Journal: bioRxiv

Article Title: Comprehensive profiling of antibodies against multiple infectious diseases in serum and the airway mucosa using synthetic peptide-based linear epitope microarrays

doi: 10.1101/462689

Figure Lengend Snippet: (A) An example slide in which immunoglobulins in sera from different children were assayed. Each microarray slide comprised 24 mini-arrays (shown using the white boxes). Two example mini-arrays are enlarged: in the first mini-array a sample obtained from a one-day old neonate was analysed while in the second, serum from a two-month old infant was analysed. On each mini-array, peptide epitopes were printed and serum/mucosal samples incubated. Antibodies in patient samples that bound to peptide epitopes were identified using a cocktail of two detection antibodies: anti human IgG conjugated to Alex Fluor (AF) 647 (red fluorescent signal) and anti-human IgA conjugated to AF-555(green fluorescence). Spots with a yellow hue indicate antigens which were bound by both IgA and IgG in patient samples. Each peptide epitope was printed in duplicate: the locations of a selection of duplicate antigen pairs are shown. (B) The distribution of fluorescence signals from control spots. Each mini-array contained a set of positive control spots (comprising of an anti-human IgG that bound to all IgG immunoglobulins in the patient sample irrespective of antigenic specificity) and negative control spots (microarray printing buffer). Almost all negative control spots did not yield a fluorescent signal - i.e. median fluorescent intensity (MFI)=0. The positive control spots on the other hand yielded much higher MFIs, with most spots resulting in MFIs >10,000.

Article Snippet: Peptides were printed onto the epoxy slides using a non-contact inkjet microarray printer (Arrayjet, Scotland).

Techniques: Microarray, Incubation, Fluorescence, Selection, Positive Control, Negative Control

Journal: bioRxiv

Article Title: Comprehensive profiling of antibodies against multiple infectious diseases in serum and the airway mucosa using synthetic peptide-based linear epitope microarrays

doi: 10.1101/462689

Figure Lengend Snippet: The kinetics of maternal antibody decay were evaluated for five antigens on the peptide microarray chip. Changes in the levels of antigen-specific IgG in (A) serum and (B) airway mucosa, were analysed using loess curve fitting. For all antigens except SPNE, there was evidence of serum IgG decline in the first three months of life. In the airway mucosa similar changes in antigen specific IgG were observed. The dashed red lines indicate the 6-month age time point while the blue dashed line indicates the 12-month time point.

Article Snippet: Peptides were printed onto the epoxy slides using a non-contact inkjet microarray printer (Arrayjet, Scotland).

Techniques: Peptide Microarray