Journal: Journal of Immunological Methods

Article Title: Agrodiag PorCoV: A multiplex immunoassay for the differential diagnosis of porcine enteric coronaviruses

doi: 10.1016/j.jim.2020.112808

Figure Lengend Snippet: Overview of the design, fabrication and use of the AgroDiag porcine enteric coronavirus (PEC) multiplex planar immunoassay. (A) Manufacturing steps include functionalization of the wells of a microplate with a dextran layer before printing of the protein array in triplicate. (B) The assay protocol and working principle is similar to the solid-phase standard indirect ELISA. After the three typical incubation steps (serum sample, conjugate, and substrate), followed by microplate reading. The reaction was visualized as brown spots which intensity correlated with antibody levels to specific viral antigen target in the array. Four results are obtainable depending on incubation with a negative control, or samples from PDCoV, PEDV, TGEV inoculation groups. Scale bar corresponds to 1 mm. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Supplementary Fig. S1 Supplementary Fig. S2 AgroDiag PEC multiplex immunoassay raw data obtained with the microplate reader (CLAIR Sensovation®) on serum samples, tested in duplicate, collected at day post-inoculation −7, 0, 7, 14, 21, 28, 35, and 42 from pigs ( n = 12, per group) experimentally inoculated with: (A) culture media (NEG); (B) porcine deltacoronavirus (PDCoV); (C) porcine epidemic diarrhea virus (PEDV); (D) transmissible gastroenteritis virus (TGEV) Purdue; (E) TGEV Miller.

Techniques: Multiplex Assay, Protein Array, Indirect ELISA, Incubation, Negative Control

Journal: Biosensors & Bioelectronics

Article Title: Array-based analysis of SARS-CoV-2, other coronaviruses, and influenza antibodies in convalescent COVID-19 patients

doi: 10.1016/j.bios.2020.112643

Figure Lengend Snippet: Representative AIR array images of (A) 5% FBS; (B) 10% PNHS; (C) a negative single-donor sample, and (D) one convalescent serum sample. Strong responses to SARS-CoV-2 antigens are readily observed in (D), but not in (A), (B), or (C), while responses to circulating coronaviruses HKU, OC43, and 229E are observed in (B), (C), and (D). In each case, samples were diluted 1:20 in Adarza diluent, and incubated with the arrays overnight at 4 °C. See for key to the array. All arrays in this figure were imaged at an exposure of 100 ms.

Article Snippet: Each spot consists of approximately 300 pixels when imaged by the CCD in an AIR chip reader (Adarza BioSystems, Inc.), with each pixel representing a discrete interrogation of a unique probe surface region.

Techniques: Incubation

Journal: Biosensors & Bioelectronics

Article Title: Array-based analysis of SARS-CoV-2, other coronaviruses, and influenza antibodies in convalescent COVID-19 patients

doi: 10.1016/j.bios.2020.112643

Figure Lengend Snippet: AIR results from convalescent COVID-19-positive subjects. Empty cells indicate unreadable spots. (A) Each sample was diluted 1:5 in Adarza diluent, and incubated with the array overnight at 4 °C. (B) Selected antigen results for samples run at 1:20 dilution, and incubated with the array overnight at 4 °C. All values reported are in Ångstroms of build relative to an FBS control.

Article Snippet: Each spot consists of approximately 300 pixels when imaged by the CCD in an AIR chip reader (Adarza BioSystems, Inc.), with each pixel representing a discrete interrogation of a unique probe surface region.

Techniques: Incubation

Journal: PLoS ONE

Article Title: Targeted Deposition of Antibodies on a Multiplex CMOS Microarray and Optimization of a Sensitive Immunoassay Using Electrochemical Detection

doi: 10.1371/journal.pone.0009781

Figure Lengend Snippet: Reagents are loaded in a 96-well plate, and a programming script is used to control their flow into a single chamber or into four chambers containing an ElectraSense microarray. Current or voltage can be applied to single or groups of electrodes for Ppy deposition, and current can be read from a single electrode or groups of electrodes for ECD.

Article Snippet: Hybridization to probes can be detected using cyanine (Cy) dyes and fluorescent scanners or, alternatively, using horseradish peroxidase (HRP) and enzyme-enhanced electrochemical detection (ECD) on CombiMatrix's microarray readers.

Techniques: Microarray

Journal: PLoS ONE

Article Title: Targeted Deposition of Antibodies on a Multiplex CMOS Microarray and Optimization of a Sensitive Immunoassay Using Electrochemical Detection

doi: 10.1371/journal.pone.0009781

Figure Lengend Snippet: A) Photomicrograph of an ElectraSense microarray under epi illumination and showing the deposition of polypyrrole with adsorbed murine IgG on a 5×5 set of electrodes. B) Fluorescence scanned image of the microarray treated with Cy5-labeled goat anti mouse IgG and showing the presence of murine Ab on the electrodes.

Article Snippet: Hybridization to probes can be detected using cyanine (Cy) dyes and fluorescent scanners or, alternatively, using horseradish peroxidase (HRP) and enzyme-enhanced electrochemical detection (ECD) on CombiMatrix's microarray readers.

Techniques: Microarray, Fluorescence, Labeling

Journal: PLoS ONE

Article Title: Targeted Deposition of Antibodies on a Multiplex CMOS Microarray and Optimization of a Sensitive Immunoassay Using Electrochemical Detection

doi: 10.1371/journal.pone.0009781

Figure Lengend Snippet: A ) Three concentrations (0.003, 0.01, and 0.03 pg/ml) were incubated on a microarray with either anti-SEB MAb (•-•) or casein (▪-▪) adsorbed onto Ppy that was deposited at 40 nA for 1 s. SEB binding was detected using biotinylated rabbit anti-SEB Ab and SA-HRP. Background (0 pg/ml SEB) was 1147±283. B ) Different concentrations of SEB were incubated on a microtiter plate with anti-SEB MAb adsorbed onto the surface of each well. SEB binding was detected using biotinylated rabbit anti-SEB Ab and SA-HRP. Background (0 pg/ml SEB) was OD 450–570 2.939±.026.

Article Snippet: Hybridization to probes can be detected using cyanine (Cy) dyes and fluorescent scanners or, alternatively, using horseradish peroxidase (HRP) and enzyme-enhanced electrochemical detection (ECD) on CombiMatrix's microarray readers.

Techniques: Incubation, Microarray, Binding Assay