Competitive immunoassay for vancomycin using capillary electrophoresis with laser-induced fluorescence detection.
A competitive immunoassay using capillary electrophoresis with laser-induced fluorescence was developed for vancomycin. Capillary electrophoresis using a Tris-glycine running buffer provided adequate separation of the antibody-bound from the unbound fluorescent probe (tracer) in less than 4 min. Laser-induced fluorescence polarization (LIFP) provided high sensitivity detection and simultaneous monitoring of fluorescence intensity and polarization. A fluorescence polarization value of 0.30 confirmed the formation of the antibody-tracer complex. Calibration curves showed a working linear range of 2-3 orders of magnitude with a minimum detectable concentration of 0.98 ng mL(-1) (or 1.1 fg vancomycin). Clinical samples obtained from patients undergoing vancomycin treatment were analyzed for vancomycin and the results correlated well with a standard immunoassay based on latex particle detection that was routinely used by a hospital laboratory. Only 1/10 of the reagents were needed as compared with the standard immunoassay.
Lam
Michael T
MT
Environmental Health Sciences Program, Department of Public Health Sciences, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, AB, Canada T6G 2G3.
Le Chris
X
X
D000900:Anti-Bacterial Agents / Q000097:blood*; D019075:Electrophoresis, Capillary / Q000379:methods; D005454:Fluorescence Polarization; D006801:Humans; D007118:Immunoassay / Q000379:methods; D014640:Vancomycin / Q000097:blood*
D000900:Anti-Bacterial Agents; D014640:Vancomycin