NR AWHT
AU Kamali-M,M.; Schallmeiner,E.; Englund,H.; Ekholm Pettersson,F.; Söderberg,O.; Sehlin,D.; Lord,A.; Nilsson,L.; Lannfelt,L.; Landegren,U.
TI Sensitive proximity ligation-based detection of biomarkers of protein folding disorders
QU International Conference - Prion 2006: Strategies, advances and trends towards protection of society - 3.10.-6.10.2006, Torino, Italy, Lingotto Conference Centre - Poster sessions DIA-29
PT Konferenz-Poster
AB The development of sensitive and specific techniques for protein detection could provide opportunities for early detection and therapeutic intervention in protein folding disorders, for instance prion and Alzheimer's diseases. The proximity ligation assay (PLA) is a recently developed method in which specific proteins are analyzed by translating detection signals to DNA sequences. In this method target molecules are recognized by pairs of affinity probes - e.g. antibodies. Proximity probe pairs, used for detection are prepared by attaching DNA strands to the affinity probes. When a pair of such probes bind to a common target molecule, the free ends of the attached oligonucleotides are brought in proximity and can be hybridized to a connector oligonucleotide, allowing the ends to be joined by enzymatic DNA ligation and detected by PCR. We have applied PLA for sensitive identification of peptide oligomers, while excluding detection of protein monomers. Soluble A oligomers, which have been shown to mediate neurotoxicity, may be used as a candidate biomarker for diagnosis of Alzheimer's disease at early stages. The combination of efficient PCR amplification and the use of two or more binding reagents provide very high sensitivity and specificity of detection, surpassing conventional protein detection methods. Furthermore, the proximity ligation technique can be carried out as an in situ assay, as in the homogenous assay - requiring very small amount of materials to be tested -, or in a solid-phase format in which the target molecules to be detected are immobilized on a surface using affinity probes, while other materials are washed away. Proximity ligation can, therefore, provide a powerful molecular tool for studying the biology of protein folding disorders and for early diagnosis.
AD M. Kamali-M., E. Schallmeiner, O. Söderberg, U. Landegren: Department of Genetics and Pathology / Molecular Medicine, Uppsala University, Sweden; H. Englund, F. Ekholm Pettersson, D. Sehlin, A. Lord, L. Nilsson, L. Lannfelt: Department of Public Health and Caring Sciences / Section of Molecular Geriatrics; Dag Hammarskjölds väg 20, SE-75185 Uppsala, Sweden. E-mail: masood.kamali@genpat.uu.se
SP englisch
PO Italien