title: Antibody detection in human serum using a versatile protein chip platform constructed by applying nanoscale self-assembled architectures on gold.
abstract: We report a novel high-throughput (HTP) protein chip platform, constructed on gold using self-assembly techniques, for conducting high quality antigen-antibody interactions. Biotinylated monolayers were used to immobilize a streptavidin surface with high packing density. This biocompatible platform was then used for detection of serum IgM antibodies. Serum samples of patients suspected to suffer from Lyme borreliosis were used to validate the protein chip platform using biotinylated peptide AAOspC8 molecules as the test probes. Various experimental parameters such as the effect of concentration of probes, targets, temperature of incubation, and their effect on the resulting signal-to-noise ratio are described in detail. Highly specific protein interaction data with a high signal-to-noise ratio were obtained with serum sample solutions as low as 1 microL/spot (1/10 diluted).
authors.affiliation: ThermoHybaid, Sedanstrasse 18, 890 77 Ulm, Germany. pavlickova@thermohybaid.de
authors.forename: Petra
authors.initials: P
authors.lastname: Pavlickova
authors.forename: Nina Mejlhede
authors.initials: NM
authors.lastname: Jensen
authors.forename: Hubert
authors.initials: H
authors.lastname: Paul
authors.forename: Michael
authors.initials: M
authors.lastname: Schaeferling
authors.forename: Chiara
authors.initials: C
authors.lastname: Giammasi
authors.forename: Margit
authors.initials: M
authors.lastname: Kruschina
authors.forename: Wei-Dong
authors.initials: WD
authors.lastname: Du
authors.forename: Michael
authors.initials: M
authors.lastname: Theisen
authors.forename: Michael
authors.initials: M
authors.lastname: Ibba
authors.forename: Flavio
authors.initials: F
authors.lastname: Ortigao
authors.forename: Dev
authors.initials: D
authors.lastname: Kambhampati
mesh_terms: D000907:Antibodies, Bacterial / Q000097:blood*; D000941:Antigens / Q000378:metabolism; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins / Q000276:immunology; D001672:Biocompatible Materials; D025065:Borrelia burgdorferi / Q000276:immunology; D006046:Gold / Q000737:chemistry*; D006801:Humans; D007075:Immunoglobulin M / Q000097:blood*; D008193:Lyme Disease / Q000276:immunology; D036103:Nanotechnology; D040081:Protein Array Analysis / Q000379:methods*; D011485:Protein Binding; D011487:Protein Conformation; D013696:Temperature; D013997:Time Factors
chemical_list: D000907:Antibodies, Bacterial; D000941:Antigens; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D001672:Biocompatible Materials; D007075:Immunoglobulin M; C079616:OspC protein; D006046:Gold