Ten years in the making: application of CrossMab technology for the development of therapeutic bispecific antibodies and antibody fusion proteins

Abstract

Bispecific antibodies have recently attracted intense interest. CrossMab technology was described in 2011 as novel approach enabling correct antibody light-chain association with their respective heavy chain in bispecific antibodies, together with methods enabling correct heavy-chain association using existing pairs of antibodies. Since the original description, CrossMab technology has evolved in the past decade into one of the most mature, versatile, and broadly applied technologies in the field, and nearly 20 bispecific antibodies based on CrossMab technology developed by Roche and others have entered clinical trials. The most advanced of these are the Ang-2/VEGF bispecific antibody faricimab, currently undergoing regulatory review, and the CD20/CD3 T cell bispecific antibody glofitamab, currently in pivotal Phase 3 trials. In this review, we introduce the principles of CrossMab technology, including its application for the generation of bi-/multispecific antibodies with different geometries and mechanisms of action, and provide an overview of CrossMab-based therapeutics in clinical trials.

Aflibercept versus Faricimab in the Treatment of Neovascular Age-Related Macular Degeneration and Diabetic Macular Edema: A Review

“…Faricimab, known at the stage of preclinical experiments as RG7716, is the first bispecific monoclonal antibody designed for intravitreal use and was approved for the treatment of nAMD and DME by the FDA in January 2021 ( Table 1 ) [ 33 ]. Bispecific heterodimeric antibodies, due to having different light chains in each of the fragment antigen-binding (Fab) regions, are characterized by the possibility of binding two different targets, which is unattainable for traditional monospecific antibodies [ 69 ]. These properties were achieved through a knob and hole mechanism between the heavy chains during the drug design process [ 8 ] with the CrossMAb CH1-CL technology, first described in 2011 ( Figure 1 ) [ 69 ].…”

“…Bispecific heterodimeric antibodies, due to having different light chains in each of the fragment antigen-binding (Fab) regions, are characterized by the possibility of binding two different targets, which is unattainable for traditional monospecific antibodies [ 69 ]. These properties were achieved through a knob and hole mechanism between the heavy chains during the drug design process [ 8 ] with the CrossMAb CH1-CL technology, first described in 2011 ( Figure 1 ) [ 69 ]. Due to its specific structure, the faricimab molecule has one VEGF-binding domain and one Ang-2-binding domain [ 8 ], which enables their simultaneous and independent neutralization [ 33 , 70 ].…”

Antibodies to watch in 2022

“…Glofitamab (RO7082859, CD20-TCB, RG6026) is a full-length IgG1λ/ҡ, asymmetric 2:1 CrossMab. 153 Like mosunetuzumab, which is also being developed by Roche, glofitamab is a bispecific T-cell engaging antibody that targets CD20 and CD3. The two molecules, however, are structurally distinct.…”

Bispecific Antibody-Based Immune-Cell Engagers and Their Emerging Therapeutic Targets in Cancer Immunotherapy

“…CrossMab technology is developed to inhibit the mispairing of light chains [ 251 ]. However, it takes a different approach from that of the common light-chain technology.…”

“…A phase III clinical study (NCT05083169) aimed at evaluating the efficacy of teclistamab in combination with daratumumab, which is a US FDA-approved anti-CD38 mAb, in the treatment of patients with R/R MM is ongoing [ 284 ]. Other CD31- and BCMA-specific bsAb-based T-cell engagers, such as elranatamab (PF-06863135), linvoseltamab (REGN5458), REGN5459, EMB-06, CC-93269 (EM801), TNB-383B, and AMG 701, are being actively assessed in various clinical studies [ 251 , 285 ].…”