Barcoded sequencing workflow for high throughput digitization of hybridoma antibody variable domain sequences

Area of Science:

• Biotechnology
• Immunology
• Genomics

Background:

• Hybridoma technology, developed in 1975, revolutionized antibody discovery by enabling functional screening and long-term storage of antibody-producing B cells.
• Advancements in robotics and automation have increased the identification of antigen-specific hybridoma clones, leading to challenges in storage capacity and antibody sequencing throughput.

Purpose of the Study:

• To develop a high-throughput antibody sequencing workflow to address limitations in physical storage and sequencing capacity.
• To provide bioinformatics tools for robust and unambiguous antibody sequence reporting.

Main Methods:

• Implementation of a barcoded sequencing workflow utilizing next-generation sequencing (NGS).
• Development of accompanying bioinformatics tools for data analysis and sequence determination.
• Validation of the workflow using Sanger sequencing as a control.

Main Results:

• The barcoded sequencing workflow significantly expands conventional sequencing capacity.
• The workflow robustly reports unambiguous antibody sequences, confirmed by Sanger sequencing.
• Digitization of antibody sequences is achieved efficiently, complementing recombinant DNA technology.

Conclusions:

• The barcoded sequencing workflow offers an effective solution to storage and sequencing capacity limitations in antibody discovery.
• This approach enables high-throughput digitization of antibody sequences, supporting future research and development.
• The method enhances the scalability and efficiency of monoclonal antibody discovery pipelines.