A microwell array platform to print and measure biomolecules produced by single cells

Area of Science:

• Biotechnology
• Cell Biology
• Immunology

Background:

• Single-cell analysis is crucial for understanding cellular heterogeneity and function.
• Existing methods for monitoring secreted molecules and isolating single cells are often complex or lack precision.
• Identifying and isolating specific antibody-producing cells is vital for therapeutic development.

Purpose of the Study:

• To develop and validate a combined method for monitoring single-cell molecule secretion and subsequent cell isolation.
• To enable precise correlation between secreted molecules and their producing single cells.
• To demonstrate the utility of the method in antibody-producing cell lines.

Main Methods:

• A self-sorting microwell chip integrated with an activated membrane for molecule collection.
• Diffusion-based printing of secreted molecules onto the membrane in discrete spots.
• Correlation of molecule spot locations with microwell and cell identity.
• Application to hybridoma (VU1D9) and engineered CHO cell lines.

Main Results:

• The method successfully monitored and correlated molecule secretion with individual cells.
• VU1D9 hybridoma cells secreted EpCAM antibody at 4.6 ± 5.6 pg/24h.
• Engineered CHO cells produced Herceptin antibody at 6.5 ± 8.2 pg/24h.
• Demonstrated feasibility for analyzing antibody production in different cell types.

Conclusions:

• The described method offers a powerful tool for single-cell analysis of secreted molecules.
• It facilitates the identification and isolation of high-producing cells for research and therapeutic applications.
• This technique advances single-cell biotechnology and antibody discovery platforms.