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. 2023 Mar 17;18(3):443-448.
doi: 10.1021/acschembio.2c00634. Epub 2023 Mar 8.

Genetic Code Expansion for Site-Specific Labeling of Antibodies with Radioisotopes

Christine Koehler  1   2 , Paul F Sauter  1   2 , Benedikt Klasen  3 , Christopher Waldmann  4 , Stefanie Pektor  4 , Nicole Bausbacher  4 , Edward A Lemke  5   6 , Matthias Miederer  4
Affiliations

Genetic Code Expansion for Site-Specific Labeling of Antibodies with Radioisotopes

Christine Koehler et al. ACS Chem Biol. .

Abstract

Due to their target specificity, antibody-drug conjugates─monoclonal antibodies conjugated to a cytotoxic moiety─are efficient therapeutics that can kill malignant cells overexpressing a target gene. Linking an antibody with radioisotopes (radioimmunoconjugates) enables powerful diagnostics and/or closely related therapeutic applications, depending on the isotope. To generate site-specific radioimmunoconjugates, we utilized genetic code expansion and subsequent conjugation by inverse electron-demand Diels-Alder cycloaddition reactions. We show that, using this approach, site-specific labeling of trastuzumab with either zirconium-89 (89Zr) for diagnostics or lutetium-177 (177Lu) for therapeutics yields efficient radioimmunoconjugates. Positron emission tomography imaging revealed a high accumulation of site-specifically 89Zr-labeled trastuzumab in tumors after 24 h and low accumulation in other organs. The corresponding 177Lu-trastuzumab radioimmunoconjugates were comparably distributed in vivo.

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Conflict of interest statement

The authors declare the following competing financial interest(s): Koehler, Sauter, Lemke and VERAXA Biotech GmbH holds patents regarding the site specific modification of antibodies.

Figures

Figure 1
Figure 1
General scheme of antibody labeling via SPIEDAC and chemical structures of compounds 1–3. (A) Principle of SPIEDAC of site-specifically introduced trans-cyclooctene amino acid in an antibody with tetrazine payload. (B) N6-({[(S,E)-Cyclooct-2-en-1-yl]oxy}carbonyl)-l-lysine (TCO*A, 1). (C) DFO-tetrazine (2). (D) DOTA-PEG9-tetrazine (3).
Figure 2
Figure 2
Pharmacokinetics and PET imaging of 89Zr-trastuzumabA121DFO-Tet. (A) PET/MR imaging at 24, 72, and 168 h after injection of 4 MBq 89Zr-trastuzumabA121DFO-Tet. Shown are transaxial, sagittal, and coronal fused images and the maximum intensity projection of the PET. (B) Ex vivo biodistribution at 168 h (n = 4). (C) Uptake values (%ID/cc) in tumor, blood, and liver after 24, 72, and 168 h.
Figure 3
Figure 3
Biodistribution of 177Lu-trastuzumabA121PEG9-DOTA. Ex vivo biodistribution at 48 h (n = 3); one control animal received 177Lu-DOTA-PEG9-tetrazine.
See this image and copyright information in PMC

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