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…

Disclosures

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

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Cite This Article
A Snap Chip Technology for a Cross-Reactivity-Free Multiplexed Sandwich Immunoassay. J. Vis. Exp. (Pending Publication), e22137, doi: (2024).

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