Radioimmunoconjugates for theragnostics of Prostate Stem Cell Antigen (PSCA)-expressing tumors

Aim: Advances in molecular engineering have led to the development of a multiplicity of antibody fragments with variations in molecular size. With respect to tumor targeting, the molecular size evidently determines the tumor uptake and pharmacokinetics. Consequently, they are proposed for different applications: small radiolabeled antibody fragments, such as single-chain variable fragments (scFv, 25-35 kDa) for tumor imaging and large full-size monoclonal antibodies (mAbs, 150 kDa) for radioimmunotherapy. Here, mAbs and thereof derived scFvs were produced that are directed against the prostate stem cell antigen (PSCA). Due to its overexpression on the surface of various tumor types, including prostate cancer and its metastases, it is proposed as a promising tumor target structure. Both antibody-based targeting molecules might provide a combinatory tool for theranostics of PSCA-positive prostate cancer.

Methods: In this study, two different anti-PSCA mAb clones, RD1 and RD2, as well as their respective anti-PSCA scFvs were produced and compared with regard to their binding properties towards PSCA, using flow cytometry analysis. The anti-PSCA mAbs were conjugated with the chelating agent p-SCN-CHX-A’’-DTPA, measured by MALDI-TOF, and subsequently radiolabeled with lutetium-177, whereas the scFvs were radiolabeled with technetium-99m on their histidine-tag. Thereafter, all radiolabeled conjugates were characterized by thin-layer chromatography, and regarding binding properties on PC3-PSCA cells in vitro.

Results: The non-radiolabeled anti-PSCA mAbs RD1 and RD2 showed a high affinity, with dissociation constants of 10 and 6 nM, respectively. The corresponding scFvs of RD1 and RD2 exhibit a lower affinity, with Kd-values of 170 and 98 nM. Both full mAbs were conjugated with about three CHX-A’’-DTPA. This conjugation had no influence on binding affinity towards the PSCA. Subsequent radiolabeling of the mAb-conjugates and scFvs could be performed with high radiochemical purity (> 95%) with preserving their binding properties to the PSCA.

Conclusion: Full-size mAbs and scFvs that target the tumor antigen PSCA were successfully produced and radiolabeled. The in vitro characterization showed promising results to proceed with studies on tumor mouse models.