The first trifunctional antibody successfully produced from quadromas is catumaxomab. At first, mouse hybridoma cells whose monoclonal antibodies target one of the desired antigens are produced. Independently, rat hybridoma cells targeting the other antigen are produced. Then, these two cell types are hybridized, yielding hybrid-hybridomas or quadromas, which produce hybrid (trifunctional) antibody as well as pure mouse and pure rat antibody. The trifunctional antibody is extracted chromatographically with protein A.
The disadvantage of quadromas is the produced antibodies are a mixture. Especially if these two hybridoma cells are the same species, up to ten different kinds of antibody can be produced, most of which have mismatched heavy or light chains, or both. Fig. 1 below presents the possible combinations of light- and heavy-chains in antibodies produced by quadroma cell lines.
Fig. 1 Antibodies produced by quadroma cell lines. The seven antibodies on the left have at least one mismatched binding region. Of the three antibodies on the right, two are not hybridized, and the remaining (rightmost) is the desired trifunctional antibody.
Scientists in Creative Biolabs have successfully established the method to produce custom bispecific antibody according to client's requirement. If interested, please contact us.
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