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

TY - JOUR

T1 - Surround optical fiber immunoassay (SOFIA):

T2 - an ultra-sensitive assay for prion protein detection

AU - Chang, B

AU - Gray, Perry

AU - Piltch, M

AU - MS, Bulgin

AU - Sorensen-Melson, S

AU - MW, Miller

AU - Davies, P

AU - DR, Brown

AU - DR, Coughlin

AU - Rubenstein, R

PY - 2009/2/27

Y1 - 2009/2/27

N2 - We describe the development of a new technology (SOFIA) and demonstrate its utility by establishing a sensitive and specific assay for PrPSc. 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 PrPSc from 263 K-infected hamster brains can be detected with similar lower limits of PrPSc 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 PrPC, non-specific proteins or other extraneous material may interfere with PrPSc signal detection and/or specificity. This not only eliminates the issue of specificity of PrPSc detection but also increases sensitivity since the possibility of partial PrPSc 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.

AB - We describe the development of a new technology (SOFIA) and demonstrate its utility by establishing a sensitive and specific assay for PrPSc. 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 PrPSc from 263 K-infected hamster brains can be detected with similar lower limits of PrPSc 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 PrPC, non-specific proteins or other extraneous material may interfere with PrPSc signal detection and/or specificity. This not only eliminates the issue of specificity of PrPSc detection but also increases sensitivity since the possibility of partial PrPSc 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.

UR - http://europepmc.org/abstract/med/19442839

U2 - 10.1016/j.jviromet.2009.02.019

DO - 10.1016/j.jviromet.2009.02.019

M3 - Article

C2 - 19442839

SN - 0166-0934

VL - 159

SP - 15

EP - 22

JO - Journal of Virological Methods

JF - Journal of Virological Methods

IS - 1

ER -