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Bispecific antibodies (BisAbs) represent a major breakthrough in biotherapeutics by combining the functionalities of two different antibodies to unlock their potential for a wide range of applications. Conventional BisAb production has relied on labor-intensive and complex genetic engineering methods. This chapter introduces a cutting-edge protocol that circumvents the need for genetic modifications by using chemical conjugation strategies. We present a streamlined, fully chemical methodology for generating BisAbs that leverages the precision of the AJICAP second-generation technology for accurate site-specific conjugation. In addition, this chapter explores the application of haloketone chemistry as an improved and more effective replacement for traditional maleimide-based crosslinking techniques for the fusion of two antibody-related proteins. The analytical performance and results of the newly developed BisAb were thoroughly investigated, highlighting its high efficiency and effectiveness. This innovative approach represents a significant step forward in the production of BisAbs and offers a simpler, more accessible, and potentially scalable alternative to traditional genetic engineering techniques.

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Enhanced Chemical Method for Bispecific Antibody Production: Leveraging Site-Specific Conjugation and Stable Crosslinker Technology

  • 实验方案

  • 发表日期：

    2026年

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  • Tomohiro Fujii,
  • Yutaka Matsuda

摘要

Bispecific antibodies (BisAbs) represent a major breakthrough in biotherapeutics by combining the functionalities of two different antibodies to unlock their potential for a wide range of applications. Conventional BisAb production has relied on labor-intensive and complex genetic engineering methods. This chapter introduces a cutting-edge protocol that circumvents the need for genetic modifications by using chemical conjugation strategies. We present a streamlined, fully chemical methodology for generating BisAbs that leverages the precision of the AJICAP second-generation technology for accurate site-specific conjugation. In addition, this chapter explores the application of haloketone chemistry as an improved and more effective replacement for traditional maleimide-based crosslinking techniques for the fusion of two antibody-related proteins. The analytical performance and results of the newly developed BisAb were thoroughly investigated, highlighting its high efficiency and effectiveness. This innovative approach represents a significant step forward in the production of BisAbs and offers a simpler, more accessible, and potentially scalable alternative to traditional genetic engineering techniques.