6OSN

Potent and Selective Antitumor Antibody Targeting a Membrane-Proximal Epitope of ROR2

Summary for 6OSN

• Entry DOI: 10.2210/pdb6osn/pdb
• Descriptor: Tyrosine-protein kinase transmembrane receptor ROR2, ACETATE ION (3 entities in total)
• Functional Keywords: single chain fv, scfv, antibody, ror2, kringle domain, receptor tyrosine kinase-like orphan receptor, phage display, immune system
• Biological source: Homo sapiens (Human)
• Total number of polymer chains: 1
• Total formula weight: 11017.22

Authors

Park, H.,Rader, C. (deposition date: 2019-05-01, release date: 2020-04-01, Last modification date: 2024-10-16)

Primary citation

Goydel, R.S.,Weber, J.,Peng, H.,Qi, J.,Soden, J.,Freeth, J.,Park, H.,Rader, C.
Affinity maturation, humanization, and co-crystallization of a rabbit anti-human ROR2 monoclonal antibody for therapeutic applications.
J.Biol.Chem., 295:5995-6006, 2020

PubMed Abstract: Antibodies are widely used as cancer therapeutics, but their current use is limited by the low number of antigens restricted to cancer cells. A receptor tyrosine kinase, receptor tyrosine kinase-like orphan receptor 2 (ROR2), is normally expressed only during embryogenesis and is tightly down-regulated in postnatal healthy tissues. However, it is up-regulated in a diverse set of hematologic and solid malignancies, thus ROR2 represents a candidate antigen for antibody-based cancer therapy. Here we describe the affinity maturation and humanization of a rabbit mAb that binds human and mouse ROR2 but not human ROR1 or other human cell-surface antigens. Co-crystallization of the parental rabbit mAb in complex with the human ROR2 kringle domain (hROR2-Kr) guided affinity maturation by heavy-chain complementarity-determining region 3 (HCDR3)-focused mutagenesis and selection. The affinity-matured rabbit mAb was then humanized by complementarity-determining region (CDR) grafting and framework fine tuning and again co-crystallized with hROR2-Kr. We show that the affinity-matured and humanized mAb retains strong affinity and specificity to ROR2 and, following conversion to a T cell-engaging bispecific antibody, has potent cytotoxicity toward ROR2-expressing cells. We anticipate that this humanized affinity-matured mAb will find application for antibody-based cancer therapy of ROR2-expressing neoplasms.

PubMed: 32193207
DOI: 10.1074/jbc.RA120.012791

Experimental method

X-RAY DIFFRACTION (1.083 Å)