Journal of Applied Bioanalysis openaccess http://dx.doi.org/10.17145/jab.20.005 REVIEW Engineered Technologies and Bioanalysis of multispecific Antibody Formats Marta Amaral, Soraya Hölper, Christian Lange, Jennifer Jung, Hanno Sjuts, Sandra Weil, Melanie Fischer, Katarina Rado�evi�, Ercole Rao* Sanofi R&D, Biologics Research. *Correspondence: Sanofi-Aventis Deutschland GmbH, R&D, Biologics Research, Industriepark Höchst, H812, 65926 Frankfurt am Main, Germany. Phone: +49 6930515147. Email:
[email protected] Citation: Amaral M, Hölper S, Lange C, Jung J, Sjuts H, Weil S, Fischer M, Rado�evic K, Rao E. Engineered Technologies and Bioanalysis of multispecific ABSTRACT antibody formats. J Appl Bioanal 6(1), The idea of designing multispecific antibodies capable of simultaneously 26-51 (2020). engaging two or more epitopes on the same or different antigens was developed more than 50 years ago. However, the molecular complexity of such Editor: molecules may pose significant challenges for their development and clinical Dr. Lin-zhi Chen, Boehringer use. Particularly challenging is to obtain the correctly assembled combination of Ingelheim Pharmaceuticals Inc., different polypeptide chains, which places significant demand on downstream Ridgefield, CT 06877, USA. process development, analytical characterization and control strategy. Here, we review the progress made in protein engineering to force the correct assembly of Received: July 29, 2019. different heavy and light chains, as well as upstream and downstream processes Revised: September 13, 2019. currently applied to control generation of unwanted byproduct species. We Accepted: September 16, 2019. cover in-depth the analytical methods available to characterize such complex molecules, focusing on mispairing analysis and functional characterization.