@article{AndresenGrotzingerZarseetal.2006, author = {Andresen, Heiko and Grotzinger, Carsten and Zarse, Kim and Kreuzer, Oliver Johannes and Ehrentreich-F{\"o}rster, Eva and Bier, Frank Fabian}, title = {Functional peptide microarrays for specific and sensitive antibody diagnostics}, issn = {1615-9853}, doi = {10.1002/pmic.200500343}, year = {2006}, abstract = {Peptide microarrays displaying biologically active small synthetic peptides in a high-density format provide an attractive technology to probe complex samples for the presence and/or function of protein analytes. We present a new approach for manufacturing functional peptide microarrays for molecular immune diagnostics. Our method relies on the efficiency of site-specific solution-phase coupling of biotinylated synthetic peptides to NeutrAvidin (NA) and localized microdispensing of peptide-NA-complexes onto activated glass surfaces. Antibodies are captured in a sandwich manner between surface immobilized peptide probes and fluorescence-labeled secondary antibodies. Our work includes a total of 54 peptides derived from immunodominant linear epitopes of the T7 phage capsid protein, Herpes simplex virus glycoprotein D, c-myc protein, and three domains of the Human coronavirus polymerase polyprotein and their cognate mAbs. By using spacer molecules of different type and length for NA-mediated peptide presentation, we show that the incorporation of a minimum spacer length is imperative for antibody binding, whereas the peptide immobilization direction has only secondary importance for antibody affinity and binding. We further demonstrate that the peptide array is capable of detecting low-picomolar concentrations of mAbs in buffered solutions and diluted human serum with high specificity}, language = {en} } @article{AndresenGroetzingerZarseetal.2006, author = {Andresen, Heiko and Gr{\"o}tzinger, Carsten and Zarse, Kim and Birringer, Marc and Hessenius, Carsten and Kreuzer, Oliver Johannes and Ehrentreich-F{\"o}rster, Eva and Bier, Frank Fabian}, title = {Peptide microarrays with site-specifically immobilized synthetic peptides for antibody diagnostics}, issn = {0925-4005}, doi = {10.1016/j.snb.2005.07.033}, year = {2006}, abstract = {Peptide microarrays bear the potential to discover molecular recognition events on protein level, particularly in the field of molecular immunology, in a manner and with an efficiency comparable to the performance of DNA microarrays. We developed a novel peptide microarray platform for the detection of antibodies in liquid samples. The system comprises site-specific solution phase coupling of biotinylated peptides to NeutrAvidin, localized microdispensing of peptide-NeutrAvidin conjugates onto activated glass slides and a fluorescence immuno sandwich assay format for antibody capture and detection. Our work includes synthetic peptides deduced from amino acid sequences of immunodominant linear epitopes, such as the T7 phage capsid protein, Herpes simplex virus glycoprotein D, c-myc protein and three domains of the Human coronavirus 229E polymerase polyprotein. We demonstrate that our method produces peptide arrays with excellent spot morphology which are capable of specific and sensitive detection of monoclonal antibodies from fluid samples.}, language = {en} }