2013/03/11 by Aran F. Labrijn, Joyce Meesters, Bart E.C.G. de Goeij +11 · 1 voice · 1 citation
Biochemistry, Genetics and Molecular Biology · Medicine · #Glycosylation and Glycoproteins Research #Monoclonal and Polyclonal Antibodies Research #Protein purification and stability
paper · pdf · doi:10.1073/pnas.1220145110
openalex publication_date 2013/03/11 · openalex created_date 2025/10/10 · openalex updated_date 2026/07/15
The promise of bispecific antibodies (bsAbs) to yield more effective therapeutics is well recognized; however, the generation of bsAbs in a practical and cost-effective manner has been a formidable challenge. Here we present a technology for the efficient generation of bsAbs with normal IgG structures that is amenable to both antibody drug discovery and development. The process involves separate expression of two parental antibodies, each containing single matched point mutations in the CH3 domains. The parental antibodies are mixed and subjected to controlled reducing conditions in vitro that separate the antibodies into HL half-molecules and allow reassembly and reoxidation to form highly pure bsAbs. The technology is compatible with standard large-scale antibody manufacturing and ensures bsAbs with Fc-mediated effector functions and in vivo stability typical of IgG1 antibodies. Proof-of-concept studies with HER2×CD3 (T-cell recruitment) and HER2×HER2 (dual epitope targeting) bsAbs demonstrate superior in vivo activity compared with parental antibody pairs.