21TW
Cryo-EM structure of TNF-alpha in complex with two anti-TNF-alpha nanobodies, TNF30, derived from the TNF-alpha inhibitor Ozoralizumab (OZR)
Summary for 21TW
| Entry DOI | 10.2210/pdb21tw/pdb |
| EMDB information | 67994 |
| Descriptor | anti-TNF-alpha nanobodies, TNF30, derived from Ozoralizumab (OZR), Tumor necrosis factor (2 entities in total) |
| Functional Keywords | nanobody, vhh, tnf-alpha, hsa, ozoralizumab, ozr, cytokine |
| Biological source | Homo sapiens More |
| Total number of polymer chains | 4 |
| Total formula weight | 90754.79 |
| Authors | Tanaka, Y.,Sato, K.,Mima, M.,Mishima-Tsumagari, C. (deposition date: 2025-12-24, release date: 2026-04-29) |
| Primary citation | Mima, M.,Sato, K.,Yokoyama, T.,Mishima-Tsumagari, C.,Ohnuki, T.,Tanaka, Y.,Iwamoto, K. Cryo-EM elucidates the interaction mechanism of ozoralizumab, a humanized anti-TNF alpha NANOBODY® compound. Biochem.Biophys.Res.Commun., 816:153572-153572, 2026 Cited by PubMed Abstract: Ozoralizumab (OZR) is a next-generation TNF inhibitor composed of two identical humanized anti-TNFα NANOBODY® molecules (TNF30s) recombinantly linked via one humanized anti-human serum albumin (HSA) NANOBODY® molecule (ALB8) and two peptide linkers. OZR is designed as a unique format to exert potent inhibitory effects against TNFα with long plasma half-life. However, the three-dimensional structure of OZR-TNFα-HSA complex has not yet been elucidated, and a complete understanding of its interaction mechanism with TNFα is yet to be gained. In this study, we successfully observed the formation of the OZR-TNFα-HSA ternary complex by single-particle cryo-electron microscopy. The single-particle analysis revealed that the two TNF30 molecules of OZR simultaneously bind bivalently to TNFα in a 1:1-bivalent binding mode, while the ALB8 molecule binds to HSA, forming a ternary complex. Thus, OZR exhibits a binding mode significantly different from that of other IgG-type TNFα inhibitors. Furthermore, surface plasmon resonance (SPR) analysis demonstrated that the 1:1-bivalent binding mode confers an exceptionally slow dissociation rate, thereby contributing to the potent TNFα-neutralizing activity of OZR. These findings not only lend support to the favorable clinical efficacy of OZR from a structural standpoint but also lay the foundation for the rational design and development of next-generation TNFα inhibitors with enhanced and sustained efficacy. PubMed: 41935434DOI: 10.1016/j.bbrc.2026.153572 PDB entries with the same primary citation |
| Experimental method | ELECTRON MICROSCOPY (2.43 Å) |
Structure validation
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