PHARMACOKINETICS BY FLOW CYTOMETRY: RECOMMENDATIONS FOR DEVELOPMENT AND VALIDATION OF FLOW CYTOMETRIC METHOD FOR PHARMACOKINETIC STUDIES - Flow Cytometry in Drug Discovery and Development - page 226

FIGURE 11.3 Drug X standard curve. A polyclonal antibody drug X dose-response is
shown. Standard reference material was spiked at difference concentrations into pooled normal
human serum. Standard curve is illustrated as concentration ( g/mL, x-axis) versus binding

signal (MFI median, y-axis). Data were collected on FACSCCalibur

m

.

11.2.7 Assay Development Study
11.2.7.1 General Considerations Most considerations relevant to general assay
development practices would apply to flow-based assays. During assay development,
the following variables are typically examined to define the optimal conditions: cell
number, drug concentration across the standard curve range, cell-drug incubation
time/temperature/wash, cell-bound drug secondary detection Ab incubation time and

FIGURE 11.4 Drug Y standard curve. A humanized IgG1 monoclonal antibody drug Y
dose-response is shown. Standard reference material was spiked at difference concentrations
into pooled normal human serum. Standard curve is illustrated as concentration ( m g/mL, x-axis)
versus binding signal (MFI median, y-axis). Data were collected on FACSCanto II.