NR ATEW
AU Yang,W.C.; Schmerr,M.J.; Jackman,R.; Bodemer,W.; Yeung,E.S.
TI Capillary electrophoresis-based noncompetitive immunoassay for the prion protein using fluorescein-labeled protein a as a fluorescent probe
QU Analytical Chemistry 2005 Jul 15; 77(14): 4489-94
PT journal article
AB A novel CE-based noncompetitive immunoassay for prion protein (PrP) was established. Fluorescein isothiocyanate (FITC)-labeled protein A (FITC-PrA) was used as a fluorescent probe to tag monoclonal antibody through noncovalent binding of FITC-PrA to the Fc region of the antibody. The FITC-PrA-Ab was incubated with the analyte, prion protein, under optimized condition, forming the immunocomplex FITC-PrA-Ab-PrP. The complex was separated and analyzed by capillary zone electrophoresis. The addition of carboxymethyl-beta-cyclodextrin in the running buffer as dynamical coating reagent improved the reproducibility and the resolution. The complex was isolated in less than 1 min with theoretical plates of 3.8 x 10(4). Relative standard deviations of peak height and migration time for the complex were 3.46 and 1.48%, respectively. A linear relationship was established for the bovine recombinant prion protein (rPrP) concentration in the range from 0.2 to 2.0 mug/mL and the peak height. The correlation factor was r2 = 0.9969. The estimated detection limit for rPrP was approximately 6 ng/mL, which is 3 times the signal-to-noise ratio. The method was successfully applied for testing blood samples from scrapie-infected sheep.
IN Die Autoren berichten über die Entwicklung eines nichtkompetitiven Immunoassays auf der Basis der Kapillarelektrophorese und wollen damit eine Empfindlichkeit von 6 ng/ml erreichen.
AD Department of Chemistry, Iowa State University, Ames, Iowa 50011, Ames Laboratory USDOE, Iowa State University, Ames, Iowa 50011, USA
SP englisch
PO USA