NR AXWR
AU Snejdarkova,M.; Svobodova,L.; Polohova,V.; Hianik,T.
TI Detection of Prion Proteins in Blood Plasma Using Thickness Shear Mode Method and Amplification by Gold Nanoparticles
QU International Conference - Prion 2007 (26.-28.9.2007) Edinburgh International Conference Centre, Edinburgh, Scotland, UK - Book of Abstracts: Epidemiology, Risk Assessment and Transmission P04.34
IA http://www.prion2007.com/pdf/Prion Book of Abstracts.pdf
PT Konferenz-Poster
AB
Ante mortem detection of prion proteins is one of the most important problems in current diagnostics of prion diseases. Considering rather small concentration of cellular prions (PrPc) as well as pathogenic PrPsc the detection limit down to 1 pM should be achieved. The biosensors based on antibodies may be one of the alternative to current time consuming and expensive conventional tests based on Western Blotting. Here we present relative simple method of detection prion proteins in a blood plasma based on thickness shear mode method and amplification of the detection using gold nanoparticles. The antibodies against PrPc were immobilized on a surface of AT-cut quartz crystal by means of poly(amidoamine) (PAMAM) dendrimers of fourth generation (G4). The blood plasma of health individuals was allowing to flow through the microcell of 100 ml volume resulted decrease of series resonant frequency, fs, and increase of motional resistance, Rm. This evidence on binding the prions to the sensor surface and in increase of the mass of the biolayers. The increase of motional resistance suggests an increase of molecular slip between the biolayers and surrounding liquid. As soon as the measured parameters were stabilized the antibodies conjugated with gold nanoparticles were added, which resulted additional, more substantial decrease of series resonant frequency. This amplification effect allowed us to determine the PrPc with accuracy comparable to Western Blotting method.
Acknowledgements This work was financially supported by European Commission under 6 FP program NoE NeuroPrion (Contract No. FOOD-CT-2004-506579), by Agency for Promotion Research and Development under the contract No. APVV-20-P01705 and by Slovak Grant Agency (Projects No. 2/7134/7 to M.S. and 1/4016/07 to T.H)
AD M. Snejdarkova, L. Svobodova, V. Polohova, Institute of Animal Biochemistry and Genetics, Slovak Republic; T. Hianik, Comenius University, Slovak Republic
SP englisch
PO Schottland