Journal: Lab on a chip
Article Title: High-resolution low-cost LCD 3D printing for microfluidics and organ-on-a-chip devices.
doi: 10.1039/d3lc01125a
Figure Lengend Snippet: Fig. 5 LCD 3D printed microfluidic ELISA chip. (a) ELISA-on-a-chip designed for LCD 3D printing with structurally encoded sequential delivery of reagents to autonomously perform an assay and coupled with a built-in chip-to-assay connection. (b) ELISA workflow showing the sandwich immunoassay designed for the detection of IFN-γ by sequentially delivering assay reagents and wash buffer to a nitrocellulose membrane pre- spotted with anti-human IFN-γ capture antibody; (1) sample containing IFN-γ, (2) biotinylated anti-human IFN-γ detection antibody, (3) streptavidin-conjugated enzyme pHRP, and (4) enzyme substrate in the presence of hydrogen peroxide to generate the colorimetric readout. (c) Autonomous CC workflow for on-chip reagent aliquoting by metering the correct reagent volumes and drainage of the excess, followed by (d) MCR-based sequential delivery of assay reagents with wash steps in between. Arrows show the direction of flow. Scale bar = 5 mm. (e) Binding curve of the on-chip assay for the detection of IFN-γ with a limit of detection of 12 pg mL−1 (CV: 6.8%) across triplicate chips for each tested concentration point; line shows a 4-point logistic fit.
Article Snippet: Purified mouse monoclonal IgG antihuman interferon-γ capture antibody (Cat. #MAB2852, lot #FIO1022021, R&D Systems, Minneapolis, Minnesota, United States), biotinylated affinity purified goat IgG anti-human interferon-γ detection antibody (Cat. #BAF285, lot #ZX2721071, R&D Systems, Minneapolis, Minnesota, United States), recombinant human interferon-γ protein (Cat. #285- IF, lot #RAX2422031, R&D Systems, Minneapolis, Minnesota, United States), Pierce streptavidin poly-horseradish peroxidase (pHRP) (Cat. #21140, lot #XJ360080, Thermo Fisher Scientific, Waltham, Massachusetts, United States), SIGMAFAST 3,3′-diaminobenzidine tablets (Cat. #D4293, lot #SLCG5357, Sigma-Aldrich, Oakville, Ontario, Canada), bovine serum albumin (BSA) (Cat. #001-000-162, lot #162191, Jackson ImmunoResearch Labs, West Grove, Pennsylvania, United States), BSA-biotin (Cat. #A8549, Sigma-Aldrich, Oakville, Ontario, Canada), Tween 20 (Cat. #P7949, lot #SLBX0835, Sigma-Aldrich, Oakville, Ontario, Canada).
Techniques: Enzyme-linked Immunosorbent Assay, Membrane, Binding Assay, Concentration Assay
