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Antibody Production

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Mark Brunswick – One of the best experts on this subject based on the ideXlab platform.

  • In vitro Antibody Production.
    Current protocols in molecular biology edited by Frederick M. Ausubel … [et al.], 2001
    Co-Authors: James J. Mond, Mark Brunswick

    Abstract:

    This unit describes the antigenic stimulation of in vitro Antibody Production by B cells and the subsequent measurement of secreted antibodies. A generalized system for inducing in vitro Antibody Production is presented along with a procedure for quantifying the number of Antibody-producing cells by plaque-forming cell (PFC) assays: the Cunningham-Szenberg technique and the Jerne-Nordin technique. The assay can be modified as described to measure all classes of antibodies or to enumerate total immunoglobulin-secreting B cells. A protocol for preparing the resting B cells by Percoll gradient centrifugation is also described.

  • Current Protocols in Molecular Biology – In Vitro Antibody Production
    Current Protocols in Molecular Biology, 2000
    Co-Authors: James J. Mond, Mark Brunswick

    Abstract:

    This unit describes the antigenic stimulation of in vitro Antibody Production by B cells and the subsequent measurement of secreted antibodies. A generalized system for inducing in vitro Antibody Production is presented along with a procedure for quantifying the number of Antibody-producing cells by plaque-forming cell (PFC) assays: the Cunningham-Szenberg technique and the Jerne-Nordin technique. The assay can be modified as described to measure all classes of antibodies or to enumerate total immunoglobulin-secreting B cells. A protocol for preparing the resting B cells by Percoll gradient centrifugation is also described.

James J. Mond – One of the best experts on this subject based on the ideXlab platform.

  • In vitro Antibody Production.
    Current protocols in molecular biology edited by Frederick M. Ausubel … [et al.], 2001
    Co-Authors: James J. Mond, Mark Brunswick

    Abstract:

    This unit describes the antigenic stimulation of in vitro Antibody Production by B cells and the subsequent measurement of secreted antibodies. A generalized system for inducing in vitro Antibody Production is presented along with a procedure for quantifying the number of Antibody-producing cells by plaque-forming cell (PFC) assays: the Cunningham-Szenberg technique and the Jerne-Nordin technique. The assay can be modified as described to measure all classes of antibodies or to enumerate total immunoglobulin-secreting B cells. A protocol for preparing the resting B cells by Percoll gradient centrifugation is also described.

  • Current Protocols in Molecular Biology – In Vitro Antibody Production
    Current Protocols in Molecular Biology, 2000
    Co-Authors: James J. Mond, Mark Brunswick

    Abstract:

    This unit describes the antigenic stimulation of in vitro Antibody Production by B cells and the subsequent measurement of secreted antibodies. A generalized system for inducing in vitro Antibody Production is presented along with a procedure for quantifying the number of Antibody-producing cells by plaque-forming cell (PFC) assays: the Cunningham-Szenberg technique and the Jerne-Nordin technique. The assay can be modified as described to measure all classes of antibodies or to enumerate total immunoglobulin-secreting B cells. A protocol for preparing the resting B cells by Percoll gradient centrifugation is also described.

Daisuke Nishimiya – One of the best experts on this subject based on the ideXlab platform.

  • Proteins improving recombinant Antibody Production in mammalian cells
    Applied Microbiology and Biotechnology, 2014
    Co-Authors: Daisuke Nishimiya

    Abstract:

    Mammalian cells have been successfully used for the industrial manufacture of antibodies due to their ability to synthesize antibodies correctly. Nascent polypeptides must be subjected to protein folding and assembly in the ER and the Golgi to be secreted as mature proteins. If these reactions do not proceed appropriately, unfolded or misfolded proteins are degraded by the ER-associated degradation (ERAD) pathway. The accumulation of unfolded proteins or intracellular Antibody crystals accompanied by this failure triggers the unfolded protein response (UPR), which can considerably attenuate the levels of translation, folding, assembly, and secretion, resulting in reduction of Antibody productivity. Accumulating studies by omics-based analysis of recombinant mammalian cells suggest that not only protein secretion processes including protein folding and assembly but also translation are likely to be the rate-limiting factors for increasing Antibody Production. Here, this review describes the mechanism of Antibody folding and assembly and recent advantages which could improve recombinant Antibody Production in mammalian cells by utilizing proteins such as ER chaperones or UPR-related proteins.