Production of monoclonal camelid antibodies by means of hybridoma technology (P3376)
Pamela Holzloehner, Erik Schliebs, Natalia Maier, Jonas Füner, Burkhard Micheel, Katja Heilmann
Abstract
Pamela Holzloehner, Erik Schliebs, Natalia Maier, Jonas Füner, Burkhard Micheel, Katja Heilmann
Abstract
Abstract Here we describe the first monoclonal immunoglobulins produced by camelid hybridomas. Hybridomas were generated by using the CBF7 heteromyeloma line to fuse peripheral blood lymphocytes from New World camels (llamas, alpacas) and Old World camels (Bactrian camels). The hybridomas produced immunoglobulins of IgG1, IgG2 and IgG3 subclasses as defined by both enzyme immunoassays and PCR and subsequent gene sequencing. The subclass-specific murine monoclonal antibodies were also produced in the experiments described here. They could be used to detect conventional four-chain camelid antibodies (of subclass IgG1) and heavy chain-only antibodies (both of subclasses IgG2 and IgG3). The different monoclonal heavy chain-only immunoglobulins and the producing hybridomas are of special interest since they can be used to produce further secondary reagents (murine monoclonal antibodies) for the characterization of different camelid antibodies and to solve some as yet unresolved problems in the molecular biology of heavy chain antibody-producing cells. Most importantly, the technique described here can be the basis for the production of full-length heavy chain-only monoclonal antibodies of camelids directed against specific antigens.
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Abstract Here we describe the first monoclonal immunoglobulins produced by camelid hybridomas. Hybridomas were generated by using the CBF7 heteromyeloma line to fuse peripheral blood lymphocytes from New World camels (llamas, alpacas) and Old World camels (Bactrian camels). The hybridomas produced immunoglobulins of IgG1, IgG2 and IgG3 subclasses as defined by both enzyme immunoassays and PCR and subsequent gene sequencing. The subclass-specific murine monoclonal antibodies were also produced in the experiments described here. They could be used to detect conventional four-chain camelid antibodies (of subclass IgG1) and heavy chain-only antibodies (both of subclasses IgG2 and IgG3). The different monoclonal heavy chain-only immunoglobulins and the producing hybridomas are of special interest since they can be used to produce further secondary reagents (murine monoclonal antibodies) for the characterization of different camelid antibodies and to solve some as yet unresolved problems in the molecular biology of heavy chain antibody-producing cells. Most importantly, the technique described here can be the basis for the production of full-length heavy chain-only monoclonal antibodies of camelids directed against specific antigens.
Key concepts: Monoclonal antibody, Subclass, Antibody, Biology, Molecular biology, Immunoglobulin light chain, Monoclonal, Antigen