Biomedical Engineering Reference
In-Depth Information
11
PHARMACOKINETICS BY FLOW
CYTOMETRY: RECOMMENDATIONS
FOR DEVELOPMENT AND
VALIDATION OF FLOW CYTOMETRIC
METHOD FOR PHARMACOKINETIC
STUDIES
Y UANXIN X U AND S USAN M. R ICHARDS
11.1
INTRODUCTION
Flow cytometric methods can be used for quantitative analysis of analytes in
biological matrices. This chapter will discuss development and validation of flow
cytometric methods to quantify protein drugs for pharmacokinetic (PK) analysis for
use in preclinical or clinical studies.
The assay principle is based on binding of a drug to target molecules expressed on
the cell surface. Depending on the drug characteristics, this binding can be antibody
(Ab)-antigen (Ag) or receptor-ligand mediated. Detection of the bound drug at
different levels is usually achieved by a secondary antidrug antibody conjugated to
florescent dye (FITC, PE, etc.). A dose-response curve (or standard curve), drug
concentration versus signal, can be generated and drug concentration in unknown
samples will then be quantified by interpolating the signal from the standard curve.
Signal is typically MFI (median fluorescence intensity) or MESF (molecules of
equivalent soluble fluorochrome) converted fromMFI using calibration beads such as
Quantum
beads with QuickCal (Bang's Laboratories, Inc., Fishers, IN, USA).
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