Facile generation of biepitopic antibodies with intrinsic agonism for activating tumor necrosis factor receptors

Harkamal S Jhajj; John S Schardt; Namir Khalasawi; Emily L Yao; Timon S Lwo; Na-Young Kwon; Ryen L O'Meara; Alec A Desai; Peter M Tessier

doi:10.1016/j.chembiol.2024.03.010

Facile generation of biepitopic antibodies with intrinsic agonism for activating tumor necrosis factor receptors

Cell Chem Biol. 2024 May 16;31(5):944-954.e5. doi: 10.1016/j.chembiol.2024.03.010. Epub 2024 Apr 22.

Authors

Harkamal S Jhajj¹, John S Schardt², Namir Khalasawi³, Emily L Yao⁴, Timon S Lwo¹, Na-Young Kwon², Ryen L O'Meara⁵, Alec A Desai⁵, Peter M Tessier⁶

Affiliations

¹ Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI 48109, USA; Biointerfaces Institute, University of Michigan, Ann Arbor, MI 48109, USA.
² Department of Pharmaceutical Sciences, University of Michigan, Ann Arbor, MI 48109, USA; Department of Chemical Engineering, University of Michigan, Ann Arbor, MI 48109, USA; Biointerfaces Institute, University of Michigan, Ann Arbor, MI 48109, USA.
³ Department of Pharmaceutical Sciences, University of Michigan, Ann Arbor, MI 48109, USA; Biointerfaces Institute, University of Michigan, Ann Arbor, MI 48109, USA.
⁴ Department of Chemistry, University of Michigan, Ann Arbor, MI 48109, USA; Biointerfaces Institute, University of Michigan, Ann Arbor, MI 48109, USA.
⁵ Department of Chemical Engineering, University of Michigan, Ann Arbor, MI 48109, USA; Biointerfaces Institute, University of Michigan, Ann Arbor, MI 48109, USA.
⁶ Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI 48109, USA; Department of Pharmaceutical Sciences, University of Michigan, Ann Arbor, MI 48109, USA; Department of Chemical Engineering, University of Michigan, Ann Arbor, MI 48109, USA; Biointerfaces Institute, University of Michigan, Ann Arbor, MI 48109, USA. Electronic address: ptessier@umich.edu.

PMID: 38653243
DOI: 10.1016/j.chembiol.2024.03.010

Abstract

Agonist antibodies are being pursued for therapeutic applications ranging from neurodegenerative diseases to cancer. For the tumor necrosis factor (TNF) receptor superfamily, higher-order clustering of three or more receptors is key to their activation, which can be achieved using antibodies that recognize two unique epitopes. However, the generation of biepitopic (i.e., biparatopic) antibodies typically requires animal immunization and is laborious and unpredictable. Here, we report a simple method for identifying biepitopic antibodies that potently activate TNF receptors without the need for additional animal immunization. Our approach uses existing, receptor-specific IgGs, which lack intrinsic agonist activity, to block their corresponding epitopes, then selects single-chain antibodies that bind accessible epitopes. The selected antibodies are fused to the light chains of IgGs to generate human tetravalent antibodies. We highlight the broad utility of this approach by converting several clinical-stage antibodies against OX40 and CD137 (4-1BB) into biepitopic antibodies with potent agonist activity.

Keywords: 4-1BB; CD137; FcγR; OX40; TNF; TNFR; agonist; antibody; biparatopic; mAb; receptor clustering.

MeSH terms

Animals
Antibodies / immunology
Epitopes* / chemistry
Epitopes* / immunology
Humans
Mice
Receptors, OX40 / agonists
Receptors, OX40 / antagonists & inhibitors
Receptors, OX40 / immunology
Receptors, OX40 / metabolism
Receptors, Tumor Necrosis Factor / agonists
Receptors, Tumor Necrosis Factor / immunology
Receptors, Tumor Necrosis Factor / metabolism
Single-Chain Antibodies / chemistry
Single-Chain Antibodies / immunology
Single-Chain Antibodies / pharmacology
Tumor Necrosis Factor Receptor Superfamily, Member 9 / agonists
Tumor Necrosis Factor Receptor Superfamily, Member 9 / antagonists & inhibitors
Tumor Necrosis Factor Receptor Superfamily, Member 9 / immunology
Tumor Necrosis Factor Receptor Superfamily, Member 9 / metabolism

Grants and funding

R01 AG080016/AG/NIA NIH HHS/United States