The bispecific antibody therapeutic field is rapidly growing. Since 2011, over 400 bispecific molecules have successfully entered clinical trials; and 2023 alone saw an introduction of 120 new molecules.
However, as Jin Lu, Senior Technical Support Manager at Lonza, points out in her talk, the production of bispecifics can still be challenging. In her talk, Lu walks us through how knob-in-hole technology was able to cement itself as the gold standard solution for heavy chain/heavy chain mispairing.
“However, even if you have correctly assembled heavy chain/heavy chain product, light chain/heavy chain mispairing can still occur. At Lonza, we have developed bYlok to tackle this particular challenge,” says Lu.
Lonza’s bYlok uses antibody engineering to eliminate native disulfide bonds and create artificial ones, improving heavy-light chain pairing.
Using a series of experimental case studies, Lu demonstrates how bYlok technology, combined with knob-into-hole, can significantly increase the yield of correctly assembled bispecific antibodies. Furthermore, characterization of bYlok bispecifics reveal how the platform is able to retain antigen binding properties, comparable affinity, and binding kinetics to controls, with no increase in immunogenicity.
Lu proves the efficacy of bYlok by comparing the platform to other solutions: DM-1 and DM-2, demonstrating more consistent product assembly and higher titers.
“The bYlok technology provides an easy to implement solution for heavy chain and light chain mispairing and for the manufacturing of IgG like bispecific antibodies.”
Lu continues: “It has minimal modification to the monoclonal antibody design. It fits into a standard manufacturing workflow. It generates more than 95% correct heavy/light chain pairing. It's a portable and versatile platform. And the data I’ve demonstrated today shows that the immunogenicity, thermal stability, and binding properties of the bispecific antibodies are not affected.”
