Asymmetric Fc Engineering for Bispecific Antibodies with Reduced Effector Function

Antibodies

Joseph Schrag¹  Yves Durocher¹  Jason Baardsnes¹  Eric Escobar-Cabrera²  Surjit Dixit²  Paula Lario² 

[1] Human Health Therapeutics, National Research Council Canada, 6100 Royalmount Ave, Montreal, QC H4P 2R2, Canada;Zymeworks Inc., 540-1385 West 8th Ave, Vancouver, BC V6H 3V9, Canada;

关键词: Fc engineering;    asymmetric bispecific antibody;    ion exchange chromatography;    knock-out effector function;   

DOI  :  10.3390/antib6020007

来源: DOAJ

【 摘 要 】

Asymmetric bispecific antibodies are a rapidly expanding therapeutic antibody class, designed to recognize two different target epitopes concurrently to achieve novel functions not available with normal antibodies. Many therapeutic designs require antibodies with reduced or silenced effector function. Although many solutions have been described in the literature to knockout effector function, to date all of them have involved the use of a specific antibody subtype (e.g., IgG2 or IgG4), or symmetric mutations in the lower hinge or CH2 domain of traditional homodimeric monospecific antibodies. In the context of a heterodimeric Fc, we describe novel asymmetric Fc mutations with reduced or silenced effector function in this article. These heteromultimeric designs contain asymmetric charged mutations in the lower hinge and the CH2 domain of the Fc. Surface plasmon resonance showed that the designed mutations display much reduced binding to all of the Fc gamma receptors and C1q. Ex vivo ADCC and CDC assays showed a consistent reduction in activity. Differential scanning calorimetry showed increased thermal stability for some of the designs. Finally, the asymmetric nature of the introduced charged mutations allowed for separation of homodimeric impurities by ion exchange chromatography, providing, as an added benefit, a purification strategy for the production of bispecific antibodies with reduced or silenced effector function.

【 授权许可】

Unknown