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Proceedings Paper

Sensitive detection of p53 antibodies in a homogeneous fluorescence assay format
Author(s): Hannes Neuweiler; Andreas Schulz; Juergen M. Wolfrum; Markus Sauer
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Paper Abstract

Circulating p53 autoantibodies are found to be a universal and highly specific tumor marker for malignant diseases. Hence, sereological screening for p53 autoantibodies at low concentration levels has become increasingly relevant for early-stage and follow-up of tumor diagnostics. We developed a new method for the highly sensitive detection of p53 antibodies in a homogeneous fluorescence assay format. Short, linear peptide derived form antibody recognition sequences so human p53 were labeled with an oxazine dye. Hydrophobic interactions constrain a conformation, where the dye interacts selectively with a tryptophan residue in the peptide sequence. Subsequently, the fluorescence of the dye is quenched efficiently due to electron transfer from the indole derivative to the dye in the excited state. Specific antibody recognition induces a conformational change in the peptide structure, repealing the dye-tryptophan interaction. Consequently, a fluorescence increase upon antibody binding signals the binding event. The long-wavelength absorption and emission characteristics of the probe and the use of a red pulsed diode laser as excitation source in a confocal fluorescence microscopic set-up allows ultra sensitive antibody detection at the single-molecule level. The effectiveness of the probes are highlighted by the detection of individual p53 autoantibodies directly in serum dilutions of cancer patients.

Paper Details

Date Published: 21 June 2002
PDF: 9 pages
Proc. SPIE 4626, Biomedical Nanotechnology Architectures and Applications, (21 June 2002); doi: 10.1117/12.472093
Show Author Affiliations
Hannes Neuweiler, Ruprecht-Karls-Univ. Heidelberg (Germany)
Andreas Schulz, Ruprecht-Karls-Univ. Heidelberg (Germany)
Juergen M. Wolfrum, Ruprecht-Karls-Univ. Heidelberg (Germany)
Markus Sauer, Ruprecht-Karls-Univ. Heidelberg (Germany)


Published in SPIE Proceedings Vol. 4626:
Biomedical Nanotechnology Architectures and Applications
Raymond P. Mariella; Catherine J. Murphy; Michelle Palmer; David A. Dunn; Darryl J. Bornhop; David A. Dunn; Raymond P. Mariella; Catherine J. Murphy; Dan V. Nicolau; Shuming Nie; Michelle Palmer; Ramesh Raghavachari; Darryl J. Bornhop; Ramesh Raghavachari; Shuming Nie; Ramesh Raghavachari, Editor(s)

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