Clin Nucl Med. 2024 Apr 5. doi: 10.1097/RLU.0000000000005208. Online ahead of print.

ABSTRACT

PURPOSE: A monoclonal antibody, trastuzumab, is used for immunotherapy for HER2-expressing breast cancers. Large-sized antibodies demonstrate hepatobiliary clearance and slower pharmacokinetics. A trastuzumab fragment (Fab; 45 kDa) has been generated for theranostic use.

PATIENTS AND METHODS: Fab was generated by papain digestion. Trastuzumab and Fab have been radiolabelled with 177Lu after being conjugated with a bifunctional chelating. The affinity and target specificity were studied in vitro. The first-in-human study was performed.

RESULTS: The bifunctional chelating agent conjugation of 1-2 molecules with trastuzumab and Fab was detected at the molar ratio 1:10 in bicarbonate buffer (0.5 M, pH 8) at 37°-40°C. However, 2-3 molecules of bifunctional chelating agent were conjugated when DMSO in PBS (0.1 M, pH 7) was used as a conjugation buffer at a molar ratio of 1:10. The radiolabelling yield of DOTA-conjugated Fab and trastuzumab at pH 5, 45°C to 50°C, with incubation time 2.5-3 hours was 80% and 41.67%, respectively. However, with DOTAGA-conjugated trastuzumab and Fab, the maximum radiolabelling yield at pH 5.5, 37°C, and at 2.5-3 hours was 80.83% and 83%, respectively. The calculated Kd of DOTAGA Fab and trastuzumab with HER2-positive SKBR3 cells was 6.85 ± 0.24 × 10-8 M and 1.71 ± 0.10 × 10-8 M, respectively. DOTAGA-Fab and trastuzumab showed better radiolabelling yield at mild reaction conditions.177Lu-DOTAGA-Fab demonstrated higher lesion uptake and lower liver retention as compared with 177Lu-DOTAGA-trastuzumab. However, 177Lu-DOTAGA-Fab as compared with 177Lu-DOTAGA-trastuzumab showed a relatively early washout (5 days) from the lesion.

CONCLUSIONS: 177Lu-DOTAGA-Fab and trastuzumab are suitable for targeting the HER2 receptors.

PMID:38579266 | DOI:10.1097/RLU.0000000000005208