Related Experiment Video
Updated: Jan 15, 2026
07:57
Quantification of Protein Interaction Network Dynamics using Multiplexed Co-Immunoprecipitation
Published on: August 21, 2019
9.1K
PiP-Plex: A Particle-in-Particle System for Multiplexed Quantification of Proteins Secreted by Single Cells
Félix Lussier1,2,3, Byeong-Ui Moon3, Mojra Janta-Polczynski3
1Biomedical Engineering Department, McGill University, 3775 Rue University, Montréal, QC, H3A 2B4, Canada.
Advanced Materials (Deerfield Beach, Fla.)
|October 7, 2025
Summary
This study introduces PiP-plex, a novel method for multiplex protein secretion analysis at the single-cell level. PiP-plex enables detailed characterization of cellular responses and potential in cell-based therapeutics.
Area of Science:
- Biotechnology
- Cell Biology
- Immunology
Background:
- Cell signaling proteins reveal cell phenotype but are difficult to measure multiplexed at the single-cell level.
- Existing methods lack the multiplexing capability for simultaneous detection of multiple secreted proteins from individual cells.
Purpose of the Study:
- To develop a novel platform, PiP-plex, for multiplexed analysis of protein secretion from single cells.
- To enable high-throughput single-cell secretion profiling using confocal microscopy.
Main Methods:
- Developed PiP-plex using fluorescence intensity barcoded microparticles (BMPs) co-entrapped with single cells in alginate hydrogel particles.
- Established a seven-plex fluorescent barcoding and sandwich immunoassay within PiPs.
- Validated PiP-plex performance against bulk immunoassays for sensitivity and accuracy.
Main Results:
- PiP-plex maintained high cellular viability (>90%) and allowed live cell retrieval.
- Achieved limits of detection from 0.8 pg/mL to 2 ng/mL for seven different proteins.
- Detected varying single-cell responses in THP-1 cells upon lipopolysaccharide exposure, identifying key secreted cytokines like MIP-1α and IL-17A.
Conclusions:
- PiP-plex is a robust and sensitive platform for multiplexed single-cell protein secretion analysis.
- The method reveals heterogeneity in cellular responses and has potential applications in characterizing cells for therapeutics, such as cancer immunotherapies.
