The idea of designing multispecific antibodies capable of simultaneously engaging two or more epitopes on the same or different antigens was developed more than 50 years ago. However, the molecular complexity of such molecules may pose significant challenges for their development and clinical use. Particularly challenging is to obtain the correctly assembled combination of different polypeptide chains, which places significant demand on downstream process development, analytical characterization and control strategy. Here, we review the progress made in protein engineering to force the correct assembly of different heavy and light chains, as well as upstream and downstream processes currently applied to control generation of unwanted byproduct species.
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