Antibody detection in human serum using a versatile protein chip platform constructed by applying nanoscale self-assembled architectures on gold.
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).
Pavlickova
Petra
P
ThermoHybaid, Sedanstrasse 18, 890 77 Ulm, Germany. pavlickova@thermohybaid.de
Jensen
Nina Mejlhede
NM
Paul
Hubert
H
Schaeferling
Michael
M
Giammasi
Chiara
C
Kruschina
Margit
M
Du
Wei-Dong
WD
Theisen
Michael
M
Ibba
Michael
M
Ortigao
Flavio
F
Kambhampati
Dev
D
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
D000907:Antibodies, Bacterial; D000941:Antigens; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D001672:Biocompatible Materials; D007075:Immunoglobulin M; C079616:OspC protein; D006046:Gold