as well as to methods for treating or preventing cancer in a subject, in which clonal expansion of cells comprising specific modifications is observed.

The invention provides cytokine induced killer (CIK) cell populations and methods of using CIK cells to treat cellular proliferative disorders. CIK cells generated in vitro include both bulk cultures and clones. Individual CIK cell clones display distinct but overlapping lytic specificities for tumor cells and endothelial cells in vitro. When injected in vivo, bulk CIK cell cultures selectively attack tumor tissue. CIK cells can be used to treat a variety of cellular proliferative disorders, including early and late stage cancers as well as hematopoietic cell and solid tissue tumors.

The invention relates to an application of a flavonoid compound, the flavonoid compound is 3, 3', 4', 5, 7-pentahydroxyflavone dihydrate, the ELISA and protein fluorescence quenching experiments provethat the compound can effectively inhibit interaction of PD-1 / PD-L1 proteins, and has a strong affinity for PD-L1 protein. Cell clone number formation and ELISA detection of IL-2 secretion assay prove that the compound can effectively enhance the tumor killing activity of T lymphocyte Jurkat cells against MDA-MB-231 and NCI-H460 tumor cells as well as the expression level of IL-2, and then reverse T cellular immunosuppression. The research results of the application provide a drug for tumor immunotherapy targeting the PD-1 / PD-L1 immunological test site.

The invention provides a method for separating and purifying mesenchymal stem cells originated from formation tissues, the invention implements adherence screening of different time periods in accordance with the adherence ability of different types of the cells in the formation tissues, screens adherent cells having the optimal adherence time period after a specific surface antigen assay of the mesenchymal stem cells of the adherent cell in different time periods, and performs clone culture formed by cell colony subsequent to collecting and suspending the cells so as to obtain the purified mesenchymal stem cells of the formation tissues. The invention has the beneficial effects that: (1) the inventive method can reach relatively good screening effect; (2) using a single-cell clone method for the cells screened by the optimal adherence time period further improves the purification rate of the mesenchymal stem cells of the formation tissues; (3) the mesenchymal stem cells of the formation tissues screened by the optimal adherence time period have high proliferation potential; (4) the mesenchymal stem cells of the formation tissues screened by the optimal adherence time period has outstanding multi-differentiation potential.

The invention discloses a coupling product of streptomycin and carrier protein, and a preparation method of a streptomycin antibody and an application thereof, which relate to a coupling method of the carrier protein such as keyhole limpet hemocyanin, human serum albumin, cow serum albumin, ovalbumin and the like with streptomycin, and immune BALB / C mouse spleen cells are fused with SP2 / 0 mouse myeloma cells by streptomycin immunogens coupled with the carrier protein, and the streptomycin coupled with the carrier protein is used as a coating antigen for screening positive hybridoma and hybridoma that can steady passage and excrete anti-specificity streptomycin monoclonal antibody can be obtained by the cell clone, and ascites monoclonal antibody is prepared. The prepared monoclonal antibody is used for building a direct competition ELISA method with high specificity, sensitivity and accuracy to the streptomycin and immunity colloidal gold test strips. The coupling of the streptomycin and the carrier protein and the preparation method of the streptomycin monoclonal antibody can provide services for detecting streptomycin residue in foods quickly.

The invention relates to chicken EPGCs single cell cloning method. Its feature is that using the fourth generation chicken EPGCs to prepare single cell cloning; culturing and passage to gain heredity uniform chicken EPGCs cell. The invention has simple and practical method, no need special apparatus demand, uses regular cell culture to finish single cell cloning, processes morphology and staining identification for the gained cell to improve that it has its primary characteristic. The produced cell can be directly used in developmental biology study, genome imprinting study fields etc. Because of the cell has little heredity changeability, it can be used to study its different affecting factors.

The invention discloses a screening report vector and screening method for enriching CRISPR / Cas9-mediated homologous recombination repairing cells. The screening report vector includes a universal sgRNA expression cassette, a Cas9 expression cassette, a resistance gene expression cassette and a resistance gene homologous recombination repair template, and the screening reporter vector itself can be repaired by homologous recombination of the resistance gene sequence to make the cells have resistance. The screening report vector, a targeting vector for cell genome HDR editing and a donor vectorform a co-transfection system which can be applied to HDR positive cell cloning with efficient enrichment point editing, fragment insertion and fragment deletion, and can be widely applied to a variety of cells of mammals. The screening efficiency of HDR-edited cells is significantly improved, the reintroduction of double-strand breaking and DNA integration in the unrelated positions of the cellgenome is not required, and an effective way is provided to promote research and application of accurate gene editing.

The invention relates to a preparation method of an inducible pluripotent stem cell of a goat. The preparation method comprises the following steps: A) constructing a lentiviral vector carrying a transcription factor which is selected from Oct4, Sox2, c-Myc, Klf4, Lin28 and Nanog; and B) infecting a goat adult cell by combining the transcription factor with the lentiviral vector prepared in the step A, selecting clone with a shape similar to an embryonic stem cell for subculturing, and preparing the inducible pluripotent stem cell of the goat by screening cell clone in accordance with the characteristics of the embryonic stem cell. According to the preparation method, an optimal culture condition and method of a goat ES (embryonic stem) cell line construction can be established; the inducible pluripotent stem cell of the goat is an excellent vector for goat gene targeting, and the inducible pluripotent stem cell of the goat facilitates the revelation of each gene function and the complex development of the goat.