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.
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
Evgenia G
EG
Matveeva
Zygmunt
Z
Gryczynski
Joseph R
JR
Lakowicz
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