Suspendable Hydrogel Nanovials for Massively Parallel Single-Cell Functional Analysis and Sorting.

Area of Science:

• Biotechnology
• Cell Biology
• Immunology

Background:

• Analyzing single-cell functional outputs, like secreted proteins, is crucial for advancing cell biology and therapies.
• Current methods face challenges due to rapid diffusion of secreted molecules and complex equipment needs.
• Compartmentalizing cells and capturing secretions is key for effective analysis.

Purpose of the Study:

• To develop and validate hydrogel-based nanovials for high-throughput, functional single-cell sorting.
• To demonstrate the nanovial platform's utility in selecting cells based on secreted protein production.
• To enable easier access to advanced cell screening technologies for researchers.

Main Methods:

• Fabrication of chemically functionalized, hydrogel-based microcontainers (nanovials) for cell attachment and secretion capture.
• Utilizing nanovials with commercial flow sorters for high-throughput screening of cells based on secreted products.
• Performing fluorescence immunoassays and single-cell RT-PCR on sorted cells.

Main Results:

• Successfully sorted Chinese hamster ovary (CHO) cells by IgG production, with sorted cells maintaining high secretion phenotype and increasing production rates by over 40%.
• Enriched hybridoma cells secreting specific antibodies from ~4% prevalence to over 90% in under an hour.
• Sorted and enriched antigen-specific antibody-secreting mouse B cells, demonstrating potential for antibody discovery workflows.

Conclusions:

• Nanovials provide a versatile and accessible platform for high-throughput functional single-cell sorting.
• The technology significantly improves the efficiency of selecting high-producing cells for therapeutic applications.
• This democratizes access to advanced cell screening, facilitating breakthroughs in cell and antibody therapies.