A novel efficient bispecific antibody format, combining a conventional antigen-binding fragment with a single domain antibody, avoids potential heavy-light chain mis-pairing

Shuyu Huang, Aina Segués, David Lutje Hulsik, Dietmar M. Zaiss, Alice J.a.m. Sijts, Sander M.j. Van Duijnhoven, Andrea Van Elsas

Research output: Contribution to journalArticlepeer-review

Abstract

Due to the technical innovations in generating bispecific antibodies (BsAbs) in recent years, BsAbs have become important reagents for diagnostic and therapeutic applications. However, the difficulty of producing a heterodimer consisting of two different arms with high yield and purity constituted a major limitation for their application in academic and clinical settings. Here, we describe a novel Fc-containing BsAb format (Fab × sdAb-Fc) composed of a conventional antigen-binding fragment (Fab), and a single domain antibody (sdAb), which avoids heavy-light chain mis-pairing during antibody assembly. In this study, the Fab x sdAb-Fc BsAbs were efficiently produced by three widely used heavy-heavy chain heterodimerization methods: Knobs-into-holes (KIH), Charge-pairs (CP) and controlled Fab-arm exchange (cFAE), respectively. The novel Fab x sdAb-Fc format provided a rapid and efficient strategy to generate BsAb with high purity and a unique possibility to further purify desired BsAbs from undesired antibodies based on molecular weight (MW). Compared to conventional BsAb formats, the advantages of Fab x sdAb-Fc format may thus provide a straightforward opportunity to apply bispecific antibody principles to research and development of novel targets and pathways in diseases such as cancer and autoimmunity.
Original languageEnglish
Article number112811
JournalJournal of Immunological Methods
Volume483
Early online date19 Jun 2020
DOIs
Publication statusE-pub ahead of print - 19 Jun 2020

Keywords

  • bispecific antibody
  • antibody chain association
  • knobs into holes
  • charge pairs
  • controlled fab arm exchange

Fingerprint Dive into the research topics of 'A novel efficient bispecific antibody format, combining a conventional antigen-binding fragment with a single domain antibody, avoids potential heavy-light chain mis-pairing'. Together they form a unique fingerprint.

Cite this