Journal: PLoS ONE
Article Title: An Integrated Peptide-Antigen Microarray on Plasmonic Gold Films for Sensitive Human Antibody Profiling
doi: 10.1371/journal.pone.0071043
Figure Lengend Snippet: A) Peptide-antigen microarray layout including modified and unmodified histone H2A, H2B, H3, H4 peptides as well as whole antigens. Ac: acetylated; aa: amino acid; Me1: methylated; Me2: dimethylated; Me3: trimethylated; Ph: phosphorylated; K: Lysine; S: Serine. Number indicates amino acid position from the N-terminus of its corresponding histone proteins. Peptide sequences are listed in . B) Microarray imaging results of a SLE patient serum probed on a plasmonic gold substrate (left) vs. on a commercial streptavidin-glass substrate (right) shown in the same IRDye800 fluorescence intensity scale. C) Dynamic range of fluorescence signal reading among 20 SLE patients measured on each peptide and antigen spots on avidin/gold slide (upper panel) and commercial streptavidin/glass slide (lower panel). Each dot represents a fluorescence intensity measured on a peptide or antigen spot shown along the x-axis in the serum of each of the 20 SLE patients. A dot is drawn only when the fluorescence intensity is above the background by 2 standard deviation of the background signal.
Article Snippet: The avidin coated gold slides (or commercial streptavidin coated glass slides) were loaded into a microarray printing robot (Bio-Rad) where 0.2 mg/ml biotin conjugated peptide in PBS supplemented with 4% glycerol was printed using solid pins (Arrayit) at 25°C and 60% humidity, resulting in microarray feature diameters of ∼200 μm.
Techniques: Microarray, Modification, Methylation, Imaging, Fluorescence, Avidin-Biotin Assay, Standard Deviation