In vitro evaluation of the monoclonal antibody Cu-64-IgG M75 against human carbonic anhydrase IX and its in vivo imaging
Abstrakt
Specific oncology diagnostics requires new types of the selective radiophannaceuticals, particularly those suitable for the molecular PET imaging. The aim of this work is to present a new, specific PET-immunodiagnostic radiopharmaceutical based on the monoclonal antibody IgG M75 targeting human carbonic anhydrase IX labelled with Cu-64 (T-1/2 = 12.70 h) and its in vitro and in vivo evaluation.
The antibody IgG M75 was conjugated with a non-commercial copper-specific chelator "phosphinate" and then labelled with the positron emitter 64Cu. Stability of the labelled conjugated was tested in human serum.
The immunoreactivity of the labelled conjugate was evaluated in vitro on a suitable cell cultures of the colorectal carcinoma (HT-29) and its imaging properties were estimated in vivo on a mouse model with inoculated colorectal carcinoma HT-29 imaged on a mu PET/CT. The tested radioimmunoconjugate was obtained in a specific activity of 0.25-0.5 MBq/mu g.
In vitro uptake experiments revealed specific binding to the HT-29 cells (45 +/- 2.8% of the total added activity) and the measured K-D value was found to be 9.2 nM. Imaging clearly demonstrated significant uptake of the labelled monoclonal antibody in the tumour at 18 h post administration.
The radioimmunoconjugate Cu-64-PS-IgG M75 seems to be a suitable candidate for PET diagnostics of hypoxic tumours expressing human carbonic anhydrase IX.