Rapid isolation of antigen-specific B-cells using droplet microfluidics

Area of Science:

• Biotechnology
• Immunology
• Microfluidics

Background:

• Traditional monoclonal antibody generation methods (hybridoma, display libraries) are inefficient and time-consuming.
• There is a need for rapid, high-throughput methods to identify antigen-specific antibody sequences from primary cells.

Purpose of the Study:

• To develop and validate a droplet microfluidics-based platform for rapid identification and verification of antigen-binding antibody sequences.
• To overcome limitations of existing antibody discovery techniques, such as fusion inefficiency and display bias.

Main Methods:

• Utilized droplet microfluidics combined with a bead-based binding assay for cell sorting.
• Characterized the system using hybridoma cells, achieving high-throughput sorting (100 Hz) and isolation of rare cells (0.1% input) with low false positive rates (<1%).
• Applied the platform to primary B-cells, recovering paired heavy- and light-chain antibody sequences via RT-PCR and generating single-chain variable fragment (scFv) antibodies.

Main Results:

• Successfully isolated rare hybridoma cells and applied the system to primary B-cells to identify target-binding antibody sequences.
• Generated functional scFv antibodies from sorted cells, with 12 out of 14 showing antigen-specific binding via ELISA.
• Demonstrated the platform's ability to screen B-cell repertoires rapidly and cost-effectively.

Conclusions:

• The developed platform significantly enhances the rate and range of discovering antigen-specific antibodies from living organisms.
• This microfluidic approach offers a rapid, low-cost solution for antibody discovery, adaptable for isolating cells based on various secreted products.
• The technology streamlines the process of identifying therapeutic and research antibodies from diverse biological sources.