8GUZ

Crystal structure of anti-FIXa IgG fab with FAST-Ig mutations

Summary for 8GUZ

• Entry DOI: 10.2210/pdb8guz/pdb
• Descriptor: Anti-factor IXa IgG fab heavy chain, Anti-factor IXa IgG fab light chain, 1,2-ETHANEDIOL, ... (4 entities in total)
• Functional Keywords: bispecific antibody, fast-ig, nxt007, hemophilia a, factor ixa, immune system
• Biological source: Homo sapiens; More
• Total number of polymer chains: 4
• Total formula weight: 97666.42

Authors

Koga, H.,Yamano, T.,Fukami, T.A.,Sampei, Z.,Shiraiwa, H.,Torizawa, T. (deposition date: 2022-09-14, release date: 2023-06-28, Last modification date: 2024-10-23)

Primary citation

Koga, H.,Yamano, T.,Betancur, J.,Nagatomo, S.,Ikeda, Y.,Yamaguchi, K.,Nabuchi, Y.,Sato, K.,Teranishi-Ikawa, Y.,Sato, M.,Hirayama, H.,Hayasaka, A.,Torizawa, T.,Haraya, K.,Sampei, Z.,Shiraiwa, H.,Kitazawa, T.,Igawa, T.,Kuramochi, T.
Efficient production of bispecific antibody by FAST-Ig TM and its application to NXT007 for the treatment of hemophilia A.
Mabs, 15:2222441-2222441, 2023

PubMed Abstract: Efficient production of bispecific antibodies (BsAbs) in single mammalian cells is essential for basic research and industrial manufacturing. However, preventing unwanted pairing of heavy chains (HCs) and light chains (LCs) is a challenging task. To address this, we created an engineering technology for preferential cognate HC/LC and HC/HC paring called FAST-Ig (Four-chain Assembly by electrostatic Steering Technology - Immunoglobulin), and applied it to NXT007, a BsAb for the treatment of hemophilia A. We introduced charged amino-acid substitutions at the HC/LC interface to facilitate the proper assembly for manufacturing a standard IgG-type BsAb. We generated CH1/CL interface-engineered antibody variants that achieved > 95% correct HC/LC pairing efficiency with favorable pharmacological properties and developability. Among these, we selected a design (C3) that allowed us to separate the mis-paired species with an unintended pharmacological profile using ion-exchange chromatography. Crystal structure analysis demonstrated that the C3 design did not affect the overall structure of both Fabs. To determine the final design for HCs-heterodimerization, we compared the stability of charge-based and knobs into hole-based Fc formats in acidic conditions and selected the more stable charge-based format. FAST-Ig was also applicable to stable CHO cell lines for industrial production and demonstrated robust chain pairing with different subclasses of parent BsAbs. Thus, it can be applied to a wide variety of BsAbs both preclinically and clinically.

PubMed: 37339067
DOI: 10.1080/19420862.2023.2222441

Experimental method

X-RAY DIFFRACTION (1.79 Å)