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Updated: May 7, 2026
A Microfluidic Chip for the Versatile Chemical Analysis of Single Cells
Published on: October 15, 2013
Hemin-DNA Switch-Assisted Microfluidic Chemiluminescence Immunoimaging for Rapid and Quantitative Screening of
Wencheng Xiao1, Hang Ao1, Nilin Wu1
1State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing 210023, China.
Insights
A new microfluidic chemiluminescence immunoimaging method rapidly screens single hybridoma cells for high antibody production. This sensitive technique enables efficient monoclonal antibody (mAb) screening and production.
Area of Science:
- Biotechnology
- Analytical Chemistry
- Cell Biology
Background:
- Sensitive detection of single-cell secretions is vital for monoclonal antibody (mAb) production.
- Current screening methods can be time-consuming and lack sensitivity for high-yield hybridoma cell selection.
Purpose of the Study:
- To develop a highly sensitive and rapid microfluidic chemiluminescence immunoimaging (μCLII) method for screening high-yield specific hybridoma cells (HSHCs).
- To enable quantitative, single-cell detection of secreted antibodies for efficient mAb production.
Main Methods:
- Utilized a lantern-shaped microchamber array chip for cell capture and isolation.
- Employed a proximity recognition amplification strategy involving DNA hybridization and rolling circle amplification (RCA).
- Integrated hemin-DNA enzymes and a luminol-H2O2 system for chemiluminescence (CL) signal generation.
Main Results:
- Achieved a detection limit of approximately 60 antibodies per chamber.
- Enabled precise sorting of HSHCs within 70 minutes.
- Demonstrated high specificity and yield ( >80 fg) for antiproprotein convertase subtilisin/kexin type 9 mAb (PCSK9-mAb) producing cells.
Conclusions:
- The μCLII method offers a sensitive and rapid approach for single-cell analysis and HSHC screening.
- This technique significantly enhances efficiency in monoclonal antibody production.
- The platform shows great potential for various single-cell secretion analysis applications.
Abstract:
Highly sensitive detection of cell secretions at the single-cell level is critical for efficient cell screening and monoclonal antibody (mAb) production. Herein, we present a highly sensitive microfluidic chemiluminescence immunoimaging (μCLII) method for rapid and quantitative screening of high-yield specific hybridoma cells (HSHCs) by detecting single-cell secreted antibodies on a rationally designed lantern-shaped microchamber array chip. This method can effectively capture and isolate single hybridoma cells to in situ detect their secreted antibody by visual CL imaging with a proximity recognition amplification strategy. The proposed strategy can be performed by antibody-triggered proximity hybridization to release a DNA sequence, which initiates a rolling circle amplification reaction to turn on abundant hemin-DNA enzymes for generating strong CL emission of the imidazole-enhanced luminol-H2O2 system. This method shows a detection limit of ∼60 antibodies per chamber, and thus enables a precise sorting of HSHCs within 70 min. Using antiproprotein convertase subtilisin/kexin type 9 mAb (PCSK9-mAb) as a model, all of the cells selected by the μCLII method show high-producing hybridomas with PCSK9-mAb yield more than 80 fg during the screening process, indicating a promising potential of the proposed method for single-cell analysis and mAbs screening.
