TY - JOUR T1 - Generation of dual specific bivalent BiTEs (dbBIspecific T-cell Engaging antibodies) for cellular immunotherapy JF - bioRxiv DO - 10.1101/559427 SP - 559427 AU - Maciej Kujawski AU - Lin Li AU - Supriyo Bhattacharya AU - Patty Wong AU - Wen-Hui Lee AU - Lindsay Williams AU - Harry Li AU - Junie Chea AU - Kofi Poku AU - Nicole Bowles AU - Nagarajan Vaidehi AU - Paul Yazaki AU - John E. Shively Y1 - 2019/01/01 UR - http://biorxiv.org/content/early/2019/02/24/559427.abstract N2 - Bispecific T-cell engaging antibodies (BiTES), comprising dual anti-CD3 and anti-tumor antigen scFv fragments, are important therapeutic agents for the treatment of cancer. The dual scFv construct for BiTES requires proper protein folding while their small molecular size leads to rapid kidney clearance. Here we show that an intact (150 kDa) anti-tumor antigen antibody to CEA was joined in high yield (ca. 30%) to intact (150 kDa) anti-murine and anti-human CD3 antibodies using hinge region specific Click chemistry to form dual-specific, bivalent BiTES (db BiTES, 300 kDa). The interlocked hinge regions are compatible with a structural model that fits the electron micrographs of the 300 kDa particles. Compared to intact anti-CEA antibody, dbBiTES maintain high in vivo tumor targeting as demonstrated by PET imaging, and redirect dbBiTE coated T-cells (1 microgram/10 million cells) to kill CEA+ target cells both in vitro, and in vivo in CEA transgenic mice. ER -