A Snap Chip Technology for a Cross-Reactivity-Free Multiplexed Sandwich Immunoassay
Published: April 30, 2024
Abstract
Source: Li, H. et al., Snap Chip for Cross-reactivity-free and Spotter-free Multiplexed Sandwich Immunoassays. J. Vis. Exp. (2017)
The video showcases the implementation of snap chip technology for cross-reactivity-free multiplexed sandwich immunoassays. The simple action of snapping two microarray slides, each containing different reagents, ensures reagent transfer without cross-contamination and helps to detect multiple proteins using sandwich immunoassay.
Protocol
All procedures involving sample collection have been performed in accordance with the institute's IRB guidelines. 1. Fabrication and storage of snap chips Single transfer method (Figure 1a) Spot cAb solutions containing 400 µg/mL antibodies and 20% glycerol in phosphate-buffered saline (PBS) onto a nitrocellulose (or a functionalized glass) assay slide with an inkjet microarray spotter at a relative humidity of 60% (1.2 nL…
Representative Results
Figure 1. Schematic of assay procedure comparing (a) single and (b) double transfer methods. Figure 2. Schematic showing differences between single- and double-transfer. (a) mirroring of the transfer reagents amplifies the angular misalign…
Divulgations
The authors have nothing to disclose.
Materials
| Phosphate buffered saline tablet | Fisher Scientific | 5246501EA | |
| Streptavidin-conjugated Cy5 | Rockland | s000-06 | |
| Tween-20 | Sigma-Aldrich | p1379 | |
| Bovine serum albumin | Jackson ImmunoResearch Laboratories, Inc | 001-000-162 | |
| Glycerol | Sigma-Aldrich | G5516 | |
| Blocking solution: BSA-free StabilGuard Choice Microarray Stabilizer | SurModics, Inc | SG02 | |
| Nitrocellulose coated slides | Grace Bio-Laboratories, Inc | 305116 | |
| Aminosilane coated slides | Schott North America | 1064875 | |
| Snap Device | Parallex BioAssays Inc. | PBA-SD01 | |
| Inkjet microarray spotter | GeSiM | Nanoplotter 2.0 | |
| Slide module gasket | Grace Bio-Laboratories, Inc | 204862 | |
| Humidity Stabilization Beads | Parallex BioAssays Inc. | PBA-HU60 | |
| Array-Pro Analyzer software | Media Cybernetics | Version 4.5 | |
| Fluorescence microarray scanner | Agilent | SureScan Microarray Scanner | |
| Biostatistics software | GraphPad Software | GraphPad Prism 6 | |
| Endoglin capture antibody | R&D Systems | MAB10972 | |
| Endoglin protein | R&D Systems | 1097-EN | |
| Endoglin detection antibody | R&D Systems | BAF1097 | |
| IL-6a (see Table 1) | R&D Systems | ||
| IL-6b (see Table 1) | Invitrogen |
Tags
Citer Cet Article
A Snap Chip Technology for a Cross-Reactivity-Free Multiplexed Sandwich Immunoassay. J. Vis. Exp. (Pending Publication), e22137, doi: (2024).