Journal: bioRxiv

Article Title: Salt-free fractionation of complex isomeric mixtures of glycosaminoglycan oligosaccharides compatible with ESI-MS and microarray analysis

doi: 10.1101/759993

Figure Lengend Snippet: Multi-dimensional separation workflow. Data shown are taken from this study to represent the value of each step. A.) SEC separation of the partially depolymerized Hp/HS, enoxaparin sodium; B.) chemo-selective reductive amination of one SEC fraction (dp8) with AEAB; C.) IPRP separation of AEAB-labeled oligosaccharide SEC fraction (dp8); IPRP fractions could be either analyzed by D.) online HILIC LC/MS (IPRP fraction RT 35 to 36 min shown); or E.) separated by offline HILIC with fractions collected (IPRP fraction RT 36.8 to 38 min shown); F.) microarray immobilization of HILIC fractions and protein binding affinity assay (IPRP and HILIC fraction identities shown in Table S1).

Article Snippet: 126 fractions after off-line HILIC separation were printed on NHS-ester activated glass slides (NEXTERION ® Slide H, Schott Inc.) using a Scienion sciFLEXARRAYER S3 non-contact microarray equipped with a Scienion PDC80 nozzle (Scienion Inc.).

Techniques: Labeling, Hydrophilic Interaction Liquid Chromatography, Liquid Chromatography with Mass Spectroscopy, Microarray, Protein Binding

Journal: bioRxiv

Article Title: Salt-free fractionation of complex isomeric mixtures of glycosaminoglycan oligosaccharides compatible with ESI-MS and microarray analysis

doi: 10.1101/759993

Figure Lengend Snippet: FGF2-binding assay on Hp/HS octasaccharide-AEAB microarray Each fraction is printed in replicates of 6 in a single block with droplet volume around 400 pL. Fractions from the multi-dimensional separation method are directly applicable to microarray study. Based on the fluorescent intensity of each spot, it indicates that each fraction has different binding affinities to FGF2. Fractions from No. 1 to No. 126 were eluents from the multi-dimensional separation method. Fractions No. 127 & 128 were unfractionated octasaccharide. Fraction No. 129 was synthetic octasaccharide as a positive control. An image of the microarray is in Supplementary Information Figure S11. Detailed information of each fraction is listed in Table S1.

Article Snippet: 126 fractions after off-line HILIC separation were printed on NHS-ester activated glass slides (NEXTERION ® Slide H, Schott Inc.) using a Scienion sciFLEXARRAYER S3 non-contact microarray equipped with a Scienion PDC80 nozzle (Scienion Inc.).

Techniques: Binding Assay, Microarray, Blocking Assay, Positive Control

Journal: bioRxiv

Article Title: Salt-free fractionation of complex isomeric mixtures of glycosaminoglycan oligosaccharides compatible with ESI-MS and microarray analysis

doi: 10.1101/759993

Figure Lengend Snippet: Correlation between IPRP retention time, HILIC retention time, and FGF2 binding response by microarray analysis at 1 μg/mL, represented by bubble size. A strong correlation between HILIC retention time and FGF2 binding is apparent, indicating that FGF2 is binding ligands that are more highly sulfated. No correlation between IPRP retention time and FGF2 binding is apparent.

Article Snippet: 126 fractions after off-line HILIC separation were printed on NHS-ester activated glass slides (NEXTERION ® Slide H, Schott Inc.) using a Scienion sciFLEXARRAYER S3 non-contact microarray equipped with a Scienion PDC80 nozzle (Scienion Inc.).

Techniques: Hydrophilic Interaction Liquid Chromatography, Binding Assay, Microarray

Journal: Analytical Chemistry

Article Title: A Novel Quantitative Multi-Component Serological Assay for SARS-CoV-2 Vaccine Evaluation

doi: 10.1021/acs.analchem.1c05264

Figure Lengend Snippet: LLOD of the microarray tests. Upper panel: Calculated MFI values (after blank subtraction) for (left to right) IgG, IgA, and IgM signals of the commercial, pre-pandemic, naïve samples (eight independent experiments) on S2P (orange), RBD (blue), and NC (green). The average (AV), standard deviation (STD), and lower limit of detection (LLOD) for each antibody isotope for each antigen are indicated on the graph (bold black lines and dashed colored lines, respectively), and the values are presented in the lower panel. LLOD values are indicated in bold. No LLOD was determined for IgM-NC, since no acceptance range was determined for this test (due to a high CV value observed during acceptance range determination, Figure S2 ).

Article Snippet: SARS-CoV-2 antigens including S2P, RBD, and NC (1 mg/mL) were spotted separately, as single 300 pL drops, in 18 repeats on 16-pad (16 sub-arrays) nitrocellulose-coated slides (Grace Bio Labs, GBL, Bend, OR) using a non-contact Piezo dispensing microarray spotter (Scienion Inc., Berlin, Germany).

Techniques: Microarray, Standard Deviation

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