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Multiplex Digital Immunoassays with Reduced Pre-partition Reaction
Weiyuan Lyu1, Jingwei Yi1, Qingsheng Guo1
1School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai 200030, China.
ACS Nano
|June 5, 2023
Summary
A novel bead-based SlipChip (bb-SlipChip) microfluidic device significantly reduces background noise in digital immunoassays. This innovation enhances protein analysis sensitivity and precision, improving quantification capabilities for multiplexed assays.
Area of Science:
- Biotechnology
- Microfluidics
- Assay Development
Background:
- Digital immunoassays offer ultrahigh sensitivity for protein analysis.
- Background signal from premixing limits precise quantification, especially in multiplexed assays.
Purpose of the Study:
- To present a bead-based SlipChip (bb-SlipChip) microfluidic device for digital immunoassay.
- To suppress background signal through a two-step sample loading mechanism.
Main Methods:
- A microfluidic device with massively parallel two-step sample loading was developed.
- Beads were encapsulated into microwells, followed by parallel delivery of fluorogenic substrate via a slipping process into 281,200 microwells (68 fL each).
Main Results:
- The bb-SlipChip suppressed background signal, enabling precise digital immunoassay.
- A duplex assay for IL-6 and IL-10 achieved limits of detection of 5.2 and 15.3 fg/mL, respectively.
- Performance was 2-3 times better than a commercial Simoa system.
Conclusions:
- The bb-SlipChip offers a robust and universal method for digital immunoassay.
- The technology can be extended for higher multiplexed detection and other microbead-based biomedical applications.
