Preparation of ²¹²Pb-labeled monoclonal antibody using a novel ²²⁴Ra-based generator solution

Alpha-emitting radionuclides have gained considerable attention as payloads for cancer targeting molecules due to their high cytotoxicity. One attractive radionuclide for this purpose is ²¹²Pb, which by itself is a β-emitter, but acts as an in vivo generator for its short-lived α-emitting daughters. The standard method of preparing ²¹²Pb-labeled antibodies requires handling and evaporation of strong acids containing high radioactivity levels by the end user. An operationally easier and more rapid process could be useful since the 10.6h half-life of ²¹²Pb puts time constraints on the preparation protocol. In this study, an in situ procedure for antibody labeling with ²¹²Pb, using a solution of the generator nuclide ²²⁴Ra, is proposed as an alternative protocol for preparing ²¹²Pb-radioimmunoconjugates.

Radium-224, the generator radionuclide of ²¹²Pb, was extracted from its parent nuclide, ²²⁸Th. Lead-212-labeling of the TCMC-chelator conjugated monoclonal antibody trastuzumab was carried out in a solution containing ²²⁴Ra in equilibrium with progeny. Subsequently, the efficiency of separating the ²¹²Pb-radioimmunoconjugate from ²²⁴Ra and other unconjugated daughter nuclides in the solution using either centrifugal separation or a PD-10 desalting size exclusion column was evaluated and compared.

Radiolabeling with ²¹²Pb in ²²⁴Ra-solutions was more than 90% efficient after only 30min reaction time at TCMC-trastuzumab concentrations from 0.15mg/mL and higher. Separation of ²¹²Pb-labeled trastuzumab from ²²⁴Ra using a PD-10 column was clearly superior to centrifugal separation. This method allowed recovery of approximately 75% of the ²¹²Pb-antibody-conjugate in the eluate, and the remaining amount of ²²⁴Ra was only 0.9±0.8% (n=7).

The current work demonstrates a novel method of producing ²¹²Pb-based radioimmunoconjugates from a ²²⁴Ra-solution, which may be simpler and less time-consuming for the end user compared with the method established for use in clinical trials of ²¹²Pb-TCMC-trastuzumab.