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FIGURE 1. Schematic representation of how a tri-Fab (TF) bispecific antibody is formed by the DNL method. Component A links a cysteine-modified DDD to a Fab of an antitumor antibody, which results in spontaneous formation of component A2. Component B links an anchoring domain (AD) to a Fab of an antihapten antibody. The AD is modified with cysteine on each end. AD will naturally "dock" with the DDD when components A and B are mixed, which brings the 2 molecules together in a well-defined orientation and also results in disulfide bonds across the 2 proteins.
