Cherif Boutheina, Roget André, Villiers Christian L, Calemczuk Roberto, Leroy Vincent, Marche Patrice N, Livache Thierry, Villiers Marie-Bernadette
Laboratoire d'Immunochimie, Commissariat à L'Energie Atomique-Grenoble/Départment Réponse et Dynamique Cellulaire, Institut National de la Santé et de la Recherche Médicale U548, Université J. Fourier, Grenoble, France.
Clin Chem. 2006 Feb;52(2):255-62. doi: 10.1373/clinchem.2005.058727. Epub 2005 Dec 8.
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.
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.
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.
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.
