Journal: Nature chemistry

Article Title: Synthetic O-acetylated sialosides facilitate functional receptor identification for human respiratory viruses.

doi: 10.1038/s41557-021-00655-9

Figure Lengend Snippet: Fig. 4 | Glycan microarray studies reveal binding patterns associated with host specificities. a, Column graphs of viral receptor selectivities. See Supplementary Fig. 9 for the chemical structures of the compounds. Additional results with different concentrations of proteins are presented in Supplementary Fig. 12. Columns show the background-subtracted average relative fluorescence unit (RFU) values of four replicates. Error bars indicate the s.d. of the RFUs. b, Heat-map presentations of viral receptor selectivities. In the heat map, the signal intensities are normalized with the highest value in each protein defined as 1.0 and shown with a colour gradient. Concentrations of Fc-tagged proteins presented are: BCoV S1A 0.3 μg ml–1, OC43 S1A 0.3 μg ml–1, HKU1 S1A 30 μg ml–1, ICV HEF 0.3 μg ml–1, IDV HEF 0.3 μg ml–1, porcine torovirus (PToV) HE 3 μg ml–1, BToV HE 3 μg ml–1, BCoV HE 3 μg ml–1, ECoV HE 3 μg ml–1, RbCoV HE 3 μg ml–1, CRCoV HE 3 μg ml–1, MHV-S HE 3 μg ml–1, ECoV S1A 3 μg ml–1, RbCoV S1A 3 μg ml–1 and CRCoV S1A 10 μg ml–1. Representative surface dissociation constants (Kd,surf) were obtained for the binding of HKU1 S1A with compounds 3g (9-O-Ac, 60 nM) and 3d (7,9-di-O-Ac, 85 nM). See supplementary Fig. 13 for binding curves. Sialoforms 1b and 2b (4,7-di-O-Ac Neu5Ac) were not included in the library because they have not been documented to be naturally occurring.

Article Snippet: The biotinylated compounds were printed on streptavidin-coated glass slides (SuperStreptavidin Microarray Substrate Slides, ArrayIt Inc) using a Scienion sciFLEXARRAYER S3 non-contact microarray equipped with a Scienion PDC80 nozzle (Scienion Inc).

Techniques: Glycoproteomics, Microarray, Binding Assay, Fluorescence

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