Sensitive detection of p53 antibodies in a homogeneous fluorescence assay format

Hannes Neuweiler; Andreas Schulz; Juergen M. Wolfrum; Markus Sauer

doi:10.1117/12.472093

21 June 2002 Sensitive detection of p53 antibodies in a homogeneous fluorescence assay format

Hannes Neuweiler, Andreas Schulz, Juergen M. Wolfrum, Markus Sauer

Proceedings Volume 4626, Biomedical Nanotechnology Architectures and Applications; (2002) https://doi.org/10.1117/12.472093
Event: International Symposium on Biomedical Optics, 2002, San Jose, CA, United States

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.

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Hannes Neuweiler, Andreas Schulz, Juergen M. Wolfrum, and Markus Sauer "Sensitive detection of p53 antibodies in a homogeneous fluorescence assay format", Proc. SPIE 4626, Biomedical Nanotechnology Architectures and Applications, (21 June 2002); https://doi.org/10.1117/12.472093

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KEYWORDS

Luminescence

Tumors

Absorption

Biomedical optics

Cancer

Confocal microscopy

Diagnostics

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