Characterization of 111In and 177Lu-labeled antibodies binding to CD44v6 using a novel automated radioimmunoassay.
Targeted cancer therapies rely on bifunctional molecules, typically a protein that specifically recognizes tumor cells and a toxic component which is linked to the protein. Therefore, development of such therapies includes detailed characterizations of protein-cell interactions in order to find a good targeting agent. Knowledge of factors such as antibody-antigen specificity, as well as cellular uptake, retention and affinity of the antibody are necessary in order to be successful. In this paper, we have used a novel instrument, LigandTracer Yellow, to characterize the interactions of (111)In and (177)Lu-labeled monoclonal antibodies (MAbs) with CD44v6. Uptake studies with varying specific radioactivity of the chimeric MAb U36 and with an irrelevant antibody for the CD44v6 receptor verified the reliability of the method, as well as the specificity of the antibody-receptor binding. Uptake, retention, and affinity were very similar for the (111)In and (177)Lu-labeled conjugate, and were in line with earlier studies using manual methods. The fact that no adverse effects from labeling were seen, together with the high retention, could make these conjugates promising candidates for imaging and therapy of certain cancer types in the future. The novel LigandTracer technology reduced the workload and reagent spending while providing data with superior time resolution. The obtained results were in agreement with previously reported findings. In addition the real-time detection and higher time resolution made more detailed studies of the interactions possible.
Nestor
Marika
M
Unit of Otolaryngology and Head & Neck Surgery, Department of Surgical Sciences, Uppsala University, Uppsala, Sweden. marika.nestor@bms.uu.se
Andersson
Karl
K
Lundqvist
Hans
H
D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism*; D001331:Automation; D045744:Cell Line, Tumor; D006801:Humans; D018960:Hyaluronan Receptors / Q000276:immunology*; D007205:Indium Radioisotopes; D007553:Isotope Labeling; D008187:Lutetium / Q000378:metabolism*; D011863:Radioimmunoassay / Q000379:methods*
D000911:Antibodies, Monoclonal; C112697:CD44v6 antigen; D018960:Hyaluronan Receptors; D007205:Indium Radioisotopes; D008187:Lutetium