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

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Kummer, J. Alain and Kamp, Angela M. and Katwijk, Marcel van and Brakenhoff, Just P.J. and Radosevic, Katarina and Leeuwen, Anne Marie van and Borst, Jannie and Verweij, Cornelis L. and Hack, C. Erik (1993) Production and characterization of monoclonal antibodies raised against recombinant human granzymes A and B and showing cross reactions with the natural proteins. Journal of Immunological Methods, 163 (1). pp. 77-83. ISSN 0022-1759

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Abstract: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.
Item Type:Article
Copyright:© 1993 Elsevier Science
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Link to this item:http://purl.utwente.nl/publications/24593
Official URL:http://dx.doi.org/10.1016/0022-1759(93)90241-X
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Metis ID: 129394