The binding affinity of [125I]I-Ec1-LoPE to SKOV3 and OVCAR3 cells was measured using LigandTracer Grey Instrument (Ridgeview Instruments, Vänge, Sweden) as described previously [51 (link)].
Briefly, one day before the experiment, cells were seeded in one sector of a Petri dish. The experiment was performed at room temperature. Two separate measurements were performed for each cell line. The kinetics of binding of [125I]I-Ec1-LoPE to the cells was measured continuously after adding the labelled compound to obtain concentrations of 11 and 33 nM. Thereafter, the cell culture medium was replaced by a medium not containing [125I]I-Ec1-LoPE and dissociation rate was measured. The equilibrium dissociation constants (KD) were calculated using the TraceDrawer Software (Ridgeview Instruments, Vänge, Sweden). Based on association and dissociation rate data, the equilibrium dissociation constants (KD) were calculated using the TraceDrawer Software (Ridgeview Instruments, Vänge, Sweden). Interaction heterogeneity was estimated using interaction map analysis (Ridgeview Diagnostics, Uppsala, Sweden).
Briefly, one day before the experiment, cells were seeded in one sector of a Petri dish. The experiment was performed at room temperature. Two separate measurements were performed for each cell line. The kinetics of binding of [125I]I-Ec1-LoPE to the cells was measured continuously after adding the labelled compound to obtain concentrations of 11 and 33 nM. Thereafter, the cell culture medium was replaced by a medium not containing [125I]I-Ec1-LoPE and dissociation rate was measured. The equilibrium dissociation constants (KD) were calculated using the TraceDrawer Software (Ridgeview Instruments, Vänge, Sweden). Based on association and dissociation rate data, the equilibrium dissociation constants (KD) were calculated using the TraceDrawer Software (Ridgeview Instruments, Vänge, Sweden). Interaction heterogeneity was estimated using interaction map analysis (Ridgeview Diagnostics, Uppsala, Sweden).
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