Journal: bioRxiv

Article Title: Extracellular Vesicle Antibody Microarray for Multiplexed Inner and Outer Protein Analysis

doi: 10.1101/2022.04.05.487234

Figure Lengend Snippet: EVPio: combinatorial profiling of extracellular vesicle inner and outer proteins. ( A) The EVPio assay is performed on a well gasket-covered glass slide inkjet-printed with 16 10×10 subarrays of antibodies, where each row targets a different EV surface protein. ( B) EVPio workflow: (1) capture antibodies are inkjet-printed on a functionalized slide (PolyAn 2D-Aldehyde), (2) the slide is washed and unoccupied sites are blocked, (3) the sample is incubated on the antibody spots and the EVs are captured based on their surface protein markers, (4) bound EVs are fixed on the slide, (5) the slide is AR-treated and (6) quenched to get rid of remaining reactive groups, (7) the slide is washed and incubated with up to three different oligo barcode-conjugated primary detection antibodies, (8) the slide is washed and incubated with complementary fluorescently-labeled oligonucleotide constructs (up to three colors), and (9) immobilized immunolabeled EVs are detected using a confocal microarray scanner. Microspots, antibodies, oligonucleotides and EVs are not to scale.

Article Snippet: The slides were then washed for 3 min in washing buffer, dried, and imaged using a confocal microarray scanner (InnoScan 1100 AL, Innopsys).

Techniques: Incubation, Labeling, Construct, Immunolabeling, Microarray