Construction of Nucleic Acid antibodies to PrP(sc)

DESCRIPTION (Applicant's abstract): We propose to develop a method for generating amplifiable nucleic acid (NA) antibodies specific for the infectious particle responsible for prion disease, PrP(sc). NA antibodies have the ability to bind proteins with high affinity and specificity. Amplifiable NA antibodies will be generated by using a variation of the SELEX procedure that takes advantage of the activity of Q-beta replicase which replaces three steps in SELEX (reverse transcription, PCR amplification and RNA transcription). Moreover, the selected sequence that encodes the protein specific binding element is contained within an RNA vector derived from a Q-beta replicase template (MNV), providing the means for exponential amplification. This new method greatly simplifies and improves upon the SELEX procedure, enabling its application in the common molecular biology laboratory. Accumulation of PrP(sc) in the brain leads to untreatable spongiform encephalitis, typically acquired through consumption of contaminated beef. Since current tests for prions takes years, and since the number of cases in the United States is on the rise, there is an urgent need for an inexpensive, rapid diagnostic assay. NA antibodies specific for PrP(sc) that are amplifiable by Q-beta replicase are ideal for the development of a highly sensitive diagnostic assay using the Company's core, proprietary technology. PROPOSED COMMERCIAL APPLICATION: Our selection process may lead to a NA antibody production service, the development of kit for sale for research purposes or the creation of a market for the enzyme, Q-beta replicase. Amplifiable NA antibodies for PrP(sc) can be used as the raw material to develop an inexpensive diagnostic assay.