Detection and Discrimination of PrPScby Multi-spectral Ultraviolet Fluorescence

Quantum dots and prion proteins

“…In order to avoid the use of antibodies as mentioned in previous paragraph, several spectroscopic methods, such as multispectral UV fluoroscopy, 54 fluorescence correlation spectroscopy, 55,56 magnetic resonance spectroscopy [57][58][59] or Fourier-transformed infrared spectroscopy (FTIR) 60,61 have been employed. The main disadvantages of these methods include requirements of expensive and sophisticated equipment as well as skilled operator.…”

Prion Protein and the Molecular Features of Transmissible Spongiform Encephalopathy Agents

“…However, it is important to point out that since disease-associated PrP is found in many different forms, a collection of conformational states and entities will fall under these terms. TSE-associated PrP molecules can vary in resistance to proteolysis (Bessen and Marsh 1994;Tagliavini et al 1994;Bessen et al 1995;Safar et al 1998;Caughey et al 1998a;Tzaban et al 2002;Horiuchi et al 2002), insolubility in detergents (Somerville et al 1989;Bessen and Marsh 1994;Muramoto et al 1996), secondary structure (Caughey et al 1998a), glycoform ratios (Kascsak et al 1986;Somerville and Ritchie 1990;Monari et al 1994;Collinge et al 1996;Somerville et al 1997;Rudd et al 1999;Hope et al 1999;Race et al 2002), exposure of epitopes and conformational stability in denaturants (Safar et al 1998;Peretz et al 2001a;Safar et al 2002), multi-spectral ultraviolet fluorescence Prion Protein and the Molecular Features of Transmissible Spongiform 13 (Rubenstein et al 1998), ultrastructure (McKinley et al 1991Giaccone et al 1992;Jeffrey et al 1994) and membrane topology (Hegde et al 1999). Complicating matters further is the fact noted above that Prp sc can be isolated as a complex of both partially PK-resistant and fully PK-sensitive molecules .…”

“…The existence of TSE strains independent of the host PrP genotype presents a major challenge to the protein-only hypothesis for the TSE agent. However, there is evidence to support the hypothesis that the mechanism for TSE strain propagation is determined by the self-propagation of Prp res molecules that differ in conformation, GdnHCI unfolding, glycoform ratios, polymeric states, and/or ligand associations (Bessen and Marsh 1994;Bessen et al 1995;Collinge et al 1996;Telling et al 1996;Somerville et al 1997;Safar et al 1998;Caughey et al 1998a;Rubenstein et al 1998;Peretz et al 2001aPeretz et al , 2002.…”

“…Wadsworth and colleagues have shown that metal ion binding can influence the site of PK cleavage in Prp res (Wadsworth et al 1999). Evidence for multiple strain differences in Prp sc conformation has also been obtained with multispectral ultraviolet fluorescence spectroscopy (Rubenstein et al 1998), denaturant-induced PK-susceptibility (Peretz et al 2001a), and by comparing antibody epitope exposure caused by denaturant treatments, a technique called conformation-dependent immunoassay (CD!) (Safar et al 1998(Safar et al , 2002.…”

Detection of prion protein using a capillary electrophoresis-based competitive immunoassay with laser-induced fluorescence detection and cyclodextrin-aided separation

“…Some of these tests have been commercialized and are used to test animals post mortem. Research approaches to develop preclinical assays include a variety of techniques including RNA aptamer technology [11], spectroscopy-based methods [12][13][14], protein-misfolding cyclic amplification [15], and capillary electrophoresis (CE) immunoassay [16]. An ideal diagnostic test should be (i) rapid and high-throughput, (ii) accurate, sensitive, and specific, (iii) robust, and (iv) using a sample from readily available tissues.…”