title: Clinically related protein-peptide interactions monitored in real time on novel peptide chips by surface plasmon resonance imaging.
abstract: 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.
authors.affiliation: 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.
authors.forename: Boutheina
authors.initials: B
authors.lastname: Cherif
authors.forename: André
authors.initials: A
authors.lastname: Roget
authors.forename: Christian L
authors.initials: CL
authors.lastname: Villiers
authors.forename: Roberto
authors.initials: R
authors.lastname: Calemczuk
authors.forename: Vincent
authors.initials: V
authors.lastname: Leroy
authors.forename: Patrice N
authors.initials: PN
authors.lastname: Marche
authors.forename: Thierry
authors.initials: T
authors.lastname: Livache
authors.forename: Marie-Bernadette
authors.initials: MB
authors.lastname: Villiers
mesh_terms: D001666:Binding Sites, Antibody; D002624:Chemistry, Clinical; D006046:Gold; D016174:Hepacivirus / Q000276:immunology; D018937:Hepatitis C Antibodies / Q000737:chemistry*; D006801:Humans; D010455:Peptides / Q000737:chemistry*; D011108:Polymers / Q000737:chemistry*; D040081:Protein Array Analysis / Q000295:instrumentation / Q000379:methods*; D025941:Protein Interaction Mapping; D011758:Pyrroles / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000737:chemistry; D020349:Surface Plasmon Resonance
chemical_list: D018937:Hepatitis C Antibodies; D010455:Peptides; D011108:Polymers; D011758:Pyrroles; D011994:Recombinant Proteins; D012710:Serum Albumin, Bovine; C067635:polypyrrole; D006046:Gold