On-chip micro-flow polystyrene bead-based immunoassay for quantitative detection of tacrolimus (FK506).
Tacrolimus (FK506) is a widely used immunosuppressant for preventing allograft rejection and the treatment of atopic dermatitis. FK506 necessitates therapeutic drug monitoring because of inter- and intrapatient variability and the lack of correlation between the administered dose and the blood concentration. Previous immunoassay-based methods required a relatively long assay time and troublesome liquid-handling procedures. In the present study, we aimed to establish a rapid monitoring method for FK506 determination by using a poly(dimethylsiloxane) (PDMS)-based microfluidic device. Polystyrene beads were coated with mouse anti-FK506 antibody and placed in the flow channel. As a competitive assay, sample solution was allowed to react in the flow channel. After the addition of the fluorogenic substrate, the fluorescent signal was observed under a microscope. As a result, the developed assay allowed a short detection time of approximately 15 min per each sample and a high sensitivity even by using only a single bead. The feasibility of performing a competitive assay using a PDMS-based antibody chip gives promising results over the existing immunoassay-based methods.
Department of Chemical Materials Science, School of Materials Science, Japan Advanced Institute of Science and Technology, 1-1 Asahidai, Tatsunokuchi, Nomigun, Ishikawa 923-1292, Japan. murakami@mpcrf.or.jp
Yoshihiro
Y
Murakami
Tatsuro
T
Endo
Shouhei
S
Yamamura
Naoki
N
Nagatani
Yuzuru
Y
Takamura
Eiichi
E
Tamiya
D000818:Animals; D004129:Dimethylpolysiloxanes; D016903:Drug Monitoring / Q000379:methods*; D005456:Fluorescent Dyes; D007118:Immunoassay / Q000295:instrumentation* / Q000379:methods / Q000592:standards; D008252:Macaca fascicularis; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D008856:Microscopy, Fluorescence; D008863:Microspheres; D011137:Polystyrenes; D012828:Silicones; D016559:Tacrolimus / Q000097:blood*
D004129:Dimethylpolysiloxanes; D005456:Fluorescent Dyes; D011137:Polystyrenes; D012828:Silicones; C013830:baysilon; D016559:Tacrolimus