Myoglobin immunoassay based on metal particle-enhanced fluorescence.
Enhanced fluorescence on silver island films (SIFs) is utilized to develop a sandwich-format immunoassay for the cardiac marker myoglobin (Myo). Myoglobin was first captured on surfaces coated with anti-Myo antibodies; the surface was then incubated with fluorescently labeled anti-Myo antibodies. The system was examined on glass surfaces and on SIFs. We observed the enhancement of the signal from SIFs in the range of 10-15-fold if compared to the signal from the glass substrate not modified with a SIF. A kinetic immunoassay for Myo on SIF-modified surface results in a decreased background signal. The initial results show that it is possible to detect Myoglobin concentrations below 50 ng/mL, which is lower than clinical cut-off for Myoglobin in healthy patients. We suggest the use of SIF-modified substrates for increasing the sensitivity of surface assays with fluorescence detection.
Matveeva
Evgenia G
EG
Center for Fluorescence Spectroscopy, University of Maryland at Baltimore Medical School, Department of Biochemistry and Molecular Biology, 725 West Lombard Street, Baltimore, MD 21201, USA. eva@cfs.umbi.umd.edu
Gryczynski
Zygmunt
Z
Lakowicz
Joseph R
JR
D000911:Antibodies, Monoclonal; D015415:Biomarkers; D005456:Fluorescent Dyes; D015200:Fluoroimmunoassay / Q000379:methods; D007700:Kinetics; D009211:Myoglobin / Q000032:analysis* / Q000276:immunology; D011485:Protein Binding; D012235:Rhodamines; D013194:Staining and Labeling / Q000379:methods
D000911:Antibodies, Monoclonal; D015415:Biomarkers; D005456:Fluorescent Dyes; D009211:Myoglobin; D012235:Rhodamines