Surround optical fiber immunoassay (SOFIA): an ultra-sensitive assay for prion protein detection.
We describe the development of a new technology (SOFIA) and demonstrate its utility by establishing a sensitive and specific assay for PrP(Sc). SOFIA is a surround optical fiber immunoassay which is comprised of a set of specific monoclonal antibodies and comprehensive capture of high energy fluorescence emission. In its current format, this system is capable of detecting less than 10 attogram (ag) of hamster, sheep and deer recombinant PrP. Approximately 10 ag of PrP(Sc) from 263 K-infected hamster brains can be detected with similar lower limits of PrP(Sc) detection from the brains of scrapie-infected sheep and deer infected with chronic wasting disease. These detection limits allow protease treated and untreated material to be diluted beyond the point where PrP(C), non-specific proteins or other extraneous material may interfere with PrP(Sc) signal detection and/or specificity. This not only eliminates the issue of specificity of PrP(Sc) detection but also increases sensitivity since the possibility of partial PrP(Sc) proteolysis is no longer a concern. SOFIA will likely lead to early antemortem detection of transmissible encephalopathies and is also amenable for use with additional target amplification protocols. SOFIA represents a sensitive means for detecting specific proteins involved in disease pathogenesis and/or diagnosis that extends beyond the scope of the transmissible spongiform encephalopathies.
Chang
Binggong
B
Department of Biochemistry, SUNY Downstate Medical Center, 450 Clarkson Avenue, Brooklyn, NY 11203, USA.
Gray
Perry
P
Piltch
Martin
M
Bulgin
Marie S
MS
Sorensen-Melson
Sharon
S
Miller
Michael W
MW
Davies
Paul
P
Brown
David R
DR
Coughlin
Daniel R
DR
Rubenstein
Richard
R
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015153:Blotting, Western; D001921:Brain / Q000276:immunology; D006224:Cricetinae; D003670:Deer; D005453:Fluorescence; D007118:Immunoassay / Q000379:methods*; D055100:Optical Fibers; D018620:PrPSc Proteins / Q000276:immunology / Q000302:isolation & purification*; D012608:Scrapie / Q000175:diagnosis*; D012680:Sensitivity and Specificity; D012756:Sheep; D034081:Wasting Disease, Chronic / Q000175:diagnosis*
D000911:Antibodies, Monoclonal; D018620:PrPSc Proteins