Fluorescent Immunoassay Development for PrP(Sc) Detection and Antemortem Diagnosis of TSEs
Abstract
The overall goal of our study is to develop methods of high-sensitivity and high-specificity for the antemortem diagnosis of prion diseases by detecting PrPsc in biological fluids using fluorescent immunoassay. During the first year of this contract, we have focused on improving the assay sensitivity by means of new biological protocols and hardware improvement. Concentration of PrP prior to the assay using sodium phosphotungstate treatment was shown to be compatible with the LIFIA and effective in improving the sensitivity. Use of PrPsc-specific reagents such as antibodies against advanced glycosylation end products (ACE) and DNA provided promising additional approaches for the development of high sensitivity immunoassay. Direct detection of blood PrPsc by Western blot following concentration of PrPc by anti-PrP antibody-coated beads improved the potential for detection of PrPsc in blood. These studies have provided important advancement in our capacity to effectively pursue a specific, sensitive PrPsc assay in easily obtainable biological fluids and will further our efforts toward the goal of a fluorescent immunoassay system using a laser-induced fluorescence spectrofluorometer.
Document Details
- Document Type
- Technical Report
- Publication Date
- Oct 01, 2004
- Accession Number
- ADA430442
Entities
People
- Richard I. Carp
Organizations
- Research Foundation For Mental Hygiene