Antibody mimetic drug conjugate manufactured by high-yield Escherichia coli expression and non-covalent binding system.

Abstract

Antibody-drug conjugates (ADCs) are a major therapeutic tool for the treatment of advanced cancer. Malignant cells in advanced cancer often display multiple genetic mutations and become resistant to monotherapy. Therefore, a therapeutic regimen that simultaneously targets multiple molecules with multiple payloads is desirable. However, the development of ADCs is hampered by issues in biopharmaceutical manufacturing and the complexity of the conjugation process of low-molecular-weight payloads to biologicals. Here, we report antibody mimetic-drug conjugates (AMDCs) developed by exploiting the non-covalent binding property of payloads based on high-affinity binding of mutated streptavidin and modified iminobiotin. Miniprotein antibodies were fused to a low immunogenic streptavidin variant, which was then expressed in Escherichia coli inclusion bodies, solubilized, and refolded into functional tetramers. The AMDC developed against human epidermal growth factor receptor 2 (HER2) effectively killed cultured cancer cells using bis-iminobiotin conjugated to photo-activating silicon phthalocyanine. The HER2-targeting AMDC was also effective in vivo against a mouse KPL-4 xenograft model. This AMDC platform provides rapid, stable, and high-yield therapeutics against multiple targets.

• Authors: Kenzo Yamatsugu, Hiroto Katoh, Takefumi Yamashita, Kazuki Takahashi, Sho Aki, Toshifumi Tatsumi, Yudai Kaneko, Takeshi Kawamura, Mai Miura, Masazumi Ishii, Kei Ohkubo, Tsuyoshi Osawa, Tatsuhiko Kodama, Shumpei Ishikawa, Motomu Kanai, Akira Sugiyama
• Journal: Protein expression and purification (Protein Expr Purif) Vol. 192 Pg. 106043 (04 2022) ISSN: 1096-0279 [Electronic] United States
• PMID: 34973460 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2021 Elsevier Inc. All rights reserved.

Chemical References

• Immunoconjugates
• iminobiotin
• Biotin
• Streptavidin
• ERBB2 protein, human
• Receptor, ErbB-2

Topics

• Animals
• Biotin (administration & dosage, analogs & derivatives, chemistry, genetics, immunology)
• Cell Line, Tumor
• Cloning, Molecular
• Escherichia coli (genetics, metabolism)
• Gene Expression
• Humans
• Immunoconjugates (administration & dosage, chemistry, genetics, immunology)
• Mice
• Mice, Inbred BALB C
• Neoplasms (drug therapy)
• Protein Folding
• Receptor, ErbB-2 (antagonists & inhibitors, genetics, immunology)
• Streptavidin (administration & dosage, chemistry, genetics, immunology)