Ca. Wijayawardhana et al., Micro volume rotating disk electrode (RDE) amperometric detection for a bead-based immunoassay, ANALYT CHIM, 399(1-2), 1999, pp. 3-11
The application of the rotating disk electrode (RDE) directly on a micro dr
op of 50 mu l has been developed for electrochemical detection in a bead-ba
sed immunoassay for the model analyte mouse IgG. The enzyme label alkaline
phosphatase (ALP) was used for enzymatic turnover of the substrate 4-aminop
henyl phosphate (PAPP) to the electroactive product 4-aminophenol (PAP), Th
e determination of 4-aminophenol by the RDE has a linear range from 1.9 x 1
0(-8) to 1.9 x 10(-4) M. Direct analysis in a drop minimizes enzyme product
dilution, a key requirement in achieving very low limits of detection. Fur
thermore, the detection scheme allows enzyme turnover to be monitored conti
nuously, thus reducing errors associated with obtaining data at fixed times
as in flow injection analysis. An unoptimized calibration curve for mouse
IgG was obtained from 50 to 5000 ng/ml. (C) 1999 Elsevier Science B.V. All
rights reserved.