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. CONCLUSION: 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.
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. CONCLUSION: 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.
Authors: Claude Nogues; Hervé Leh; Christopher G Langendorf; Ruby H P Law; Ashley M Buckle; Malcolm Buckle Journal: PLoS One Date: 2010-08-13 Impact factor: 3.240
Authors: Feliciana Real-Fernández; Irene Passalacqua; Elisa Peroni; Mario Chelli; Francesco Lolli; Anna Maria Papini; Paolo Rovero Journal: Sensors (Basel) Date: 2012-05-02 Impact factor: 3.576
Authors: Hong T T Huynh; Guillaume Gotthard; Jérome Terras; Gérard Aboudharam; Michel Drancourt; Eric Chabrière Journal: BMC Res Notes Date: 2015-06-24