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Clinical Chemistry 0: clinchem.2005.058727v1, 2005; 10.1373/clinchem.2005.058727
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Received on August 4, 2005
Accepted on November 7, 2005

Automation and Analytical Techniques

Clinically Related Protein-Peptide Interactions Monitored in Real Time on Novel Peptide Chips by Surface Plasmon Resonance Imaging

Boutheina Cherif 1, André Roget 2, Christian L. Villiers 1, Roberto Calemczuk 2, Vincent Leroy 3, Patrice N. Marche 1, Thierry Livache 2, Marie-Bernadette Villiers 3*

1 Laboratoire d'Immunochimie, CEA-Grenoble/DRDC, INSERM U548, Université J. Fourier, Grenoble, France
2 CREAB (CEA, CNRS, UJF), CEA-Grenoble/DRFMC, UMR 5819, Grenoble, France
3 Laboratoire d'Immunochimie, CEA-Grenoble/DRDC, INSERM U548, Université J. Fourier, Grenoble, France, and Département d'Hépato-Gastroentérologie, CHU de Grenoble, Grenoble, France

* To whom correspondence should be addressed. E-mail: immuno{at}dsvgre.cea.fr.

Background: Developing rapid, high-throughput assays for detecting and characterizing protein-protein interactions is a great challenge in the postgenomic era. We have developed a new method that allows parallel analysis of multiple analytes in biological fluids and is suitable for biological and medical studies.

Methods: This technology for studying peptide-antibody interactions is based on polypyrrole-peptide chips and surface plasmon resonance imaging (SPRi). We generated a chip bearing a large panel of peptide probes by successive electro-directed copolymerizations of pyrrole-peptide conjugates on a gold surface.

Results: We provide evidence that (a) the signal produced by antibody binding is highly specific; (b) the detected signal specifically reflects the antibody concentration of the tested solution in a dose-dependent manner; (c) this technique is appropriate for analyzing complex media such as undiluted sera, a novelty with respect to previous techniques; and (d) correlation between classic ELISA results and the SPRi signal is good (P = 0.008). We also validated this system in a medical model by detecting anti-hepatitis C antibodies in patient-derived sera.

Conclusions: Because of its characteristics (easy preparation of the peptide chip; high-throughput, label-free, real-time detection; high specificity; and low background), this technology is suitable for screening biological samples and for large-scale studies.


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[Abstract] [Full Text] [PDF]


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