Surround optical fiber immunoassay (SOFIA): An ultra-sensitive assay for prion protein detection

• Published in: Journal of virological methods 2009-07, Vol.159 (1), p.15-22
• Main Authors: Chang, Binggong, Gray, Perry, Piltch, Martin, Bulgin, Marie S., Sorensen-Melson, Sharon, Miller, Michael W., Davies, Paul, Brown, David R., Coughlin, Daniel R., Rubenstein, Richard
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Monoclonal - immunology; Biological and medical sciences; Blotting, Western; Brain - immunology; Cricetinae; Deer; Fiber optics; Fluorescence; Fundamental and applied biological sciences. Psychology; Immunoassay; Immunoassay - methods; Microbiology; Optical Fibers; PrP; PrPSc Proteins - immunology; PrPSc Proteins - isolation & purification; Scrapie - diagnosis; Sensitivity and Specificity; Sheep; SOFIA; Techniques used in virology; Virology; Wasting Disease, Chronic - diagnosis
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2009.02.019

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.