Production and characterization of monoclonal antibodies raised against recombinant human granzymes A and B and showing cross reactions with the natural proteins

J. Alain Kummer, Angela M. Kamp, Marcel van Katwijk, Just P.J. Brakenhoff, Katarina Radosevic, K. Radosevic, Anne Marie van Leeuwen, Jannie Borst, Cornelis L. Verweij, C. Erik Hack

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    The human serine proteases granzymes A and B are expressed in cytotoplasmic granules of activated cytotoxic T lymphocytes and natural killer cells. Recombinant granzyme A and granzyme B proteins were produced in bacteria, purified and then used to raise specific mouse monoclonal antibodies. Seven monoclonal antibodies (mAb) were raised against granzyme A, which all recognized the same or overlapping epitopes. They reacted specifically in an immunoblot of interleukin-2 (IL-2) stimulated PBMNC with a disulfide-linked homodimer of 43 kDa consisting of 28 kDa subunits. Seven mAb against granzyme B were obtained, which could be divided into two groups, each recognizing a different epitope. On an immunoblot, all mAb reacted with a monomer of 33 kDa protein. By immunohistochemistry, these mAb could be used to detect granzymes A and B expression in activated CTL and NK cells. The availability of these mAb may facilitate studies on the role of human cytotoxic cells in various immune reactions and may contribute to a better understanding of the role of granzmes A and B in the cytotoxic response in vivo.
    Original languageUndefined
    Pages (from-to)77-83
    Number of pages7
    JournalJournal of immunological methods
    Issue number1
    Publication statusPublished - 1993


    • Granzyme B
    • Granzyme A
    • METIS-129394
    • Monoclonal antibody
    • Immunohistochemical staining
    • IR-24593

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